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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30General SciencesA Study on Awareness of Maternal and Child Health Care Schemes under National Health Mission in Majuli, Assam
English0105Bornali DuttaEnglish Manash Pratim BarmanEnglishAim: In this paper an attempt has been made to accesses the level of awareness on maternal and child health care schemes among women of reproductive age and to examine the association between different socio-demographic factors and awareness level of the respondents.
Methodology: Data are collected using cluster sampling method. Statistical significance is tested using Chi-square test for independence of attributes and significance level is considered at p- value < 0.05.
Results: Caste of the respondents, order of birth of the children and road communication of the study area has significant association with the awareness level.
Conclusion: Efforts are needed to increase awareness level of the reproductive women in the rural areas like Majuli regarding various free entitlements of NRHM
EnglishNRHM, Majuli, JSSK, cluster sampling, Reproductive womenIntroduction:
In many parts of India, especially in rural areas, the main role assigned to women is that of child bearer. Thus, pregnancy is one of the most important events in the life of Indian women. Pregnancy –related complications are among the leading causes of death and disability for women age 15-49 years in the developing countries including India [4]. Most of the maternal deaths are preventable with good antenatal care, timely identification and referral of pregnant women with complications of pregnancy and timely provision of emergency obstetric care [1]. Therefore, India government introduced the national population policy (NPP-2000) with defined goal to increase institutional delivery by 80.0%, safe delivery by 100% and reducing Maternal Mortality Rate (MMR) by 100 per 1,00,000 live births[1]. The crucial importance of maternal health is underscored by the 5th goal of the United Nations Millennium Development Goals (MDG) which is aiming at improving maternal health. Millennium Development Goal-5 aims to reduced the Maternal Mortality Rate (MMR) 109 per 1, 00,000 live births by 2015[4]. India has made considerable progress towards the reduction of Maternal Mortality Ratio (MMR) and Infant Mortality Rate (IMR), but the current pace of decline is not sufficient. According to Millennium Development Goal - 2015 report, Maternal Mortality Rate (MMR) in India is 140 per 1, 00,000 live births and the state of Assam shows worst performance in reducing Maternal Mortality Rate (MMR). Still Maternal Mortality Rate (MMR) is highest in Assam at 300 per 1, 00,000 live births [4]. To achieve the Millennium Development Goals (MDG), Ministry of Health and family Welfare, Govt of India launched a nationwide initiative - National Rural Health Mission (NRHM) in 2005 and National Urban Health Mission (NUHM) in 2013 and which is popularly known as National Health Mission (NHM). Under National Health Mission (NHM) Govt. of India as well as our state government Assam introduced various schemes related to maternal and child health care. The central government implemented Janani Shishu Suraksha Karyakram (JSSK) and Janani Suraksha Yojna (JSY) schemes and our state government Assam also implemented various schemes: Majoni, Mamoni, Mamata in order to improve maternal and child health care. However, the key for the successful implementations of these schemes is the level of awareness among the beneficiaries. Awareness regarding various maternal and child health care schemes among the reproductive women is not much studied in Assam. With this background the present study is conducted to assess the level of awareness on maternal and child health care schemes among the women in the reproductive age in Kamalabari block of Majuli district, Assam.
Description of the Study Area:
The researcher collected data from Kamalabari block under Majuli district. Majuli is the world's largest river island and it is a newly announced district by the Assam government.
It is a flood affected area. Due to flood and erosion, the transport and communication system of Majuli is not good. Cultivation is the main occupation of the villagers. Handloom is a major occupation among the women of the villages. The island Majuli has been the hub of Assamese neo-vaishnavite culture, initiated around 15th century by the revered Assamese saint Srimanta Sankardev and his disciple Madhavdev. Many satras or monasteries constructed by the saint still survive and represent the colorful Assamese culture. The Samuguri Satra is famous for the mask making in India.
Methodology:
The design of the study is cross sectional and it is based on household investigation. The study is conducted in the Kamalabari block of Majuli District of Assam during 2016. Reproductive women residing in the study area during the study period constituted the study subjects. The researcher set inclusion and exclusion criteria for the study subjects to be included in the sample. The inclusion criteria are: (i) women in the reproductive age group (ii) the women who have children in the last three years. The exclusion criterion is: (i) the women who are not present in the study area during the study time. In the present study the sample size is 216; which is reasonable as WHO (World Health Organization, 2005) developed a 30 by 7 cluster sampling method for conducting similar type of study [6]. In this study, the Kamalabari block of Majuli district is selected randomly. In the selected block, 12 villages are selected by using probability proportional to size sampling method. Again from each of 12 selected villages, 18 women satisfying inclusion criteria are selected using systematic sampling method in the survey. During the survey, local ASHA helped in locating the houses. In the survey, data on awareness about different maternal and child health care schemes of NRHM are collected. The response is recorded into three categories viz, spontaneously answered, answered after probing and no awareness. If the respondent spontaneously answered about the facility; it is put in the spontaneously answered category and if the respondents answered any facility after recall by the investigator then it is put into answered after probing category and the respondent who are unable to answer by probing and spontaneously; it is put into no awareness about the facility.
Socio-economic status of the study subjects are collected by using instrument developed by Parasuram S.et al [3]. All the data are analyzed in SPSS, version 17.0 software. Statistical significance are tested using Chi-square test for independence of attribute and significance level is considered at p- value Englishhttp://ijcrr.com/abstract.php?article_id=2284http://ijcrr.com/article_html.php?did=22841. Begum Rasida (2014): "Maternal Health care services in India with special reference to Suraksha Yojna in Assam"- A peer-reviewed indexed International Journal of Humanities and Social Science, ISSN: 2278-5264(Online), volume-III, Issue-II
Chatterjee S.et al (2015): "Awareness about Janani Shishu Surakha Karyakram (JSSK) among pregnant mothers - a community based study in a rural area of West Bengal, India" IOSR Journal of Dental and Medical Sciences, p- ISSN: 2279-0861. Volume14, Issue 9.
Parasuram S.et al (1999): "Role of Women's Education in India: Evidence from National Family Health Survey", Himalaya Publishing House, Mumbai.
Millennium Development Goals India Country Report 2015, social statistics division ministry of statistics and programme implementation government of India.
International institute for population sciences and macro international, National Family health survey (NFHS-3), 2005-06 India, Mumbai.
World Health Organization (2005).Immunization coverage cluster survey-Reference manual. Geneva, Switzerland: Department of Immunization, vaccines and Biological, Family and community Health. World Health Organization.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30General SciencesInnovative Teaching Methods in Physical Education for Better Learning
English0611Ramananda NingthoujamEnglish Bidhyapati NongthombamEnglish Moirangthem SunderchandEnglishIntroduction: Physical educators have been trying to instil values of fitness for life in the mind of the children. The NASPE (2003) and Institute of Medicine’s (2013) recommend that children should obtain a minimum of 60 minutes of moderate to vigorous physical activity (MVPA) in a day.
Aim: This article will highlight the latest teaching strategies, age appropriate equipments, and health optimizing physical education curriculum used in the field of PE program for better teaching and learning process.
Methods: New physical education activities, equipments, physical education curriculum in the field of physical education obtained through Google search, with the keywords physical education curriculum, physical education equipments, age appropriate physical education program, assessment in physical education, Strategies in physical education, and curriculum guidelines for physical education; as well the experience by researchers were expressed in this article.
Discussion: A physical educator has a great role to play at the grass root level to groom a child into making a potential and highly skilled professional. Olympians are not born, but they are made through their lifetime participation in research based PE program from school.
Conclusions: A physical educator has a great role to play in the grass root level to groom a child into making a potential and highly skilled professional. New curriculum, age appropriate equipments, teaching techniques, etc should be introduce in all the schools to make the child engage in physical activity for at least 60 minutes per day. However, the use of innovative equipments, strategies and curriculum in PE classes must be research-based and need for the children.
EnglishPhysical Education, Curriculum, Equipments, School ChildrenIntroduction
Physical Education (PE) programme has been creating, implementing, and evaluating in promoting lifelong wellness among the school children. Students learn how to make healthy food choices, reduce time playing video games and watching television (Sallis, J. F., and McKenzie, T. L., 1991) 1. Previously, the PE program has been removed from the school curriculum as schools everywhere strive to improve the academic performance of their students, many have cut physical education and recess periods to leave more time for sedentary classroom instruction (Stewart G. et al, 2009)2. Over the years, researchers and educators have revised their PE curricula, equipments and training programs to meet the six NASPE (National Association of Sport and Physical Education) guidelines. In India, Central Board of School Education (CBSE) has made PE an elective subject in school curriculum. The idea of including PE class in school is to make the students healthy and teach them healthy life style. The aim of the study is to highlight the latest strategies, age appropriate equipments, new curriculum, used in the field of PE program for better learning and developing health among young children through physical activity. For this purpose, new physical education activities, equipments, physical education curriculum in the field of physical education obtained through Google search, with the keywords physical education curriculum, physical education equipments, age appropriate physical education program, assessment in physical education, Strategies in physical education, and curriculum guidelines for physical education; as well the experience by researchers were expressed in this article.
Innovative program in Physical Education
Till date the Physical education program in some schools in India have not really changed its methodology, wherein children play a particular sport by sharing one ball in a huge group. And the drawback is that not every child gets the opportunity to play. With the introduction of new curriculum, age appropriate equipments, teaching techniques, etc the physical education class have become more interesting, attractive and more valuable for the students. Researchers and educators are trying to revise their PE curricula and training programs to meet the six NASPE (National Association of Sport and Physical Education) standard guidelines: 1) demonstrates competency in motor skills and movement patterns needed to perform a variety of physical activities, 2) demonstrates understanding of movement concepts, principles, strategies, and tactics as they apply to the learning and performance of physical activities, 3) participates regularly in physical activity, 4) achieves and maintains a health-enhancing level of physical fitness, 5) exhibits responsible personal and social behaviour that respects self and others in physical activity settings, and 6) values physical activity for health, enjoyment, challenge, self-expression, and/or social interaction.
SPARK (Sports, Play, and Active Recreation for Kids), a pioneer organisation in United States, has been contributing in the systemic reform of physical education (PE) since 1989. In addition to the six guidelines of NASPE, SPARK supports Healthy People Goals (HPG) 2010: a) PE classes make every effort to engage students in moderate to vigorous physical activity (MVPA) at least 50% of their class time without sacrificing academic achievement or student enjoyment of PE, b) SPARK focus on positive health outcomes of students. These include physical fitness and sports skills, c) SPARK programs include strategies that promote behaviour and environmental change techniques, and d) SPARK also follows ASAP (As Soon As Possible) activity to avoid the lengthy instruction in Physical education class (SPARK)3.
In India, many organisations such as LEAPSTART (FITKIDS), EDUSPORTS, KOOH SPORTS, SPORTS MENTOR, Physical Education Foundation of India (PEFI), Youth Affairs and Sports (YAS), National Association of Physical Education and Sports Science (NAPESS), and all the physical educators and health educators are also taking initiatives to develop physical education programme to meet the standard guidelines. Recently, Sports Authority of India (SAI) in collaboration Global Trust has launched a new scheme in India called the School Sports Promotion Foundation (SSPF) in 5 sports discipline viz. Athletics, Football, Volleyball, Basketball, and Cricket for talent identification in school children. Its main objective is to provide sports education in children from the grass root level. Sports education includes basics movement’s skills, locomotion, basic skills of the games, rules and regulations of the games, nutritional knowledge, team cohesion, etc (indiatoday)4.
Teaching Strategies
New teaching strategies like AS SOON AS POSSIBLE (ASAP) active games were introduced in PE class to keep the students active as soon as possible, which is one of the physical educator’s objectives. In many traditional PE classes, students arrive at the activity area only to stand or sit and listen to lengthy instructions (SPARK)5. This wastes valuable activity time and should, therefore, be done sparingly. This section includes a variety of enjoyable activities to begin activity immediately. ASAP Activities use little or no equipment, are fun and challenging, and promote health related fitness. They quickly involve all students in moderate-to-vigorous physical activity (MVPA) with few instructions or rules to slow them down. It reduces wasting of time in giving lengthy instruction in the class. Playground Expectation cards, Skills cards, Task cards, etc were also used on the playfield to reduced lengthy instructions.
Another strategy used in PE class is 80/20 RULES, which means that the physical education teacher tries to keep the students active 80% of class duration. Music is also used in PE classes to make more fun activities which interest the students and are also used as start and stop signals.
Limited space activities like BINGO GAMES are introduced which allowed the PE class to run in limited space since space constraint is one of the most common issues in today`s school environment. Whether it be inclement weather (rainy day, smog warnings, extreme heat, etc.) or the usual activity area is unavailable (assembly in the gym, a book fair in the multi-purpose room, the blacktop is getting re-paved, etc.), every so often, the only choice of space for PE is a classroom or other small area. To involve the parents` in PE activities HOME – PLAY games were introduced in PE class which keep the child active at home, and during vacation. It is a take-home page with interesting facts on 1 side, and fun challenges on the other.
Moderate to Vigorous Physical Activity (MVPA) is also introduced during the PE classes to keep the students ready for the next challenges. For example, Chasing and fleeing skills are used in many of the most popular games played by children. By changing the locomotor skill, the method of tagging, the task students perform to re-enter the game after being tagged, and/or the ways in which they may seek safety, you can create enough tag games to last a lifetime. Chasing and fleeing games are usually short in duration (5-10 minutes), and use little or no equipment. The games in this unit provide opportunities for students to develop chasing, fleeing and spatial awareness skills while promoting health-related fitness in enjoyable ways. The activities are designed to include and challenge all students’ physical skills, while maintaining enjoyable, health-promoting, moderate-to-vigorous physical activity during class.
Latest Equipments:
The equipment is one of the backbones of PE classes. In PE classes, it should be age-appropriate and safe to use for the child. Bevans K.B. et al., (2010)6 suggested that age-appropriate equipment increases pain free practice, increases students’ chances for active participation in the class and maximize the amount of time devoted to PE during which children are physically active. Children get maximum activity when every child has a piece of equipment and does not have to wait a turn (Judith E. R et al., 2010)7.
For early childhood, equipments like scarf and balloon (Figures 1a, 1b) are used in learning catching and throwing lessons. In K-2 grade students, bean bags, fluff ball (Figures1c, 1d) are used which falls quickly to gravity and the movement becomes fast. A foam ball (Figures 1e, 1f) is another equipment introduced to teach kicking and trapping in K-2 and 3-6 children. Fun activities like parachute and manipulative games (Figures 1g, 1h) which teach them team work, and develop their upper body strength were introduced to them. All the objects are safe to play and doesn`t hurt the child.
Latest Curriculum:
The National Association for Sport and Physical Education (NASPE) is now endorsing the concept of Comprehensive School Physical Activity Programs (CSPAP) that are designed to increase daily levels of physical activity for all school-age children and youth (Figure 2 ).This new CSPAP curriculum model called Health Optimizing Physical Education (HOPE) that can be used to help Primary to High School (P-12) students to acquire knowledge and skills for lifelong participation in physical activity that contributes to optimal health benefits (Metzler et al., 2013)8. A CSPAP intends to provide expanded opportunities for physical activity beyond regularly scheduled Physical Education time—including before, during and after school, as well as opportunities outside of school (e.g., at home and in the community.
Health Optimizing Physical Education (HOPE) aims at developing the public health goal for students to accrue adequate amounts of current physical activity and be prepared for an active lifestyle in adulthood. It involves all the socio- ecological factors that affect the child so as to achieve the optimal health benefits. They are Individual, Interpersonal, Organizational, Community and Public Policy (Metzler et al., 2013)8.
Another teaching curriculum called Teaching Games for understanding (TGFU) are developed in America to elicit the players`/students’ tactical awareness and skill development from situated learning experience enabled by the teacher/coach/Physical Educationist. Naomi Hart (2010)9 discussed the model of TGFU as (Figure 3): a) Game Participation: The learning in Physical education starts with games participation in a modified game or real games. At this stage the formative assessment takes place, b) Game Appreciation: Through game play students are given the chance to enjoy and experience of learning, c) Tactical Awareness: Through game play students are encourage to identify attacking and defending strategies and implements successful tactical action, d) Making appropriate decision: In dynamic situation students make real time decisions. These could be with or without the ball, e) Skill execution: Do the students have the ability to convert their decisions into actions? Can they execute the correct skill to enable their team to succeed? F) Performance: Students take part in full or modified game applying all new knowledge and skills. This is the time for summative assessments. A detail on TGFU was described by Steven Stolz, Shane Pill (2014)10 and Linda Griffin and co-author (2005)11in the Physical Education Review journals.
Discussion
According to U.S. Olympic Committee, the university’s athletes are dominating the U.S. Olympic Team in which the U.S. has competed since 1912. This Olympians are not born, but they are made through their lifetime participation in research based PE program from school. In the west, children are taught the importance of physical education from the grass root level. A progressive structure is being followed so as to inculcate the basic techniques of movements, skills and a sense of fitness right from early childhood.
PE program has its unique benefits and its program fits into each other. An example of how a progressive structure PE curriculum intertwine and integrate one into other from early childhood, kindergarten and grade 3-6 program can be explained by teaching the skills of throwing and catching. In early childhood, a child would be exposed to the throwing and catching of a balloon (Figure 4a). The weight of descent is slower and gives time to the child to reflex while trying to catch the balloon. As the child gets more comfortable with the balloon, the object is replaced with a scarf (Figure 4b). Since the scarf falls quicker to gravity, the personal space of the child is maintained and safely guarded. In the program for the age group ranging from kindergarten to Grade 2, a slightly heavier object (for example a fluff ball) is used. Since the fluff ball (Figure 4c) is heavier, it will fall faster, which is again age specific and is safer for the child. Therefore, instead of using a big ball to teach children of this age group, a fluff ball is safer and will save their finger from getting hurt. The primary focus is to inculcate gross motor skills in this age group. This activity is a progression of the movement techniques learnt by the children in their early childhood. In this level, the movement is more precise and prepares them to learn higher techniques as the basic techniques of movements will help them in formulating to a better motor program.
In the curriculum for the age group of grade 3 to 6, the action is more defined into a more appropriate throwing action. At this stage, tennis ball (Figure 4d), is used. As they get more comfortable with the tennis ball, the children can be introduced to partners (Figure 4e) so that the skills of the children are groomed and developed to improve their potential, similar to that of playing cricket wherein the ball is actually thrown back and forth to learn catching and throwing skills. By the time the child goes to the entire progression through the PE program and the child skills is hold at the highest level to be implemented in the future.
Conclusions
New curriculum, age appropriate equipments, teaching techniques, etc should be introduce in all the schools to make the child engage in physical activity for at least 60 minutes per day. However, the use of innovative equipments, strategies and curriculum in PE classes must be research-based and need for the children. Physical education program should be one of the major subjects in schools curriculum since the physical educator has a great role to play in the grass root level to groom a child into making a potential and highly skilled professional.
Acknowledgements
Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. Special thanks to LEAPSTART, Bangalore, India for sharing us innovating ideas for physical education program.
Source of Funding
None
Conflict of interest
None
Englishhttp://ijcrr.com/abstract.php?article_id=2285http://ijcrr.com/article_html.php?did=2285[1]) Sallis, J. F., and McKenzie, T. L. (1991). Physical education’s role in public health. Research Quarterly for Exercise and Sport, 62,124–137.
[2]) Stewart G. Trost, Hans van der Mars (2009), Why We Should Not Cut P.E. Health and Learning Pages 60-65 Volume 67 | Number 4
[3]) SPARK. How SPARK Aligns with NASPE’s National Standards for Physical Education. mailto:http://www.sparkpe.org/standardsNASPE.pdf. Accessed on 15th February, 2017
[4])Indiatoday (PTI). Grukul trust launches school sports promotion foundation. http://indiatoday.intoday.in/story/grukul-trust-launches-school-sports-promotion-foundation/1/469116.html. Accessed on 15th February, 2017
[5])http://www.sparkpe.org/physical-education-resources/academics-physical-activity/#sthash.iz3305Xg.dpuf. Accessed on 21th February, 2017
[6] ) Bevans, K. B., Fitzpatrick, L.-A., Sanchez, B. M., Riley, A. W., and Forrest, C. (2010). Physical Education Resources, Class Management, and Student Physical Activity Levels: A Structure-Process-Outcome Approach to Evaluating Physical Education Effectiveness. The Journal of School Health, 80(12), 573–580. http://doi.org/10.1111/j.1746-1561.2010.00544.x
[7] ) Judith E. Rink, Tina Hall, Lori Williams (2010). Schoolwide physical activity: A comprehensive guide to designing and conduction programs. Human Kinectics News and Excerpts. http://www.humankinetics.com/excerpts/excerpts/the-role-and-responsibilities-of-the-physical-education-teacher-in-the-school-physical-activity-program
[8] ) Mike Metzler, Thomas McKenzie, Rebecca Ellis, et al., (2013).Health Optimizing Physical Education (HOPE): A New Curriculum Model. Journal of Physical Education, Recreation and Dance, Volume 84, Issue 4, 2013 pages 41-47, DOI:10.1080/07303084.2013.773826
[9]Naomi Hart (2010), Teaching Games for Understanding. www.naomihartl.weebly.com/blog/archives. Accessed on 20th April, 2017
[10] ) Steven Stolz, Shane Pill (2014)Teaching games and sport for understanding. European Physical Education Review; Vol 20, Issue 1, pp. 36 - 71. doi: 10.1177/1356336X13496001
[11]) Linda Griffin, Joy Butler (2005) Teaching Games for Understanding. Saskatchewan Physical Education Association.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30General SciencesCharacterization of Gem and Abrasive Variety Corundum from Wyra Area of Khammam Schist Belt, Telangana, India, Using Advanced Spectroscopic Techniques
English1216S. U. Shyam KumarEnglish N. Shadakshara SwamyEnglishCorundum of varying qualities occurring in association with Khammam schist belt are studied with respect to their Geochemical and Mineralogical aspects. Studies have been carried out using the advanced spectroscopic techniques namely, Photoluminescence, UV-Visible, FT-IR, Raman and EDXRF analysis. The results obtained from the studies shows the predominance of colour causing element Cr and other associated elements like, Ti, V and Ga in Red variety of corundum than the abrasive corundum crystals, while Fe is found predominant in colourless or abrasive variety. From EDXRF studies the presence of impurity ions in the crystal lattice is quantified and its effect on the corundum crystal has been ascertained.
EnglishCorundum, Ruby, Telangana, Spectroscopy, Cause of colourINTRODUCTION AND GEOLOGY
In and around Wyra taluk in the Khammam district of Telangana, India, occurrences of corundum of abrasive variety and rarely of gem variety are observed. The study area lies in the Khammam schist belt (KSB) which is a continuation and part of the Nellore-Khammam Schist belt (NKSB) (Ramam and Murthy, 1997).
The main lithology of the area include amphibolite, schists, and gneisses of varying compositions, anorthosites, pegmatites and quartz veins, corundum bearing magnesium and aluminum rich pelitic granulites and hornblende pyroxene granulites (Ramam and Murthy, 1997; Patil, 1973). Metamorphosed pelitic, psammitic and calcareous sediments are represented by garnetiferous biotite- muscovite-kyanite-chlorite schists, kyanite and sillimanite bearing meta - pelitic schists, calc -silicate rocks, quartzites, fuchsite, and magnetite quartzite. The meta-sedimentary unit is tectonically emplaced below a metavolcanic unit made up of quartz-chlorite schists, hornblende schists and tonalitic gneisses (Ramam and Murthy,1997; Leelanandam and Narasimha,1988; Narayan and Pavanaguru, 2013).
The main host rocks for corundum in the study area are sillimanite-corundum schist and kyanite schists. These pelitic schists hosting corundum are found to be in contact with altered basic bodies or pegmatitic vein. Occurrences associated with pyroxene granulites and gneisses are also seen in some localities of the Wyra taluk. Similar Geological setups hosting corundum have been reported from the study area (Patil, 1973; Rao,1981; Narayan and Pavanaguru,2013). However, corundum crystals for the present study are obtained from placer concentrations seen to occur locally in the soil profile.
Samples of gem variety as well as abrasive variety of corundum are collected from these areas and, 2 samples of contrasting qualities are selected for studies.
MATERIALS AND METHODS
Samples collected from the study area are analyzed based on its visual appearance and it has been grouped on that basis. Two Corundum samples are sorted from them based on the colour and gemological quality so that, one sample is of abrasive quality with colourless nature ( C1, Fig.1) where as the other one is of semi precious variety with pinkish red colour (C2, Fig.2).
Both the samples C1 and C2 after preliminary cleaning and acid treatment were cut into suitable size and then subjected to the following analyses namely, UV-Visible Spectroscopy,
Photoluminescence spectroscopy, FT-IR Spectroscopy, Raman Spectroscopy and EDXRF studies.
RESULTS AND DISCUSSIONS
The spectra obtained for both the gem and abrasive variety samples using different advanced spectroscopic analyses are displayed with contrasting colours. The UV-Visible spectra
obtained for the samples in the wavelength range of 200 nm to 600 nm and Photoluminescence peaks in the wavelength range of 600 nm to 800 nm are shown in Figure.3 and Figure.4 respectively. Vibrational Spectroscopic studies on the samples using the Fourier Transform Infrared (FTIR) Spectroscopy and the Raman Spectroscopy are discussed
separately. Figure 5 and Figure 6 depicts the spectra obtained using FTIR Spectroscopy in a wavelength range between 4000 cm-1 to 500 cm-1 and Raman Spectroscopy in Raman
shifts ranging from 200 cm-1 to 800 cm-1, respectively. Results obtained from EDXRF Studies are discussed and correlated with the observations made from other Spectroscopic
techniques used in the research.
UV-Visible and Photoluminescence
Spectroscopy
The UV-Visible spectra of the two samples are observed in the wavelength range of 200 to 600 nm (Figure.3). The spectra show significant difference in the peaks obtained at certain wavelengths. The peaks shown by the C2 sample are of more intensity and magnitude compared to that of C1 sample, indicating the presence of certain additional elements in C2, which has favored in imparting pinkish red colour. Peaks in the range of 200 to 250 nm are more densely spaced in the C1 variety of corundum sample. A prominent peak at 241 nm is seen in C1. The hydroxylated ions present in samples correspond to absorptions in this wavelength. Such ions are hence present in higher concentration in the C1 variety.
Correlation of the absorbance peaks clearly shows a contrast in the peaks at wavelengths corresponding to Fe and Cr, the usual colour causing ions in ruby variety of corundum. Some of the studies (Nassau et al.,1983; Jatin and Pratima,2015) has shown that, the wavelengths that are usually absorbed by Fe ions falls under the range of 380 to 400 nm whereas, peaks are observed to be present at 401, 461, 467, 475 and 535 nm when the Cr ions shows its prominence .
In the absorbance spectra of the samples, C1 is having peaks of very minute magnitude in the wavelength range corresponding to Fe ions and it doesn't shows any absorbance in the range of Cr ions. It is identified that, the peaks in this range, corresponding to Fe and Cr are present in C2 with a comparatively appreciable magnitude, which justifies the cause of colour in the C2 sample compared to C1.
The Luminescence peaks are observed for both of the samples in an excited state at 401 nm (Figure.4). Luminescence peaks lies in the wavelength range between 600 to 800 nm. The dual peak at 692 and 694 represents the presence of Cr chromophores in both the samples (Nassau et al.,1983; Beran and Rossman et al.,2006; Jatin and Pratima,2015).
Even though C1 sample has a significant peak at the above mentioned wavelengths, the magnitude is much lesser than that of C2. The very high magnitude of the peaks obtained for C2 samples substantiates its cause of colour due to prominent presence of Cr.
FT-IR Spectroscopy
Fourier Transform Infrared Spectroscopy or FT-IR Spectroscopy data of the samples is observed (Figure.5) in a wavelength range between 4000 cm-1 to 500 cm-1. The spectra seen between 500 to 1000 cm-1 is of the same fashion with peaks at common wavelengths. Both the samples show peaks corresponding to same wavelengths, even though the C1 sample has a comparatively lesser magnitude. Therefore, these peaks are generally identified to be caused by the Alumina present in the crystal.
There are no considerable peaks seen in the C1 sample after 1000 cm−1. Twin peaks observed in C2 sample at 1991 and 2100 cm−1. Similar peaks are observed (Cowley,1969; Kloprogge et al.,2004; Jatin and Pratima,2015), indicating the presence of hydroxylated form of Alumina, corresponding to high temperature environment and the peaks seen in the wavelength range towards higher energy part is all corresponding to different hydroxylated ions in the crystal lattice. These are caused by the vibration modes related with the bending and stretching of –OH group bonds in the crystal lattice and the mineral groups that may be present cannot be always presumed to be due to the –OH association with Alumina groups. Such hydroxylated groups are similarly observed (Beran and Rossman,2006; Kloprogge et al., 2004) to be due to the presence of other elements like Ti, Fe or V present in the crystal lattice in traces .
Raman Spectroscopy
Raman spectroscopy is depended on the vibrational modes of the substances subjected to analysis. Corundum is reported to have 18 vibrational modes where 7 are Raman active, 6 are Infrared active while the rest are neither Raman nor Infrared active and unique vibrations for Corundum are usually observed at 378, 418, 432, 451, 578, 645 and 751 cm−1 (Porto and Krishnan,1967;XU J-A et al., 1995; Jasinevicius, 2009).
Individual spectra of both the samples C1 and C2 are well observed (Figure.6). The spectra obtained for C1 sample is devoid of any considerable peaks and is almost following a plain linear fashion.
In contrary, the spectra of C2 sample is showing a notable peak at 417 cm-1,a minute peak at 750 cm−1 and a very minute one at 577 cm-1.
These peaks correspond to the typical fundamental vibrations evidenced in corundum, which are also believed to be associated with Alumina.
Geochemical Analysis
Characterization of the two variety of corundum in terms of chemical composition is highly essential in determining the cause of colour and other allied properties in Gemological perspective. The average weight percentage in terms of oxides of Al, Ca, Ti, V, Cr, Fe, Ga and Zr are obtained from the Energy Dispersive X Ray Fluorescence (EDXRF) Spectroscopy.
The presence of other oxides in the C1 samples is estimated to be within 0.8 weight percentage and the crystal is mere corundum with higher content of Al203 alone, even though 0.3 weight percentage of Fe2O3 is also observed. The C2 sample shows 95.3 weight percentage of Al203 while other oxides constitutes for an average weight percentage of 3.179 out of which,Cr2O3 predominates with 1.882 by weight percentage followed by, TiO2, CaO and ZrO2.
CONCLUSIONS
Present study indicates subtle differences in the two samples. The UV-Visible spectra shows continuous peaks in the wavelength range from 401 to 535 in the C2 sample, while it remains absent in that of C1, revealing the predominance of Cr ions in C2 sample. The emission spectra due to Photoluminescence, has prominent dual peaks in the wavelengths 692 and 694nm in C2 sample while these peaks are present in a very less magnitude in the C1 sample. This accounts to the prominence of Cr chromospheres in C2 sample. Peaks obtained from the FTIR spectra, in the higher energy level prior to 1000cm-1, throws light on the existence of several hydroxylated ions in the crystal, along with Alumina in the C2 sample while, peaks in this range remains absent in C1. The Raman spectra shows peaks at the typical fundamental vibrational modes corresponding to their respective Raman Shifts cm−1 at 417, 750 and 577, in the C2 sample, while C1 sample is showing a spectra devoid of such peaks. EDXRF data substantiated the results obtained from other advanced spectroscopic analyses as it confirms that the presence of Cr2O3 and TiO2, which are desirable constituents of quality rubies is predominant in C2 sample, while, the quantity of Fe2O3 is higher in C1.
This study has been proven helpful in characterizing and determining the parameters and features present in the two varieties of samples collected from the study area, that are responsible for causing colour in them.
ACKNOWLEDGEMENT
The authors are thankful to the Chairperson, Department of Geology, Bangalore University, Bangalore, for providing research facilities. We are grateful to Dr. M.D. Sastry, Head, R and D and Shri.K.T. Ramachandran, Chief Gemologist and Executive Secretary, of the Gemological Institute of India, Mumbai, for extending facilities for carrying the advanced spectroscopic analyses at their R and D Laboratory.
Englishhttp://ijcrr.com/abstract.php?article_id=2286http://ijcrr.com/article_html.php?did=2286
BERAN, A. and ROSSMAN, G.R. (2006) OH in naturally occurring corundum. European Jour Mineral, V.18, PP.441-447.
BHAGAVANTAM, S and VENKATARAYUDU, T. (1939) Raman effect in relation to Crystal Structure. Proc. Ind. Acad. Sci, V.9, PP.224-58.
COWLEY E.R.(1969) Symmetry properties of normal modes of Vibration of Calcite and Corundum. Can J. Phys, V.47, PP.1381-91.
JASINEVICIUS, R. (2009), Characterization of vibrational and electronic features in the Raman spectra of gem minerals. MSc Thesis, Grad. College, University of Arizona, 147 p.
JATIN KUMAR SINHA and PRATIMA KUMARI MISHRA (2015) Spectroscopic and Microstructural Studies of Ruby Gemstones of Sinapalli, Odisha. Geol. Soc. India. Indian. Jour. Vol. 86. PP.657-662
KLOPROGGE, J.T., HICKEY, L. and FROST, R.L. (2004) FT-Raman and FT-IR spectroscopic study of synthetic Mg/Zn/Alhydrotalcites. Jour. Raman Spectros., V.35, PP.967-974.
LEELANANDAM, C. and NARSIMHA REDDY, M (1988)Precambrian anorthosites from Peninsular India- Problems and Perspectives. Indian. Jour. Geol, V.60, PP.111-136.
NARAYAN SANGAM. and PAVANAGURU, R (2013) Geology of Corundum Occurrances in Parts of Khammam Schist Belt. IJSRP. V.3, 3P.
NASSAU, K. (1983) The Physics and Chemistry of Color: the 15 Mechanisms. Wiley, New York, PP 451-462.
PATIL R. D. (1973) Report On The Investigation For Corundum Near Gobbagurti, Khammam District, Andhra Pradesh. Geol. Surv. India, unpublished report. PP 1-4
PORTO, S.P.S. and KRISHNAN, R.S (1967) Raman Effect of Corundum. Jour. Chemical Physics, V.47, PP.1009-1012.
RAMAM, P. K. and MURTHY, V. N, (1997) “Geology of Andhra Pradesh”. Geol. Soc. India. Jour. Vol. 78,245P.
RAO, N. A. (1981) “Report On The Investigation For Corundum In Khammam District, Andhra Pradesh”. Geol. Surv. India, unpublished report. PP.1-4.
RICHET, P., GILLET, P., PIERRE, A., BOUHIFD, M.A., DANIEL, I., FIQUET, G. (1993) Raman spectroscopy, x-ray diffraction, and phase relationship determinations with a versatile heating cell for measurements up to 3600 K (or 2700 K in air). Jour. Appld. Phys, V.74, PP.5451-5456.
XU J-A, HUANG E, LIN J-F, XU LY (1995) Raman study at high pressure and the thermodynamic properties of corundum: Application of Kieffer’s model. Ameican Mineralogist, V.80, PP.1157-1165.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30General SciencesGeo-Environmental Etiology of Allergic Disorders and its Impact on Human Health in Sopore J and K, State, India
English1722Manzoor A. WaniEnglish Sheraz A. LoneEnglish Bashir A. LoneEnglish Ishtiaq A. MayerEnglish Nisar A. WaniEnglishThe present study aims to comprehend the causes responsible for the occurrence of allergic disorders and access their impact on human health in peculiar climatic conditions of Sopore tehsil district Baramulla, J and K, State, India.
Objective: To understand the dissemination phenomenon of known allergenic pollen types causing allergic disorders among the people of the selected sample area in Jammu and Kashmir State, India.
Methodology: An effort is being carried out to study airborne pollen and assess their impact on human health in a peculiar climatic condition of Sopore tehsil of district Baramulla in the north region of Kashmir valley of Jammu and Kashmir state by using both primary as well as secondary source of data in order to envisage the relation of environmental factors on the allergic disorders with their impact on the human health.
Results: In the present study 1413 cases of allergic disorders were recorded in the district hospital, sub-district hospital and primary health centers from the year 2010-2015, out of which 57.18 percent were constitutes males. Cases like rhinitis (35.52 percent), conjunctivitis (28.09 percent), dermatitis (15.28 percent) and bronchial asthma (21.08 percent) were recorded in the study area.
EnglishTemperature variability, wind velocity, pollens, Allergens, Pulmonary disordersIntroduction
The prevalence rate of respiratory and allergic diseases such as asthma and rhinitis have increased dramatically to epidemic proportions worldwide. The World Allergy Organization (WAO) raises concern of this health hazard and highlights the role of climate-related health impacts on mortality and acute morbidity of respiratory disorders due to extreme meteorological events. Pollens are by and large considered as causative agents of respiratory allergy disorders in any geographic area1. It has been established that the frequency and incidence of allergic pollen is high in areas of low relative humidity, modest temperatures and maximum rainfall2. Pollen dispersal is facilitated by dry weather with low relative humidity as the pollen particles being light and dry disperse in the air with less constraint. Wind velocity and wind direction plays an important role in pollen release and in its transportation during flowering phase5. It has also been observed in Kashmir (India) that besides high temperature and low relative humidity the pollen dispersion from the exotic Australian populous tree enhances the liberation and distribution of pollen resembling light cotton flakes in the atmosphere and are capable to trap the allergen elements (pollen) which unwillingly enters into the human organs through inhaling organs thus irritates the eyes. Air borne allergen pollen flora are widely distributed all over the India with no exception to Jammu and Kashmir which enjoys frequent weather changes thus plays a major role in the dispersal of allergen pollens inhaled by human when exposed to open environment7.
Study Area
The study has been carried out in Sopore tehsil,a second order administrative unit of Baramulla district of Jammu and Kashmir State (India). Sopore known as Suyyapur in antiquity is a prosperous geographic region in the state Jammu and Kashmir. It is located at 34030'N latitude, 74047'E longitude, constitutes an area of 320sq.kilometers and is known as “Apple town of Asia”; because of its horticultural produce. The area offers opportunity to the larger people of district to earn livelihood in horticulture/agricultural related activities. Therefore it represents the geographic region of north Kashmir in terms of population composition and its characteristics.
Data base and Methodology
The data was collected from district hospital, sub- district hospital and primary health centers located in the study area. The climatic data was collected from meteorological department Srinagar. Mean monthly temperature, average annual rainfall and average wind velocity from year 2010-2015 were taken into consideration to check seasonal trend. The other factors interlinked with allergic disorders were also taken into consideration such as flora of the region, occupation structure etc. After disseminating the meteorological data with medical records the impact of allergic disorders on the human health with seasonality were seen on both genders as well as on different age groups in the study area.
Results: Different parameters such as meteorological factors (Temperature, rainfall, wind velocity) from year 2010-2015, occupational structure, and flora were interlinked with various allergen antigens and their impact was seen on the human health in the study area. The results are shown in separate tables listed.
Discussion: In this study it was found that allergen pollen along with various meteorological parameters (Temperature, Rainfall and Wind velocity), flora and flowering period time, occupational structure have a strong bearing on the human health. In the study area about 1413 cases of allergic disorders were recorded in the district hospital, sub-district hospital and primary health centers from 2010-2015, out of which 57.42 percent of the cases of allergic disorders were related to men of all age group. The higher percentage of males was mainly attributed to nature of work and exposure to open environment in agricultural fields. About 46 percent such cases with rhinitis, conjunctivitis, dermatitis and bronchial asthma were recorded in the month of March- June during. 2010-2015(half decade). 66.38 percent the case were in the age group of 21-50 years being the more potent labour class and is peak season of as agricultural activities in Kashmir valley. The previous nearly six months remains dormant for such activities due harsh winter and inclement weather conditions in the valley. Due to the onset of spring from 21st March in the state people start their work in the fields (Agricultural, Horticulture etc.) in order to generate their daily livelihoods. All the factors favors the allergic disorders such as temperature for the blooming of allergenic pollen flower, wind and moisture require transporting these allergen pollen spores from lower layer of atmosphere to hitting targets mainly (human population).
Conclusion: The study highlights the impact of allergic pollens along with different meteorological factors on human health. Clear evidences are traced from the study area which shows a relationship of allergen pollens and human health. Diseases such as Rhinitis, Conjunctivitis, Dermatitis, and Bronchial asthma were recorded in both field as well as from medical records witnessed during different seasons in the study area of all ages irrespective of gender. The study revealed that local factors especially presence of allergen flora, nature of work mainly contributed to etiology of allergic disorders.
Suggestions:
Avoidance is the best treatment, but it is very difficult to follow as it is not possible to avoid pollen completely as majority of the life dependence is with agriculture economic activities. General masses can be advised to use air masks thereby reducing the inhaling of allergens. Another method is immunotherapy where extract of a particular antigen or mixed antigen to which patients are allergic is given in very low concentrations initially and then the concentration is increased slowly. As a result, a patient develops antibodies against the specific antigens. More importantly government must to take necessary measurements before introducing any new exotic flora species so to be eco-friendly in nature.
Acknowledgement:
I wish to express my cordial gratitude to my esteemed supervisors Prof. Ishtiaq A. Mayer, for his meritorious, sympathetic and encouraging guidance. Words fail me to explain his inspiring guidance, immense interest, supportive and affectionate behavior. I would like to express my special thanks to my co-authors without their role and self dedication it would have not been possible, every effort from them is priceless. My special word of thanks goes to Dr. Nisar Ahmad, Dr. Marukh Hameed (Assistant professor in Human Genetics in SKIMS). I am grateful to all the scholars, authors, publishers for all those articles, journals and books where the literature for this study was taken and discussed, I wish them all the best. Last but not least words fail me to translate appropriately my gratitude and deep sense of inner feelings about the inspiration and help provided by my loving family.
Table 1: Occupational structure
Tehsils
Cultivators
Agricultural laborers
Household industry workers
Others
Sopore
23,682
5,010
2,901
30,730
Source: Census department India
Table 1 shows the occupational structure, the main occupation of district Baramulla in general and Sopore tehsil in particular comprises of agricultural cultivators. Both male and females spend most of their time in their field. The peak season of agriculture starts from month of March and it continue till October-November. From month of March the flowering of trees emerge which continues till the month of June. Therefore during pollination various pollens dissimilate with the help of wind.
Table 2: Mean monthly meteorological data of Kashmir valley (average 2010-2015)
Months
Temperature 0celcius
Maximum Minimum
Average. Rainfall
(millimeters)
Wind velocity
Km/hour
January
7.3
-3.2
98.1
2.2
February
8.8
-0.2
188.9
2.1
March
17.0
3.4
114.1
3.4
April
20.0
7.8
171.6
3.4
May
23.6
10.1
114.8
3.0
June
27.9
13.8
47.6
2.5
July
30.9
17.0
64.5
2.3
August
30.3
17.5
77.8
2.3
September
27.1
13.2
102.9
2.5
October
22.7
5.6
59.9
2.1
November
18.0
1.3
22.3
2.1
December
10.3
-1.8
66.7
2.2
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Chanda, S and Sarkar, P.K. 1972. Pollen grains as a causative agent of respiratory allergy with reference to aeropalynology of Greater Calcutta. Teans. Bose. Res. Inst, 35:61-67.
Davis, R.R and Smith, L.P.1997. Weather and the grass pollen content of the air. Clinical allergy, 3:95-108.
Eng, K and Knox, R.N. 1994. Seasonal distribution of pollen in the atmosphere of Melbourne: an airborne pollen calendar. Int. J. Aerobiol, 2:51-5.
Gennaero, D and Stephan, T.2015. Meteorological conditions, climate change, new emerging factors, and asthma related allergic disorders. A statement of the world allergy organization. World allergy organ, j. 1:25.
Gupta, K.D and Singh. 1965. Pollination calendars of allergenic plants of Bikaner (Rajasthan). Botnical survey of area. J.Ass. Phys. India, 13:143-175.
Mervi, H.1992. Long-distance transport of Betula pollen grains and allergic symptoms. Aerobiologia. 2: 231-236.
Munshi, A.H.2000. Flowering calendar of grasses in Srinagar, Kashmir Himalaya (India). Aerobiological, 16:449-452.
Nair, P.K and Joshi, A.P.1986. Airborne pollen, spores and other plant material of India- A survey Publ. CSIR Centre for Bio-chemicals, Delhi and National Botanical research Institute, Lucknow 200-224.
Nisar, A. Wani.2009. Specific Immunoglobulin E (IgE) and skin prick test (SPT) Responses against battery of pollen antigens in allergenic subjects of Kashmir-India, African journal of Basic and Applied Sciences, 2 (1-2):30-32.
Ron, M. E and M. Perzanowski, Plats. 2003. Different sensitization profile for asthma, rhinitis and eczema among 7-8 year old children: Report from obstructive lung diseases in northern Sweden studies. Pediatric Allergy Immunol. 14:91:99.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30HealthcareAssessment of Genome Integrity in Mitochondria of Testis Tissue, Cell Proliferation and Topoisomerase Assay in invitro Condition
English2326Eswari BeeramEnglishAim: Assessment of genome integrity in testis and to explain the reasons for infertility with the drug metosartan.
Methodology: Comet assay with freshly isolated mitochondria, cryopreserved mitochondria and cryopreserved tissue mitochondria and cell proliferation assay by dye exclusion method and topoisomerase assay in fresh and deep freezed DNA.
Results and Discussion: Comet assay of isolated mitochondria both from fresh and cryopreserved samples was examined by fluorescence microscope, which revealed that cryopreservation of testes mitochondria samples lead to injury to the tissue and DNA fragmentation. Topo isomerase assay has showed that increased incubation leads to DNA fragmentation and cell proliferation assay with the drug has showed the 0% viability of the cells by trypan blue. So, in vivo studies are necessary to confirm whether the drug is safe or not.
Conclusion: Assessment of genome integrity lead to the finding that drug was not safe it resulted in so many difficulties in invitro condition.
EnglishMitochindria, Genome, Cryopreserved, TopoisomeraseIntroduction:
Comet assay is a combination of both agarose gel electrophoresis along with fluorescent microscope for detecting the DNA strand breaks in the nucleated cell. The extent of DNA migration is measure of damage in the genome of the cell. This is a simple technique as it doesn't require any radioactive labelling compounds and also problems associated with disposal. It can be more useful in case study of DNA damaging agents. The damaged DNA migrates as comet. The comet head consists of high molecular weight DNA and tail consists of the DNA single strand fragments. Applications of comet assay includes diverse things(1), in which change of lysis buffers and DNA stains resulted in different achievements like use of neutral buffer to find the double strand breaks whereas use of buffer containing specific endonucleases can used to find damage at base level. Here in my study I have taken neutral buffer to detect double stranded breaks in mitochondria isolated in pure form cryopreserved before the comet assay was performed. As the mitochondria is present in midpeice of sperm and responsible for motility of sperm , So studies on mitochondria genome is necessary as decreased motility of sperm leading to problems in fertility.
Cell proliferation assay is important to know the abnormal proliferation due to certain agents. Simple techniques like MTT assay and trypan blue assay is used to know whether the cells are normal or not (6). AS MTT assay of control gives the same result after the dye has entered in to the dead cell and So, useful to perform the trypan blue as the dye was taken by the non viable cells itself. In this study I have used metosartan to know the cell viability in vitro using trypan blue. The cells were dead and matched with the results of aniline blue staining.
Mammals express two types of topoisomerases alpha and beta which are homologous but differ in the sub cellular location of the enzyme. Similar to topoisomerase I and III, topoisomerase II also involved in relieving the topological constraints of DNA(2). Topoisomerase is homodimer binding to the clamps of DNA polymerase and the strand was cleaved by topoisomerase first using the ATP, the cleaved strand was passed through the gap, re ligated and the strands are released by hydrolysis of ATP after the release of ADP molecules.
In general sense DNA is unwinded in the replication in order to pass the genetic information or for transcription of the mRNA strands. In both the steps DNA winding is necessary but focussing on DNA replication there will be formation of positive supercoils and makes the the fork movement difficult an under winding of the DNA behind the fork also leads to torsional stress in the replication and leading to formation of catenated DNA . As we know helicases separates the two strands but not involved in the removal of torsional stress but here, the topoisomerases came in to role as that mentioned in the previous paragraph.
Topoisomerase II is also responsible for maintenance of chromatin integrity as supercoiled that means folded supra molecular structures both in fresh and frozen samples. The present aim is to know the genome integrity in the treated sample as in case of proliferation assay and fresh and frozen samples by topoisomerase assay and in fresh and cryopreserved samples in comet assay.
Materials and methods:
Cell proliferation assay using trypan blue :( 6)
1. The cell suspension was prepared in a balanced salt solution (e.g., Hanks' Balanced Salts [HBSS], and 0.5 ml of 0.4% Trypan Blue solution (w/v) was transferred to a test tube. To it 0.3 ml of HBSS is added along with 0.2 ml of the cell suspension (dilution factor = 5) and mixed thoroughly and Allowed to stand for 5 to 15 minutes after addition of metosartan drug to the suspension.
2. After proper incubation by using Pasteur pipette or any suitable device transfer the trypan blue- cell mixture suspension to both the chambers by just touching the tip of the pipette to the edge of the chamber and allowing it to fill by capillary action and overfilling or under filling was avoided.
3. Starting with the chamber 1 count all the four corners squares and repeat the procedure for chamber 2 also. Nonviable stains blue.
Cryopreservation of tissue:
Liquid nitrogen was brought from S.V veterinary university and the tissue is homogenated in water and exposed to liquid nitrogen vapours and immersed in liquid nitrogen for 3 days and mitochondria was isolated after the cryopreservation and comet assay was performed... The isolated mitochondria was also cryopreserved, and processed in the same manner
Isolation of mitochondria from testes:(7)
After the sacrificing the animal, testes were removed, decapsulated and minced in isolation medium 250mM sucrose, 0.2 mM EGTA, 0.1mM EDTA,5mM HEPES-KOH (PH 7.4) and 0.1%defatted BSA. The tissue was homogenated in isolation medium and the homogenate was centrifuged at 10,000Xg for 10min and the supernatant was recentrifuged at 10,000Xg for 10min. The pellet was resuspended and repelleted twice in the isolation medium omitting EGTA, EDTA, and defatted BSA.
Comet assay: (3)
Standard comet assay was performed..100l of the isolated mitochondria was mixed with 1000l of 0.8% agarose and 1000l of suspension was spread on the slide and kept in ice for 45min. After casting the slides was immersed in lysis buffer(2.5% SDS in 45 mM Tris-borate, 1 mM EDTA, pH 8.4) for 20min and kept in electrophoresis buffer for 5min. The electrophoresis was carried with TBE buffer at 2v/cm. Ethidium bromide staining was performed and visualised using fluorescent microscope.
Isolation of DNA from testis tissue :( 4)
Cells were pelleted and the suspension was collected. To the supernatant lysis buffer (50mM tris-cl, PH8.0, 100MEDTA,0.125%SDS) was added and the solution was incubated in boiling water bath at 550c for overnight. To this 1ml of mixture of phenol :chloroform : isoamyl alcohol was added and centrifuged at 10,000rpm for 15min. Aqueous phase was collected and washed with 3ml of phenol: chloroform : isoamyl alcohol. Aqueous phase was collected and chilled ethanol was added and upper aqueous phase was separated and centrifuged at 10,000rpmfor 10min and the pellet was resuspended in 100l of water.
Topoisomerase assay :( 5)
The above isolated DNA was suspended in topoisomerase reaction buffer 10X ( 200mM tris-cl, PH:7.5, 100mM Mgcl2, 10mMATP, 10mM EDTA, 10mM, β-mercaptoethanol, 1.5 M KCL,300g/ml BSA. The suspension was incubated for 1-3hrs and agraose gel electrophoresis was performed with TAE buffer.
Results:
Comet assay of fresh and cryopreserved testis sample mitochondria has given the conclusion that cryopreservation may result in distortion of shape, micronuclei formation which may result low motility of spermatozoa therefore leading to infertility. Incase of cryopreserved mitochondria after isolation showed less distortion and absence of micronuclei and also comets formed was similar to fresh sample.
In fresh sample the comets was up to acceptable extent than compared to the cryopreserved mitochondria testes samples.
In the DNA isolated from the fresh tissue DNA fragments was not found where as in refrigerated samples treated with topoisomerase showed supercoiled structures and also the fragmented molecules which migrated to the longer distance in the gel. With increase in time period the intensity is also increased representing the cleavage.
Calculations:
Cell Counts - Each square of the haemocytometer, with cover-slip in place, represents a total volume of
0.1 mm3 or 10-4 cm3. Since 1 cm3 is equivalent to approximately 1 ml, the subsequent cell concentration per ml (and the total number of cells) will be determined using the following calculations:
Cells Per mL = the average count per squarex dilution factorx 104 (count 10 squares) If the average count per square is 15 cellsx 4x 104 = 6x 105 cells/ml.
Total Cells = cells per mlx the original volume of fluid from which cell sample was removed.
6x 105 (cells/ml)x 10 ml (original volume) = 6x 106 total cells.
Cell Viability (%) = total viable cells (unstained)total cells (stained and unstained) x 100. If the average count per square of unstained (viable) cells is 0, the total viable cells = [0x 4x 104] viable cells/mlx 10 ml (original volume) = 0viable cells. Cell viability (%) = 0 (viable cells) 6x 106 (total cells)x 100 = 0% viability.
From the cell proliferation assay it was clear that the drug incubation for 30min lead to 0% viability
Discussion:
Cell proliferation assay, topoisomerase assay and comet assay was used in the study for assessing the genome integrity. Comet assay was performed by so many people but for mitochondria is a new thing. Mitochondria isolated from the cryopreserved tissue showed micronuclei formation which indicates that DNA was fragmented and from fresh tissue revealed less by comets formation. Cell proliferation assay revealed that treatment with drug for 30min resulted in 0% viability. So, the drug was not safe and it should be confirmed with in vivo experiments also, in order to come to a conclusion.
Conclusion:
The drug metosartan resulted in formation of micronuclei in mitochondrial genome and also decreased viability by trypan blue, topoisomerase assay has revealed that with incubation the DNA was fragmented resulting in the bands as supercoiled and fragmented DNA.
Acknowledgement:
Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Conflict of interest:
No conflict of interest
Source of funding:
UGC- CSIR JRF
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1. Peggy L Olive and Judit P Banáth .The comet assay: a method to measure DNA damage in individual cells VOL.1 NO.1 | 2006 | 23-29.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30HealthcareModulation of Antioxidant Status of Erythrocyte of In Vivo Heat Exposed Rats Fed on Altered Dietary Protein Level
English2735Sudhamay GhoshEnglish Ajay K. ChatterjeeEnglishThis study was carried out to determine the effect of altered dietary protein level on the antioxidant status of erythrocytes of in vivo heat exposed rats. Experiments were performed on adult (80-90 days old) male Wister strain rats, divided in six groups of five animals each. Animals were exposed to 43±1 °C, ambient humidity conditions for 3 h daily for 15 days in a well maintained climatic chamber. Increased TBARS productions were found when measured in packed erythrocytes and erythrocyte membranes in both the rats fed on 18% or 6% protein diet after acute and chronic heat exposure. Supplementation of ascorbic acid to the rats fed on 18% or 6% protein diet was found to reduce the TBARS production remarkably in packed erythrocytes while it further raised the TBARS production in erythrocyte membrane in response to chronic and acute heat exposure. The liberation of alanine increased in 18% protein-fed as well as 6% protein-fed rats following chronic and acute exposure to heat. Decreased GSH content and increased membrane total thiol (-SH) content of erythrocytes were observed in both 18% protein-fed and 6% protein-fed rats. Prior supplementation of ascorbic acid was found to restore partially the GSH content of erythrocyte and to potentiate the increase in membrane total thiol (–SH) content in both adequately protein-fed and protein-restricted groups of rats.
EnglishAscorbic acid, Erythrocyte, Dietary protein, Heat stress, RatIntroduction
Red cells are always under high oxygen pressure and extremely susceptible to peroxidation and the conditions that favour peroxidation, are seemingly optimal in red cells [1]. Due to their function in carrying oxygen and their high iron content, erythrocytes are constantly exposed to oxidative stress [2]. Bernabucci et al. suggested that erythrocytes are an appropriate and sensitive model to study the oxidative status of transition dairy cows exposed to hot environment [3]. On the other hand, studies demonstrated that heat stress induces oxidative stress in the body [4, 5,6]. It has been demonstrated that heat stress is one of the most important stressors in the hot regions of the world(Altan, 2003).It results the generation of free radicals and other reactive oxygen species(ROS) (Chihuailaf et al., 2002) which leads to cellular and tissue damage (Tkaczyk andVizek, 2007). Free radicals and ROS have been demonstrated to have adverse effects on erythrocytes [7,8,9,10].Oxidative stress has been implicated with increased protein degradation through several mechanisms, including the activation of proteases and increasing the expression of genes involved in autophagy and proteolysis [11, 12,13]. ROS have been shown to impair protein synthesis by preventing mRNA translation [14, 15, 16, 17,18].Acute heat-stressed broiler chickens had a 2-fold increase in malondialdehyde, a marker of lipid peroxidation, in skeletal muscle [19]. End-product of lipid peroxidation, acts as a heat shock protein (hsp) inducer [20]. Membrane cholesterol contents modify the protective power of flavonoids against oxidative stress in erythrocytes [21].The antioxidant power of flavonoids, in turn depend on the depletion or incorporation of cholesterol in the erythrocyte membrane [22]. Ascorbic Acid is an outstanding antioxidant found in the human blood plasma [23] and it stabilizes free radicals and terminates free radical induced lipid peroxidation of cytochromes, thereby maintaining the structural integrity of cells [10,24].Further, it is known that ascorbic acid along with electrolyte supplementation was found to ameliorate the heat stress in buffaloes and plays a key role in the modulation of glutathione oxidase-reeducates system [25].It is concluded that specific cytosolic proteins are trans located to the membrane in human erythrocytes exposed to heat stress and they may play a novel role as erythrocyte membrane protectors under stress by stabilizing the membrane skeleton through their interactions with skeletal proteins(Sharma S, Zingde SM, Gokhale SM., 2013)[26]. Again, it is reported that the responses to heat stress are not only dependent on denatured proteins. The membrane lipid changes and the associated production of signalling metabolites clearly have a pivotal function to modify the heat stress response [27].In addition, dietary protein level is also known to have profound influence on antioxidant system and membrane composition and stability[28]. Nutritional modifications may help animals to prevent nutrient deficiencies that result from heat stress. Lower food intake during hot weather reduces nutrients available for absorption, and absorbed nutrients are used less efficiently [28].
Accordingly, the aim of the present investigation is to study the effect in vivo of heat stress on biochemical changes of erythrocyte in terms of antioxidant status, lipid peroxidation and glutathione metabolism on dietary protein adequacy and inadequacy. It is also intended to note whether ascorbic acid supplementation has any protective effects against chronic and acutely heat-induced membrane changes.
Materials and methods
Chemicals
Bovine serum albumin (BSA), 5, 5/-dithiobistrinitrobenzoic acid (DTNB), Folin-Ciocalteau reagent, tris, disodium ethylene tetra acetate (EDTA), reduced glutathione (GSH), were purchased from SRL, Mumbai, India. 5-Sulphosalicylic acid dihydrate and 2-thiobarbituric acid (TBA) were purchased from Spectrochem, Mumbai, India. Other chemicals used throughout the investigation were of analytical grade.
Animals and diets
Male growing rats of Wistar strain weighing 100-120g were used for the present study. The animals were kept in a well- ventilated room with 12 hrs. day - light cycle. The animals were accustomed with this condition for 7 days with adequate amount of food containing protein (casein)18%, carbohydrate(amylum) 71%, fat(groundnut oil) 7%, salt mixture 4% and adequate amount of vitamins mixture as reported elsewhere [29]. The composition of the salt mixture used was as described by Hawk and Oser [30].Then the animals were divided into four groups of equal average body weight. The animals of half of the groups were continued with diet containing 18% protein while those of the remaining groups were maintained on the diet containing 6% protein, and 83% carbohydrate. The 18% protein was used as it was considered as an adequate (normal) dietary protein level which was used on earlier occasions [29]. The 6% protein was used as an inadequate dietary protein level (protein inadequacy) to study the influence of dietary protein inadequacy. This experiment was approved by the guidelines of Institutional Animal Ethics Committee of department of Physiology, University of Calcutta.
Heat Exposure
After maintaining for three weeks on experimental diets, rats of experimental groups were exposed to heat stress. From one week before the onset of heat exposure, body weight, food intake and rectal temperature had been recorded on every alternate day till the termination of the heat-exposure period. Rats were exposed to heat stress in a well maintained climatic chamber.
Ascorbic acid was supplemented to the rats at a dose of 20mg per 100g of body weight intraperitoneally from one week before the beginning of heat exposure to increase the antioxidant reserves of the rats, and supplementation was continued till the day before their sacrifice. Effective thermal stress was determined by varying the duration of heat exposure and keeping the exposure temperature constant and vice versa. In both conditions rectal temperature of each rat was recorded at regular time intervals and also after the termination of heat-exposure in each day to know the pattern and degree of heat stress imposed on the rats. Following these approaches temperature of 4310C with 2 hrs. duration per day for 15 successive days and 4310C temperature with 3 hrs. duration in one day were considered optimum to produce the effect of chronic and acute heat exposure, respectively. Rats were exposed to heat between 2 and 6 p.m. in each day to avoid the diurnal variation of temperature. To maintain the uniformity in the heat stress induced, no experiment was performed in the months of summer (April to June) and winter (December to February). The entire study was carried out with several sets of experiments involving different groups of rats and keeping all the above conditions identical.
Tissue Collection
At the end of experimental period rats were kept fasting for 18 hrs. and then sacrificed by cervical dislocation. Blood was collected immediately from the hepatic vein with a heparinized syringe and kept in polypropylene vials at 40C, taking proper care to prevent any chance of haemolysis. To obtain erythrocytes, heparinized blood was centrifuged (1000xg at 40C for 10 mins.). Plasma was collected and stored in deep freeze. The buffy layer was removed completely by aspiration. The erythrocytes were washed three times with 20mM Tris-buffered-saline solution. The washed and packed erythrocytes were used for the preparation of ghost membrane.
Preparation of Erythrocyte Membrane
Fresh blood obtained from the rat was used to prepare ghost membrane following the procedure as developed by Marchesi and Pallade [31]. The packed red cells were washed thrice in 130mM NaCl and 20mM Tris-HCl (pH 7.4) mixture and recovered by centrifugation at 2500 r.p.m. for 15mins. Saline-washed red cells were lysed in 5mM Tris-HCl (pH 7.4) buffer containing 1mM EDTA and kept at 40C for 15 mins. Thirty nine volumes of haemolysing fluid were added to 1 volume of packed red cell, followed by centrifugation at 25,000g for 30 mins. at 40C. The supernatant was discarded and the membrane settled down was resuspended in the same medium and centrifuged again. The same procedure was repeated for 3 to 4 times until the membrane became milky white. Finally, the membrane was suspended in 50mM Tris-HCl buffer (pH 7.4) containing 1mM EDTA and kept frozen. All operations were carried out in cold.
Estimation of Membrane Protein
The protein was estimated by modified Lowry method [32]. The absorbance was read at 500nm and the protein content was calculated using a standard curve prepared with aliquots of BSA-solution having known concentration.
Statistical analysis
The data obtained from each experiment described above (NEnglishhttp://ijcrr.com/abstract.php?article_id=2289http://ijcrr.com/article_html.php?did=22891. Çimen MYB. Free radical metabolism in human erythrocytes. Clinica Chimica Acta 2008; 390 (1-2): 1-11.
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[18] Pietrocola F, Izzo V, Niso-Santano M, Vacchelli E, Galluzzi L, Maiuri MC et al. Regulation of autophagy by stress- responsive transcription factors. Semin Cancer Biol 2013 23: 310-322.
[19] Mujahid A, Akiba Y, Toyomizu M, Olive oil-supplemented diet alleviatesacute heat stress-induced mitochondrial ROS production in chicken skeletal muscle. Am J Physiol Regul Integr Comp Physiol 2009; 297: 690-698.
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[22] José I, Sánchez-Gallego, López-Revuelta A, José L, Sardina Hernández Hernández A, Sánchez-Yagüe J, Llanillo M. Membrane cholesterol contents modify the protective effects of quercetin and rutin on integrity and cellular viability in oxidized erythrocytes. Free Rad Biol Med. 2010; 48(10) : 1444-1454.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30HealthcareProfiling of Body Hydration and Related Blood Parameters under Hot-Arid Conditions among Indian Population doing Graded Physical Activity
English3642Abhishek BharadwajEnglish T. P. BaburajEnglish Amir Chand BajajEnglish Gulab SinghEnglish Usha PanjwaniEnglish Shashi Bala SinghEnglishThis study highlights the effect of extreme heat stress exposure (WBGT Index 36.24°C) on hematological variables while performing graded physical activity (sedentary, moderate and heavy). Total 30 participants were involved in the study and were randomly distributed into batches of 10 under three categories of physical activity. Volunteers performed physically strenuous work while being exposed to ambient heat stress (hot –arid conditions). White Blood Corpuscles (WBC) count in heavy work group [506 kcal/ hr energy expenditure] showed significant increase from pre to post exposure (p < 0.01). No other group showed significant change in WBC count. Haematocrit % (Hct %) was significantly increased among heavy work group (p < 0.01) whereas Red Blood Corpuscles (RBC) count and Red Blood Cell Distribution Width (RDW) showed no significant increase. Blood viscosity showed increase among moderate work group [412kcal / hr energy expenditure] (pEnglishHeat Stress, Graded work, Hypo-hydration, Blood parameters
Introduction
Tropical latitudes corroborate with warm climatic conditions ranging from arid / semi-arid terrain of desert to hot humid coastal regions. Soaring ambient temperature with intense solar radiation, warm winds and sandy terrain are salient features of desert environment. Such climatic conditions challenge the natives and also people carrying out operations (physically strenuous activities) under such environment. Desert conditions are known to cause debilitating effects on humans (1) with significant reduction in performance (9).
For maintaining a homoeothermic condition, human body toils hard to sustain thermal equilibrium (17). Result of heat exposure varies from mild discomfort, declined physical / mental ability and in extreme conditions threatening physiological conditions leading to mortality. Scarcity of potable water adds to the woe leading to altered body hydration status. During heat wave significant mortality and morbidity occurs due to heat stress and dehydration (5). Dehydration is primarily attributed to inadequate water intake caused by dysfunction of the central nervous system mechanisms controlling thirst (5). Symptoms of acute dehydration vary with the degree of water deficit. It is an established fact that hypo-hydration can lead to haemo-concentration. Reduced thirst and ingestive behavior have been reported in response to hyper-osmotic stimuli, hypovolemic stimuli and dehydration (5). Sufficient body water is required to sustain thermoregulatory function, thus losses in total body water can challenge the thermoregulatory system. A total body water deficit ?2% body mass (hypohydration) is recognized as the threshold when thermoregulatory function becomes measurably altered (19, 20). Sensation of thirst with dryness of mouth is felt at approximately 3%. The threshold for impaired exercise thermoregulation is 1% dehydration, and at 4%, decrements of 20 – 30% are seen in work capacity (8). Hypohydration decreases cutaneous vasodilation and sweating during heat stress (30, 31). Hypovolemia and hyperosmolality due to thermal dehydration also suppress thermoregulatory responses of sweating and cutaneous vasodilation, resulting in increased risk of heat illness (18). It has been often observed that non-acclimatized individuals tend to consume inadequate fluid leading to hypohydration, which can quickly result in dehydration with increased risk of heat illness / injury (27). Heat acclimatization induced effective adaptations are more pronounced after fluid-regulatory strain from permissive dehydration (11). In response to such altered hydration status, body undergoes several physiological changes. Exercise performed under such condition imposes severe strain on cardio-vascular system with finite cardiac output (CO) having to cater for increased blood flow to skin for thermal regulation (19, 28). Secretion and evaporation of sweat is important for thermoregulation but without water replenishment (hypohydration) plasma volume decreases with increased plasma osmotic pressure (hyperosmotic hypovolemia). Both osmotic and / or volume stressors delay onset and / or reduce the sensitivity of sweating and blood flow responses (19). It was also observed from study by Harrison et al (14) that exercise performed in heat reduced blood volume exacerbating strain on cardiovascular system. Altered hydration status affects the homeostasis within tissues and organs resulting in loss of plasma volume (haemo-concentration). It was further observed by Patterson et al that plasma volume was not defended more vigorously following humid-heat acclimatization. Rather a greater fluid loss may well underlie the mechanisms for enhancing plasma volume recovery when heat acclimation is induced using controlled- hyperthermia (26, 27). Sufficient body water is required to sustain thermoregulatory function, thus losses in total body water can challenge the thermoregulatory system (18). This situation aggravates further with exposure to high temperature and physical work under such conditions (9). Hypohydration may occur from voluntary fluid restriction, insufficient fluid availability or thermoregulatory sweating (18).
From the aforementioned findings, it can be deduced that strenuous activity under extensive environmental stress would lead to changes in body fluids and hematological parameters, ultimately affecting physical performance of the individual. Several hypotheses exist with regard to the physiological mechanisms responsible for heamodilution or haemoconcentration during heat exposure. Plasma volume expansion has been attributed by many to an influx of protein into the vascular volume, although some researchers attribute the same to electrolyte shifts. Though mechanism of action still remains speculative, it appears hematological parameters and plasma are crucial haemodynamic changes associated with adjustments to heat exposure (29).
Present study was carried out with two intentions:
To characterize hematological responses under high ambient temperature. These finding may be beneficial to devise strategies for identifying hypo-hydration status much before their symptomatic expression.
To document the effects of extreme heat stress exposure (WBGT Index 36.24°C) on hematological variables, while performing incremental graded work.
Materials and Methods:
Volunteers:
Thirty [30] participants were selected randomly without bias. They were physiologically and psychologically sound having mean age of 24 (±3.8) years, height 171 (±4.5) cm, weight 63.5 (±6.6) kg and BSA of 1.73 (±0.09) m2. The volunteers were divided into three (03) groups depending on nature of work (energy expenditure) they indulged in, viz. sedentary, moderate and heavy (8). Energy expenditure was measured during physical activity of the participant using “Actical Physical Monitor” (Actical Ver. 2.02 Mini Meter Company Inc. Bend, Oregon, USA). Sedentary group had energy expenditure of less than 3.0 Mets whereas moderate and heavy work group had Mets of 5.72 and 7.02 respectively (2). Prior to experiment, all participants provided their written informed consent, indicating they understood all procedures, risks and benefits of the study and their participation was voluntary. Testing protocol was approved by the Institutional Ethical Committee (IEC).
Experimental protocol: General:
The evening before day of exposure, participants for the study were briefed about procedure of experiment and their informed written voluntary consent was obtained. Caution was taken while briefing so as not to affect the outcome of experiments. Participants were asked to report the following morning before starting their daily routine. On the morning of exposure day, basal physiological readings were noted and blood sample drawn. Pre-exposure nude body weight was recorded [ATCO Human Weighing Balance, Mumbai, India]. Heart Rate was recorded using “Pulse Oxymeter” [Pulse Oxymeter Nellcor, USA. Model No.: N-20SA], prior to and also during exposure. Participants were issued “Actical Physical Monitor” for measurement of ‘Energy Expenditure’ during the period of exposure (Actical Ver. 2.02 Mini Mitter Company Inc. Bend, Oregon, USA).
Meteorological Parameters:
Environmental variables viz. Dry Bulb (DB) Temperature; Wet Bulb (WB) Temperature [Assmann Psychrometer THEIS, Gottingen, Germany] and Globe Temperature (GT) [measured manually using a Black Globe Thermometer] were monitored during entire exposure, along with Wind Velocity [Electronic Anaemometer, ACD Machine Control Co. Mumbai].
Black Globe was left out in open where volunteers were being exposed for determining solar heat load (radiation heat). Data from DB and WB were put onto a ‘Psychometric Chart’ for determining relative humidity (RH). From recorded data “WBGT Index” was calculated. “Solar Radiation” was also considered while computing the WBGT Index (16).
Physiological Parameters:
Blood Sampling:
All participants reported to laboratory on the morning of experiment day, before starting their assigned duties. They were asked to lie down in supine position comfortably for 5 minutes before drawing blood. A tourniquet was applied lightly and for minimum duration. With individual still in supine position venous blood was drawn and stored into vaccutainers (pretreated with EDTA K3). On conclusion of assigned duties done under heat exposure, post exposure blood samples were drawn from the same participants. Precaution was taken to maintain similar position and procedure as pre exposure sampling.
Blood Parameter Measurement:
Hematological parameters as Red Blood Corpuscles (RBC) count, White Blood Corpuscles (WBC) count, Red Blood Cell Distribution Width (RDW) and Haematocrit (Hct) % were measured. Blood parameters were measured using fully automatic ‘Hematological Cell Counter’ [MS4 Hematological Cell Counter. Melet Schloesing Lab, France]. Fresh, whole blood was used for assay. Whole blood was collected in vaccutainers pretreated with EDTA K3. After homogenization of the vaccutainers over roller, 20μl blood was aspirated by the instrument and used for assay. Assay was carried out on pre and post-exposure blood samples.
Viscosity Measurement:
Viscosity of pre and post exposure blood samples was measured from fresh whole blood using Micro viscometer (Model Cap 200L. Brookfield Engineering Laboratory, Brook Field USA). Estimation was carried out using 40 μl of blood.
Hydration Status:
During exposure a diary was maintained for noting fluid intake and urine output. Intake of fluid was documented by measuring amount of fluid consumption by the individual at every instance with help of graduated tumblers (provided to the participants on the day of experiment). Similarly, urine excretion during that period was collected in graduated jerrican. This amount of fluid intake and urine output was noted during the time of exposure while doing work. Data from water consumed, urine excreted and difference between pre and post nude body weights were used to compute sweat loss and sweat rate (15), leading to estimation of actual body hydration status of participant.
Urine and Plasma Osmolality:
To establish hydration status in different working groups’ urine osmolality was measured along with plasma osmolality. Urine sample was collected during the activity period in jerricans and osmolality measured. This was correlated with serum osmolality, which was separated from whole blood sample taken from the volunteers. Urine and serum osmolality was assayed using freezing point depression osmometry.
Statistical Analysis:
Different activity groups were compared by using One-Way ANOVA followed by Students Newman Keuls (SNK) Multiple Range Test. Data were considered significant with p < 0.05.
Result:
Meteorological Parameters:
Meteorological data are essential for assessment of environmental heat load. Physical data were collected from DB, WB, GT and wind velocity to assess intensity of prevailing heat stress. WBGT (Wet Bulb Globe Temperature) Index was computed using these data evaluated to identify the environmental work / heat load on participants during exposure. The maximum ambient temperature recorded during exposure was 42.4°C (DB), Globe temperature 57.8°C and Relative Humidity 38%. WBGT computed was 36.2°C. Maximum wind velocity (Gusting) was recorded at 12.8 m/s. [Table 1]
Haematological Parameters: Pre Vs Post Exposure:
White Blood Corpuscles Count [WBC]
Difference was noticed in all categories of activity. Only significant difference was in ‘Heavy’ work group (pEnglishhttp://ijcrr.com/abstract.php?article_id=2290http://ijcrr.com/article_html.php?did=2290
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30HealthcareChlorpyrifos Impairs the Contraction of Uterine Smooth Muscle by Inhibiting the Secretion of Estradiol from Ovary in Rat
English4348Kamalesh DasEnglish Kaushik SarkarEnglish Partha Pratim NathEnglish Mukti MondalEnglish Goutam PaulEnglishObjectives: Chlorpyrifos (CPF) has been used heedlessly in India to protect crop plants from pest induced crop loss. The objective of the study was to examine the effect of CPF, a broad spectrum organophosphate pesticide, on the contraction of uterine smooth muscle in rat model.
Methodology: Two graded doses of CPF were administered orally at a dose of 5.4 mg/kg.BW /d and 8.1mg/kg.BW /d to female Charles foster rats for 30 days. The record of the movement of the uterus was obtained with isotonic transducer coupled to RMS Polyrite D. The serum levels of estradiol and progesterone were measured as per standard protocols of kits of ERBA Diagnostic Gmbh Mannheim Germany.
Results: We have observed a significant inhibition of the force and increase in frequency of the contraction of uterus in all phases of an estrous cycle dose dependently in CPF exposed groups of rats compared to control groups of rats. We have also observed significant decrease in the level of estradiol in a dose dependent manner in all test groups of rats. These results indicate that CPF impairs the contraction of the uterine smooth muscle probably by inhibiting the secretion of estradiol from ovarian follicles. Further, the oxytocin induced potentiation of the tonic tension of uterus was significantly counteracted by CPF. This result proves that CPF probably impairs the contraction of uterine smooth muscle by inhibiting the estradiol.
Conclusion: In conclusion, CPF impairs the contraction of uterine smooth muscles by inhibiting the secretion of estradiol from ovary in rat.
EnglishOvarian follicle, Tonic tension of uterus, Estrous cycle, OxytocinIntroduction
In modern agriculture, agrochemicals like organophosphorous pesticide plays an important role in increasing the food grain production (Gulati, 2015). Chlorpyrifos (CPF) is a group of organophosphorous pesticide widely used in agricultural fields, food processing plants, residential pest control, paint industry and veterinary practice (Gulati, 2015). CPF has been detected considerable amount in fruits, vegetables, grains, dairy products, meat, fishes, soft drinks and also detected in cervical fluid, cord blood, breast milk and meconium (Angioni, 2011; Feei, 2008; Mathur;2003, Bedi, 2013 and Ostrea, 2006). The sources of human exposure to CPF are the food intake, inhalation and skin penetration during occupational use (Reigart, 2013). Within the human body, CPF is converted into chlorpyrifos oxon (CPO) in liver by cytochrome P450 mediated desulfuration and subsequently hydrolyzed to diethyl phosphate and 3, 5, 6- trichloro -2-pyridinol (TCPY) (Hodgson, 2008). It has been reported that CPF induces oxidative stress, inhibits the activity of AChE, promotes neurotoxicity and degeneration of dopaminergic and cholinergic neurones (Kalender, 2012; Slotkin, 2006; Whitney, 2012; Lee, 2012). CPF also exerts reproductive and developmental neurotoxicity in animals due to its endocrine disrupting function (Birnbaum, 2012; Farag, 2003; Flaskos, 2012). The reproductive effects of CPF in mammalian animals have been reported discretely. It has also been reported that CPF causes disruption of the estrous cycle rhythmicity, impairment of the growth of primordial follicles in the ovary and disruption of the of breast cell cycle through production of reactive oxygen species (oxidative stress) (Kamalesh, 2014; Ventura, 2012). But the effect of CPF on the function of uterus, one of the major female reproductive organ, has not been reported till date. It provides the habitat for implantation and growth of the fetus till maturation. Besides, it helps in the parturition of the fetus through its vigorous contractions. Histologically the wall of the uterus contains three layers from inside to outside; endometrium, myometrium and perimetrium. The myometrium of uterus is composed of visceral smooth muscle cells which are organized in circularly and longitudinally. These visceral smooth muscle cells provide rhythmic contraction of the uterus in the basal state due to their spontaneous rhythmic excitation intrinsically (Khan, 2001). The basal contraction of the uterine smooth muscle cells are regulated by oxytocin released from the posterior pituitary. Estradiol released from the developing follicle of the ovary potentiates and the progesterone released from corpus luteum inhibits the oxytocin induced contraction of uterine smooth muscle (Jackson, 1998; Fanseca, 2003). Several reproductive functions including the transport of sperms and embryo, menstruation, pregnancy and parturition will be impaired if the basal uterine contractions are changed due to alterations in the levels of oxytocin and ovarian estradiol and progesterone. Considering the toxic effects of CPF on the functions of several reproductive organs, we predict that CPF might exert harmful effects on the function of uterine smooth muscle cells. So, the objective of the present study was to examine the probable effects of CPF on the contractions of uterine smooth muscle in rat.
Materials and Methods
Chemicals and reagents
All the reagents used this study were of analytical grade. Sodium chloride (NaCl), Potassium chloride (KCl), Calcium chloride (cacl2), Sodium bicarbonate (NaHCO3), Magnesium chloride (MgCl2), Sodium dihydrogen phosphate (NaH2PO4), and Glucose were procured from E. Merck, India. Chlorpyrifos (CPF) was purchased from Devidayal (sales) Limited, Mumbai, India.
Oxytocin was purchased from E. Merck, India.
Animals
Adult female Charles foster rats aged 16-18 weeks and weighing 120-160gm were selected for the experiment. The animals were matched with age and by body weight across the treatment groups at the start of the experiment. Rats were housed in polypropylene coated plastic cage containing paddy husk bedding. Polypropylene cages contained female Charles foster rats, were kept in the animal house and maintained at an average temperature of 27• ± 1• C with 12:12h light and dark cycle (light on around 8-9 A.M).
Ethical approval
The rats were kept in the Molecular Neurotoxicology laboratory, Department of Physiology, University of Kalyani and were maintained as per recommendation of the animal ethical committee of University of Kalyani in accordance with the national guidelines.
Experimental setup
Animals showing normal and regular estrous cycle (4 to 5 days) for at least three consecutive times were selected for the study. After 10 days of acclimatization, the rats were distributed to three groups. The rats of the first group were received standard laboratory diet and were designated as control group. The rats of the second and third groups were received 5.4mg/BW/d and 8.1 mg/BW/d chlorpyrifos for 30 days duration and were termed as treated groups. After completion of treatment, rats were sacrificed by cervical dislocation and blood was collected for the determination of estradiol and progesterone. For study of uterine movement, rat was sacrificed and abdomen was immediately opened and the uterine segment of 4 cm was removed by transverse incision. Estrous cycle study (Bancroft, 2002) was performed to identify the specific phase on the day of experiment.
Uterine motility study
Devices: Glass jar Bath
A glass water bath of about 750 mL capacity fitted into a metal stand in which a fixed heater was located to maintain the temperature as required. An inner glass tube (Organ bath) of 40 mL capacity passed through the bottom of the stand and was connected by a T-shaped glass tube. Preparation of rat’s uterus
The mature female Charles foster rats at different phases of estrous cycle (i.e. proestrus, estrus, metestrus and diestrus) were used where vaginal lavage was performed for detection of the stages of sex cycle of the animals. Animals after being examined were sacrificed and the uterus was dissected out and luminal content was gently flushed out. Uterine horn was suspended in the organ bath containing warm oxygenated (95% O2 and 5% CO2) physiological salt solution (Nacl 8.0g/l, KCl 0.20g/l, CaCl2 0.2g/l, MgCl2 0.1 g/l, NaHCO3 1.0 g/l, NaH2PO4 0.05 g/l, Glucose 1 g/l, PH 7.4) at 37±0.5•C. One part of each uterine strip was connected with glass hook placed at the bottom of the organ bath and another part was attached to the isotonic transducer using metal thread.
Recording of uterine motility using RMS Polyrite D and Isotonic transducer
The uterine movement was achieved with isotonic transducer (IT-2245) coupled with RMS Polyrite D, Software (RMS, INDIA).
Hormonal study
Levels of serum 17-β estradiol (E2) and progesterone (P4) were estimated by enzyme-linked immunosorbent assay (ELISA) methods using kits of ERBA Diagnostic GmbH Mannheim Germany.
Statistical analysis
Uterine activity was characterized by episode of uterine contractions separated by periods of silence. All the data were expressed as mean± SEM of the value of each experimental group. Force of contraction was measured in terms of amplitude, frequency and duration. Statistical comparisons between the values obtained in control and in treated rats were carried out by using a Student’s t test for paired group. The number of the uterine preparation used in each experiment is indicated by the alphabet ‘n’ in the result. P value Englishhttp://ijcrr.com/abstract.php?article_id=2291http://ijcrr.com/article_html.php?did=2291
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Kalender Y, Kaya S, Durak D, Uzun FG, Demir F. Protective effect of catechin and quercetin on antioxidant status, lipid peroxidation and testis histoarchitecture induced by Chlorpyrifos in male rats. Environ Toxicol Pharmacol 2012; 33(2): 141-48.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30HealthcareAlterations in Carotid Body Morphology and Cellular Mechanism Under the Influence of Intermittent Hypoxia
English4957Utkarsha KumarEnglish Dishari GhoshEnglish Snigdha ShawEnglish Gopinath BhaumikEnglish Rajinder K GuptaEnglish Prasanna K ReddyEnglish Shashi Bala SinghEnglishAims: Intermittent hypoxia (IH) training is said to have a preconditioning effect for evoking acclimatization at high altitude (HA). Carotid body (CB) plays a vital role in oxygen sensing and is an important component in HA acclimatization. The present study reports the mechanistic effects of IH that involves episodes of hypoxia of few hours continued for several days, on the CB responses to acute hypobaric hypoxia in terms of morphological changes in CB and its cellular functions.
Methodology: 24 Sprague-Dawley (250-300g) rats were divided into 2 major groups: 1) control, 2) experimental group (n=12 each) in which the rats were exposed to IH training for 10 days with a single hypoxic episode of 4h/day at a simulated altitude of 15000ft. 6 rats from each group were further subjected to a simulated hypobaric acute hypoxic (AH) challenge of 1hr at 25000ft to see the effect of IH training (IHT) and were named as 3) Control+ AH challenge and 4) IHT+ AH challenge. Morphological changes in CB in different groups were observed along with expression of hypoxia inducible factor (HIF) 1α, HIF2α, NADPH Oxidase 2 (NOX2) and Superoxide Dismutase 2 (SOD2) using immunohistochemistry for the first time.
Results: The results showed that IH training leads to morphological changes in terms of hyperplasia and unaltered HIF1α levels along with a highly significant rise in HIF2α in CB. When the rats are exposed to AH without IH conditioning, there is a significant rise in HIF1α and thus NOX2 levels. However, prior exposure to IH leads to a significant rise in the HIF2α levels and thus SOD2 levels, when subjected to AH challenge.
Discussion and Conclusion: These results indicate that IH training affects the cellular response of CB by regulating balanced expression of both HIF1α and HIF2α, thus modulating the cellular redox state by promoting the antioxidant enzyme production and suppressing the pro-oxidant enzyme levels, thereby playing a crucial role in pre-conditioning to acute hypoxia.
EnglishCarotid Body, Intermittent hypoxia, HIF1?, HIF2?, NOX2, SOD2Introduction
Carotid bodies (CBs) are small neurovascular structures at the bifurcations of the common carotid arteries comprising of Type I (chief or glomus) and type II (sustentacular) cells [1] that recognizes the oxygen (O2) deprivation (hypoxia) in the arterial blood. Various morphological changes have been reported in mammalian CB on chronic as well as on acute hypoxic exposure. CBs protect the organs from hypoxic damage by releasing neurotransmitters (NTs) and instantaneously signaling the brainstem respiratory centre via the carotid body nerve, resulting in hyperventilation that is an integral part of altitude acclimatization [2-4]. Currently, models of oxygen sensing are based on either a heme protein or the production of reactive oxygen species (ROS) by NAD(P)H oxidases and mitochondria [5]. Other than these, HIF1 and HIF2, members of hypoxia inducible factor (HIF) family are believed to be chief molecular determinants for CB O2 sensing. They constitute a common HIFβ subunit with oxygen regulated HIF1α and HIF2α subunits respectively [6,7]. HIF1α is a potent activator of genes encoding pro-oxidant enzymes such as NADPH oxidase 2 (NOX2) [8] and HIF2α regulates antioxidant enzymes such as superoxide dismutase 2 (SOD2) [9]. The cellular redox state relies on the regulation of these enzymes by the respective HIFα isoforms that further triggers the signaling pathways such as the release of multiple NTs [10,11].
Hypoxia and intermittent hypoxia (or IH i.e. repeated episodes of hypoxia interspersed with normoxic or less severe hypoxic episodes) mediate oxygen sensing in the CBs. It has been well documented that IH as in obstructive sleep apnea (OSA) cases (hypoxia for 10-40 seconds, normoxia of various minutes) [10] showed no significant effect on the morphology of CB, be it CB hypertrophy or hyperplasia of glomus cell volume, though it increases CB’s sensitivity and toxic sympathetic activation [12]. Moreover, studies reveal that augmented activity of the CB, induced by IH plays an important role in the pathogenesis of sleep apnea [13] in anesthetized rats and humans and emphasis is being made to understand its underlying mechanisms. Furthermore, the equilibrium between HIF1α and HIF2α, hampered by chronic IH leads to an imbalance between pro-oxidant and antioxidant enzymes respectively that causes oxidative stress resulting in pathophysiology [14-16] and has been implicated as a risk factor for an array of cardiovascular diseases [17] affecting humans.
IH training (IHT) is being extensively used by sports medicine community to improve performance and by the US army to induce pre-acclimatization without any pharmacological interventions [18]. A low number of re-oxygenation cycles per day and moderate hypoxic episodes elicits the beneficial effects beyond pathology and provides protection against hypertension, myocardial injuries, heart arrhythmia and bronchial asthma [19,20]. It helps in activating the surviving mechanisms against acute hypoxic exposure by increasing haemoglobin, erythropoietin production, total red blood cells, exercise time, hypoxic ventilatory responses in healthy human subjects, bone mineral density, respiratory and non respiratory somatic motor recovery following spinal injuries in rats and humans and enhancement of spatial learning and memory without any detectable adverse consequences [21].
The effect of the paradigm of IH resembling recurrent apneas and the role of CB in the disease mechanisms has been studied considerably in the recent years. Also, studies have been focused upon the IHT consisting of hypoxic episodes of few hours in human subjects. However despite these aforesaid extensive studies, the pre-conditioning effect of IH of long duration on the rat CB and the probable cellular mechanism whereby IH mediates the CB activity remains unstudied. In the present study, we evaluate the mechanistic effects of IH on CB responses to acute hypoxia in rats, in terms of changes in CB morphology and cellular functions.
Materials and Methods
Animals
Male Sprague-Dawley rats (250-300g) were used for all experiments that were maintained under a 12-h light–dark cycle at temperature 25±1°C, humidity 55% ± 2%, in the Institute’s animal house facility. The study was approved by the Animal Ethical Committee of the institute in accordance with Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA) of the Government of India. All efforts were made to minimize the animal suffering and to reduce the number of the animals used.
Experimental Design and Hypobaric Hypoxia Exposure
Animals were randomly assigned to two main groups:
Control (n=12) rats maintained in normoxic condition.
Experimental group (n=12) rats subjected to intermittent hypobaric hypoxia (IH) with hypoxic episode of 4h/day at 15000ft for 10 days [13] (with modifications) in animal decompression chamber. The airflow in the chamber was 2L/min with relative humidity 40%-50% and temperature 25±1?C.
6 rats from each of these groups were exposed to an acute hypoxic challenge (AH) at simulated altitude of 25000ft for 1h to assess the effect of IH conditioning on acute hypobaric hypoxic exposure and to see whether the type of IH modality including longer durations of hypoxic exposure has some beneficial role against the deleterious effects of hypoxia or not. The groups were named as Control+ Challenge (C+AH) and IH+ Challenge (IHT+AH) respectively.
General preparations of the animals and tissue
After the hypoxic exposure, rats were anaesthetized with ketamine/xylazine (70 and 6mg/kg, i.p., respectively). An anterior neck incision was made and various neck muscles were retracted to expose carotid bifurcation along with CB. The tissue was perfused transcardially first with 0.05M phosphate buffer saline (PBS), followed by 4% paraformaldehyde in O.1M phosphate buffer, pH 7.3. After perfusion, the CBs were immediately excised and were post-fixed in the same fixative overnight at 4?C.
Histological Studies
The fixed tissues were dehydrated, cleared and embedded in paraffin blocks and 5-6 µm thick tissue sections were obtained using a rotary microtome that were stained with haematoxylin and eosin. The sections were examined under the light microscope.
Immunohistochemical Studies
The sections were dewaxed with xylene and rehydrated with a series of decreasing grade of ethanol solutions. They were then immersed in antigen retrieval solution (0.1M citric acid buffer, pH 6.0) for 10 min in water-bath (100?C) and to block endogenous peroxidase activity, they were immersed in 4% hydrogen peroxide for 5 min at room temperature (RT). Sections were pre-incubated (2h) with blocking serum to reduce non-specific binding of the antiserum. Between the separate steps, the sections were rinsed with cold PBS and incubated with the following primary antibodies: HIF1α (mouse monoclonal IgG antibody, 1:500 dilution, Sigma Life Science); SOD2 (rabbit polyclonal IgG antibody, 1:1500 dilution, Sigma Life Science); HIF2α (mouse monoclonal IgG antibody, 1:200 dilution, GeneTex) and NOX2 (rabbit polyclonal antibody, 1:200 dilution, PromoKine), in 0.05M Tris/HCl buffer, overnight at 4?C. They were washed three times in PBS and incubated (30 min) at RT with respective secondary antibody labeled with horseradish-peroxidase. Finally, sections were washed and the peroxidase was visualized by immersing in 0.05% diaminobenzidine (DAB) for 3-5 min, rinsed in distilled water and counterstained mildly with haematoxylin, dehydrated in graded alcohols, cleared in xylene, mounted with DPX solution and air-dried. Positive staining was indicated by a brown color.
Image acquisition
Histological and immunohistochemical images were acquired using a Leica DMR (Germany) microscope via an inbuilt CCD Color Camera (Leica) with 40X objective lens.
Image analysis
The section with the largest area of cells was chosen to perform measurements to evaluate the following parameters:
Morphometric analysis was performed with NIH Image J software program.
The total number of type I cells in the CB were counted using a mechanical cell counter in Image J with 40X objective lens.
Values of integrated density (product of Area and Mean Gray Value of the pixels in the image) of immunohistochemical images of different groups were obtained using Image J (selected using the freehand selection tool) in order to quantify the expression levels of HIF1α, NOX2, HIF2α and SOD2 by averaging their respective values.
Histogram profiles of the images were obtained by Image J by digital image analysis that is a plot of the intensity values of the pixels (x axis) vs. the number of pixels representing the intensities (y axis) [22]. The pixel intensity values for any color ranges from 0 to 255 (0 represents darkest color shade; 255 represents lightest color shade as standard). The profiling is done using the averaged values of the number of pixels in respective values of intensities of various cells in the images in a particular group. The increase or decrease in the intensities of DAB (brown color) is analyzed by observing the shift of the histogram of hypoxic exposed groups to the left or right of the histogram obtained in the control group.
Statistical analysis
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7. Yuan, G., Peng, Y.J., Reddy, V.D., Makarenko, V.V., Nanduri, J., Khan, S.A., Garcia, J.A., Kumar, G.K., Semenza, G.L., Prabhakar, N.R.: Mutual antagonism between hypoxia-inducible factors 1alpha and 2alpha regulates oxygen sensing and cardio-respiratory homeostasis. Proceedings of the National Academy of Sciences of the United States of America 110(19), E1788-1796 (2013). doi:10.1073/pnas.1305961110
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN2017August23HealthcareAwareness and Attitude Regarding Cataract Surgery Among Senile Cataract Cases Attending Camps
English5863RadhikaEnglish Biradar SunilEnglish Udgiri RekhaEnglish Bellary ChandrashekharEnglishContext: Study conducted in district of North Karnataka where majority of the population are illiterate and have poor access to treatment of cataract. People have different concepts regarding cataract
Aims: The present study is an attempt to know the knowledge, attitude and practices regarding cataract surgery
Settings and Design: All the camp cases screened during the period Dec-May 2012 from 6 neighboring villages for 6 months were included in the study. The data was collected by interview technique
Methods and Material: A Cross-sectional study was carried over a period of 6 months. The results were analysed using proportion and chi-square test.
Statistical analysis used: The results were analysed using proportion and chi-square test.
Results: In the present study, 81% were illiterates and 50% were unemployed. Majority of them (67%) were females. The knowledge about cataract related to, ‘at what age it occurs’ and how it occurs was found statistically significant with relation to education. Regarding reasons for not getting surgery done, 64(21.3%) cases cited lack of escort as the reason, 148(49.3%) cases managed with existing eyesight, 49(16.3%) cases cited economic reasons. The socioeconomic status versus preference of surgery with or without lens found to be significant (pEnglishKnowledge, Attitude, Practices, Cataract surgeryINTRODUCTION
Blindness is a major public health problem in India. It is estimated that there are 9 to 12 million people blind in India which amounts to about one-fourth of all the blind people worldwide.1 In India, 3.8 million people contribute to cataract blindness annually.2 Out of the 41.8% global blindness, 23.5% is contributed by India.3 A major proportion of total blindness in India (81%) is constituted by cataract blinds. Yorston4 has suggested that four barriers in limiting access to cataract surgery worldwide: Lack of knowledge, uncertainty over the quality of available services, transportation problems, and concerns over the cost of surgery. The present study was carried out to assess the knowledge, attitude and practices regarding cataract surgery.
Subjects and Methods:
All the camp cases screened during the period Dec-May 2012 from 6 neighbouring villages for 6 months were included in the study. This was a hospital-based, crosssectional descriptive, clinical study. Inclusion criteria: consecutive patients, aged 30 years or above, with vision Englishhttp://ijcrr.com/abstract.php?article_id=2293http://ijcrr.com/article_html.php?did=22931. Thylefors B, Negrel AD, Pararajasegaram R, Dadzie KY. Global data on blindness. Bull World Health Organ 1995;73:115-21.
2. Minassian DC, Mehra V. 3.8 million blinded by cataract each year: Projection from the first epidemiological study of incidence of cataract blindness in India. Br J Ophthalmol 1990; 74: 341-9.
3. Thylefors B, Negrel AD, Pararajasegaram R, Dadzie KY. Global data on blindness. Comm Eye Health 1996; 9 (17): 1-8.
4. Yorston D. High-volume surgery in developing countries. Eye.2005;19:1083–9.
5. Dandona R, Dandona L, Srinivas M, Giridhar P, Vilas K, Prasad MN, et al . Blindness in the Indian state of Andhra Pradesh. Invest Ophthalmol Vis Sci 2001;42:908-16.
6. Gupta SK et al. Social implications of cataract related blindness in rural India. In: Kumar V, ed. Aging-Indian Perspective and Global Scenario. New Delhi: All India Institute of Medical Sciences. 1996; 126-9.
7. Melese M, Alemayehu W, Friedlander E, Courtright P. Indirect costs associated with accessing eye care services as a barrier to service use in Ethiopia. Trop Med Int Health 2004;9:426-31.
8. Dhaliwal U, Gupta SK. Barriers to the uptake of cataract surgery in patients presenting to a hospital. Indian J Ophthalmol 2007;55:133-6
9. Rabiu MM. Cataract blindness and barriers to uptake of cataract surgery in a rural community of northern Nigeria. Br J Ophthalmol 2001;85:776-80.
10. Rotchford AP, Rotchford KM, Mthethwa LP, Johnson GJ. Reasons for poor cataract surgery uptake - a qualitative study in rural South Africa. Trop Med Int Health2002;7:288-92.
11. Soundarssanane MB, Bansal RD, Narayan KA. An epidemiological study of cataract in a rural area of Pondicherry. Ind J Med Sci 1986; 40: 273-7.
12. Chatterjee A, Milton RC, Thyle Sydney. Prevalence and aetiology of cataract in Punjab. Br J Ophthal 1982; 66: 35-42.
13. J. Bhagwan, I.M. Rastogi, J.S. Malik, C.S. Dhull. Knowledge, Attitude and Practices Regarding Cataract Surgery among Senile Cataract Cases in Haryana .Ind J Community Medicine 2006;31,66-8.
14. Brilliant GE, Brilliant LB. Using social epidemiology to understand who stays blind and who gets operated for cataract in rural setting. Soc Sci Med 1985; 21: 553-8
15. Bowman RJC, Jatta B, Faal H, Bailey R, Foster A, Johnson GS. Long-term follow up of lid surgery for trichiasis in Gambia: Surgical success and patients perceptions. Eye 2000;14:864-8.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241916EnglishN-0001November30TechnologyFace Resolution Enhancement Using Artificial Bee Colony
English6470Kajal SharmaEnglishThe existing adaptive shock filter based algorithm generate resolution enhanced images with constant values and random number of iterations for all images. Here, a useful application of artificial bee colony to find the optimistic adjustment factor for DWT and adaptive shock filter for face resolution enhancement as well as super-resolution is implemented to find the solution to that problem. The guided image filter is used in order to preserve edges in a more optimistic manner. The importance of this paper is to consider the effect of noise and edge preservation.
EnglishDiscrete Wavelet Transform, Discrete Cosine Transform, Adaptive Shock Filter, Discrete Fourier Transform, Peak Signal Noise Ratio, Root Mean Square Error, Mean Square Error, Bit Error Rate, Mean Absolute Error, Artificial Bee Colony
Introduction
The principal objective of image enhancement isto enhance the understandability or view of data in images for human watchers. It is also helpful to give a better contribution for other automated image processing techniques. The graphic display for analysis become more accessible by accentuating or sharpening image features, for example, edges, boundaries, or contrast [1]. The inherent information contents of the data don't increase in the enhancement process. However, it expands the dynamic range of the picked features with the goal that they can be distinguished effortlessly. Nowadays, this has been turned into the concentration of the focus in numerous fields, for example, public security, archaeology, military, medical imaging and three-dimensional image [2]. The image enhancement techniques focus attention on certain features of interest in an image and draw out the features in an image that is obscured. The contrast adjustment, filtering, morphological filtering, and deblurring are the various enhancement techniques. A modified version of the original image is achieved through these image enhancement operations and can also be utilized as a preprocessing initiative to upgrade the after effects of image analysis techniques [3]. Frequency based domain image enhancement is a technique, which is utilized to give details of the study of mathematical functions or signals regarding frequency as well as operating them on the transform coefficients of the image, for example, discrete cosine transform (DCT) [4], Fourier transform, discrete wavelet transform (DWT). The motive of utilizing thisterm is to improve the image by controlling the transform coefficients. There are a few merits of Frequency based image enhancement such as including the low complexity of the calculations, facile vision as well as the easy applicability of special transformed domain properties, controlling the frequency composition of the image. The appearance of low-quality images is enhanced by working with traditional methods of image enhancement.
Super-resolution is a technique which is used to create a high-resolution image by taking one or various low-resolution images. This method is conceivable to describe an image or high-frequency components which may be missed because of numerous causes, ambient light from different elements, as well as the low number of camera sensory cells, the movement of the camera and not alter the camera's focal point.
The lost features of the image can be recovered by using this process of super-resolution. Also, super-resolution minimizes the blurring and aliasing of images and interpolates or predicts lost information from accessible evidences. In numerous applications of machine vision and image processing, the Super-resolution methods are used. Today, advancements came in systems; introducing higher precision and more processing power, which has made more consideration of super-resolution methods based on software. This technique is utilized for many monitoring machines, for example, face recognition, identification and license plate recognition, remote sensing, automatic target recognition, medical image processing, for example, CT and MRI and converting video to different standards (for example, converting NTSC to HDTV), image enhancing, processing of satellite images, microscopic image processing and astronomical image processing.
Adaptive Shock Filters
[Jinsheng Xiao et al.,] discussed that an adaptive shocking filtering model has following properties:
Fig 1: Flow chart of Image Super-Resolution and Enhancement
Artificial Bee Colony
In the Artificial Bee Colony model, there are three categories of honey bees: employed bees, onlookers and scouts. In this colony, this is supposed that there is merely one artificial employed bee for each and every food source. Also, the number of food sources in the model is equal to the number of employed bees around the hive. Firstly employed bees would go to their food source as well as after returning they dance on their hives. The food source of employed bee has become abandoned and turns into a scout and begins to look for searching another food source. The remaining group which is onlookers, watches the dances of employed bees as well as chooses food sources relying on their dances. Some of the key steps of the algorithm are given beneath:
1. For all employed bees, the initial food sources are produced.
2. Repeat the step until all employed bees get the food sources.
3. Then all employed bees remind food sources in their memory and go towards those food sources and afterward, a neighbor source determined, and then calculates the nectar amount and started dancing in the hive.
4. Onlookers choose and select their food sources by observing the dance of employed bees, and then go to that selected food source. She evaluates its nectar amount after choosing a neighbor around that.
5. When the scouts discover the new food sources, the abandoned food sources are resolved as well as substituted with those new food sources.
6. The elite food source is finalized from all of the available food sources.
7. UNTIL (requirements are met).
Discrete Wavelet Transform
The discrete wavelet transform is an execution of the wavelet transform utilizing a discrete set of the wavelet scales as well as translations complying with various characterized rules. Also, that type of transform disintegrates the signal into a mutually orthogonal set of wavelets. This transform is the primarydistinction from the CWT and its implementation of the discrete time series sometimes known discrete-time continuous wavelet transform. Similar to the case in Fourier analysis, the DWT is invertible, so that the initial signal can be totally recuperated from its DWT representation. Not at all like the DFT, the DWT, indeed, alludes not simply to a single transform, but instead a set of transforms, each with an alternate arrangement of wavelet basis functions. Haar wavelets and the Daubechies set of wavelets are two of the most common wavelets.
There is a scaling function from which the wavelet should be created and this scaling function portrays the scaling properties. There is a constraint that the scaling functions should be orthogonal to its discrete translations suggests various mathematical restrictions that are specified all around such as the dilation equation
Related Work
C. Munteanu et al. (2004) [16] has described a new approach to automatic Image enhancement. It is a visually more agreeable, low noise output to get the image as an input image. Therefore, image enhancement is a troublesome undertaking task to automate the procedure of analyzing and attempting to dispose of human intervention. This paper is another automatic image enhancement technology powered by a developmental adaptation process introduction. They have new objective criteria for enhancement, according to the test and try to find the best image. Due to the high complexity of criteria planned. They are the best for a global search strategy as an evolutionary algorithm (EA). They contrasted their technique with other automatic enhancement methods, such as contrast stretching and histogram equalization. The conclusions as far as both subjective and objective evolution demonstrates the excellency of their process.
Kabir et al. (2006) [17] has presented a technique for image contrast enhancement based on Block based intensity-pair distribution. Added intensity distribution utilizing the whole image, instead of the estimated intensity takes this image-pair algorithm proposed and an expanding distribution according to the intensity of the pixel map. A linear magnitude mapping function is utilized in place of non- linear to control over-enhancement and the contrast stretch. This preserves the gray level linear mapping.
Block local information easily contrast enhancement, intelligent edge brings out information and removes the noise from the image. For an extended analysis of image features, the approximate algorithm is the best.
Tokuda et al. (2007) [18] has defined thatgamma correction image quality adjustment parameter one of the image quality enhancement support system reflecting user subjectivity. Optical gamma value’s derivation as an optimization problem is studied in this. To reflect user subjectivity, a support system by Interactive Evolutionary Computing image quality enhancement is realized. This technology is a gamma value derived from manually, image quality and verified by comparison with etymology.
Jafar et al. (2007) [19] has proposed that contrast enhancement is an essential step in virtually every image processing and computer vision application to achieve the needed level of enrichment different applications for different methods. Consumer electronics such as any enhancement techniques to enhance both local and global details should be able, yet not at the expense of decay in the quality of the image. He called his early work on the advancement of new methods-based histogram equalization in the current multi-level component image contrast enhancement. This method is effective and simple and visible image at least distortion with both global and local contrast enhancement to allow thresholding the grey level histogram equalization method, multiple classic and connected component analysis taking advantage of the capabilities of Visual and quantitative evaluation technique for achieving this goal than revealed.
Chen et al. (2008) [20] has proposed a new fuzzy based contrast enhancement technique for remote sensing images.Traditional advancement methods frequently involve highly areas in black and bright local brightness details based on fuzzy set theory. Fuzzy set theory intended use grey due to the traditional division by values to avoid cracks. The main concept of the theory is that the elements of an interval [0, 1] instead of binary value membership degree. Hence, a complex process can be considered as an algorithm involves three stages: first, the satellite image gray-level c-fuzzy membership space is transformed by means of clustering. Secondly, each cluster of the suitable stretch model is constructed of the same membership base. Third, image gray-level space stretches back to the gray value changes by merger of each cluster. From the outcomes, obviously the, proposed technique can enhance the image conception and higher key measurement.
Lee. he et al. (2008) [21] has suggested that a new color image enhancement algorithm based on MSR. Appropriate wavelet base input image disintegrated in three levels to be selected. Then decompose the process input image into different enhancements algorithms employed in three levels. Then decompose the different enhancement algorithms Wavelet coefficients and coefficients were employed to scale. Wavelet coefficients, each level character, after a revised analysis of Adaptive non-fast mask technology, which improved image contrast, more clear and colorful image processing was used. Experiments show the projected algorithm enhanced some outdoor images obviously.
Gaps in Study
[Jinsheng Xiao, Guanlin Pang, et al.,] discussed that by conducting the review, it had been found that the various gaps in earlier work on image super-resolution and enhancement techniques.
1. The effect of noise and edge preservation is ignored in existing super-resolution and enhancement technique.
2. The use of artificial bee colony while super-resolution and enhancing the image is also ignored by existing researchers
3. The majority of existing research focuses on natural images. The effects of image super-resolution and enhancement on face images are ignored.
Methodology
PROPOSED ALGORITHM
1. Start by reading the image
2. Apply DWT on the image
3. It splits the image into four bands of data-LL, LH, HL, and HH.
4. Then apply adaptive shock filter on LL.
5. Afterward, apply artificial bee colony to find optimistic adjustment value.
6. Then adjust the actual image using optimistic artificial bee colony.
7. It will give the improved versions of all bands of data as seen in flow chart-Improved LL
8. Then apply inverse DWT.
Discussion
PSNR is calculated by taking three images. From the graph, it can be easily depicted that the PSNR using Artificial Bee Colony is more than that of PSNR using Adaptive Shock Filters. PSNRs in the case of proposed technique are 47.7104, 47.5603 and 47.8784.
Discussion
Mean square error is to figure an error signal by subtracting the test signal from the reference and afterward computing the average energy of the error signal. This can be calculated as:
Discussion
BER is the rate of occurrence of errors in a transmission system. It can be simplified as number of bit errors separated by the aggregate number of transferred bits during a considered time interval. This definition can be simplified into a basic equation:
BER is calculated using the proposed technique on three different images. From the graph, it is clear that the BER with proposed technique starts decreasing. The calculated Bit Error Rates for three images are 0.02096, 0.021026 and 0.020886.
Mean Absolute Error is calculated on three images using proposed algorithm. The achieved result is approximately half to theMAE with existing technique. MAEs with proposed technique are 1.9859, 1.7054 and 2.446.
Conclusions
This paper has tended to the issue of face super-resolution and enhancement in light of artificial bee colony. First, discrete wavelet transform is employed in the facial image which separates smooth variations and sharp variations. Then the adaptive shock filter is applied to various bands of data. Then, artificial bee colony is used to find optimistic adjustment values, which adjusts the actual image using optimistic ABC. This will give the improved bands of data. Finally, inverse discrete wavelet transform is employed in the image to reassemble the various classes of data into a reconstructed image. Finally, the guided image filter is applied to preserve edges in more optimistic manner and then an enhanced image along with super-resolution is reconstructed. The experimental outcomes prove that the proposed algorithm has preferable outcomes over the current technique based on adaptive shock filter model both in quantity and quality.
Acknowledgement
It is a great pleasure to express my deep sense of gratitude and heartfelt thanks to the various persons who have been of enormous help in one way or the other of successful completion of this research. They not only inculeated in me the fath and confidence but offered very tangible, moral, pyschosomatic support in every resect.
First and foremost I would like to express my gratitude to my thesis guide Prof. VikramMutneja for his guidance, advice and constant support throughout my research work. He has been a great source of motivation and always eager to help. It was an absolute pleasure to work under his guidance. Out of deep sense of gratefulness I express my sincere thanks to him. This research would have never acquired its existence without his help and cooperation. I also want to express my respects to Dr. Rajni, HOD of Electonics and communication Engineering Department.
I would like to thank all my family members and staff of the department of Electonics and communication Engineering, SBSSTC for their generous help in various ways for the completion of this research.
Least butnot last specially thanks to my family and friends. I am especially indebted to my parents for their love, sacrifice and support. They are my first teachers after I came to this world and have set great examples for me about how to live, study and work.
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