Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30General SciencesThe Use of Molecular Marker Methods in Plants: A Review
English0107Thoungamba AmomEnglish Potshangbam NongdamEnglishDifferent DNA markers have been utilized in the last few decades as important molecular tools in plants for genetic relation studies among individuals, hybrid and varietal identification, phylogenetic relationship among species, gene mapping and tracking quantitative trait loci. These markers can be broadly classified into hybridization and PCR based markers. Restriction fragment length polymorphism (RFLP) represents the hybridization based marker; while PCR dependent includes more reliable and advanced polymorphic markers like amplified fragment length polymorphism (AFLP), inter-simple sequence repeats (ISSR), single nucleotide polymorphism (SNP), sequence related amplified polymorphism (SRAP), start codon targeted (SCoT) and inter-primer binding site (iPBS) among others. Functional markers (FMs) have also been developed from functionally characterised sequence motifs which are superior to random markers due to their complete linkage to trait locus alleles. With the advent of next generation sequencing (NGS) technology, excellent opportunities are offered for generating ample structural and functional genomic information in many important crops. The novel approach of genotyping-by-sequencing (GBS) employs NGS protocols for discovering and genotyping SNPs in many important crop genomes and populations. The present review focuses on the description of varied molecular markers, their methodologies, strengths and limitations as well as applications in plant breeding and genetic research.
EnglishPolymerase chain reaction, Autoradiography, Functional markers, Genetic polymorphism, Next generation sequencingINTRODUCTION
Knowledge of genetic structure and level of variation within and between plant populations is important for effective utilization and conservation of plants. Factors determining level and structure of genetic variation within plant species include evolutionary history characteristics, population density, mating system and mechanism of gene flow [1]. Although morphological characters, cytological and ethnological parameters have been used traditionally to characterize levels and patterns of diversity, these traits alone represent only a small portion of plant genome and are influenced by environmental factors [2]. These limit their utility in describing the potentially complex genetic structures that may exist within and between taxa [3]. Advances in biochemistry and molecular biology during the past few decades have helped to overcome these constraints by providing breeders with many powerful molecular markers [4]. Various molecular approaches for detection of genetic diversity have been devised in many plants including important horticulture crops using DNA based markers. These markersare generally independent of environmental factors and are more numerous than phenotypic characters providing clearinformation of underlying variation in the genome of an organism. The present review aims to describe different marker systems which are employed in plant identification, crop improvement, genome analysis, phylogenetic and population diversity studies.
Hybridization based DNA marker
Restriction fragment length polymorphism (RFLP) is the only marker system representing hybridization based marker. This involves the use of restriction enzymes and hybridization of the target fragment by labelled probe. It is based on the generation of different size DNA fragments due to digestion by restriction enzymes. Genomes of individuals belonging to same species will differ in DNA fragment production after restriction digestion as a result of point mutation, insertion/deletion, translocation, inversion and duplication. RFLP steps involve the cutting of genomic DNA by restriction enzyme generating different sized DNA fragments. 6 base pair cutter enzymes are most often used for RFLP analysis as they are cheaper and readily available and alsogenerate product range (200 to 20,000 bp) that can be conveniently separated on agarose gels [5]. The separated DNA fragments are transferred to nitrocellulose membrane by Southern blot technique [6]. Fragments of interest are identified by hybridizing with complementary radioactive labelled probe and specific banding pattern is visualized after autoradiography. The result obtained from RFLP technique depends on both restriction enzymes and number of probes. RFLP exhibits high reproducibility, codominant inheritance, easy data transferability between laboratories and provides locus specific markers. Disadvantages of the technique are time consuming, requirement of high quality and quantity of DNA, expensive radioactive probes, involvement of tedious Southern blotting method and necessity of prior sequence information for developing radio labeled probe. The RFLP markers have been employed for genetic diversity and population genetic study in varied plants like Quercus phellos [7], Saccharum spp.[8] and Vigna radiata [9].
PCR based markers
Polymerase Chain Reaction (PCR) technique was invented by Kary Mullis in 1983 [10]. It involves the use of Taq DNA polymerase obtained from Thermus aquaticus for exponential amplification of very small amount of target DNA. The prior sequence information of flanking sites of the target DNA is required to design an appropriate primer for amplification of selected nucleotide segment. In addition to primers and DNA polymerase, supplementation of deoxynucleotide triphosphate (dNTPs) and magnesium ions along with appropriate buffer system are essential in basic PCR protocol. The PCR based markers are more advantageous over hybridization method due to less amount of DNA requirement, absence of radioisotopes, high reproducibility, more reliable and higher polymorphism in short time.
Amplified fragment length polymorphism (AFLP)
The technique employs both RFLP and PCR by ligating primer recognition sequences to the DNA fragments produced through restriction digestion [11]. Simultaneous screening of representative DNA regions distributed randomly throughout genome is the important feature of AFLP. Polymorphisms of AFLP may be produced due to mutation at restriction site, insertions, duplications or deletions inside amplification fragments and mutations of the sequences flanking the restriction sites. Good quality DNA as well as partially degraded DNA can be used for AFLP analysis but DNA should be free from restriction enzymes and PCR inhibitors. The genomic DNA might be digested by restriction enzymes which are the combination of rare cutter (Eco RI or Pstl) and frequent cutter (Msel or Taql). The double stranded oligonucleotide adaptors which do not bear initial restriction sites after ligation are developed and ligated to both ends of the fragments to give known sequence for PCR. PCR is first performed with primer combinations containing a single base pair extension while final amplification is carried out by using primer pairs with upto 3 base pair extension. AFLP fragments generated are visualized either on agarose gel or on denaturing polyacrylamide gel with autoradiography or AgNO3 staining respectively. The method is highly reproducible and its ability to determine high polymorphism in a single reaction makes AFLP one of the most sought after molecular tools for genetic analysis [12]. The limitations are the requirement of high molecular weight purified DNAs, possibility of co migrating non-homology fragments belonging to different loci.
Inter simple sequence repeat (ISSR)
This technique was first reported by Zietkiewicz at al. [13] and involved amplification of DNA segments present at an amplifiable distance between two identical microsatellite repeat regions oriented in opposite directions. Microsatellites of di, tri, tetra or penta-nucleotide core sequences are used as primers to amplify mainly inter simple sequence repeats of different sizes. ISSR primers are longer (15-35 mers) which allow higher annealing temperature resulting in higher stringency. The annealing temperature however depends on the GC content of the primer. After PCR amplification the amplified products of 200 to 2000bp long are separated through agarose or polyacrylamide gel electrophoresis and the resulting ISSR banding pattern can be visualized through autoradiography or AgNO3 staining. Prior sequence information of template DNA is not required for generating ISSR polymorphism. ISSR markers are simple, randomly distributed in the genome, exhibit mostly dominant inheritance pattern and require low quantity of DNA. They are employed in species and plant varietal identification, taxonomic and genetic diversity studies, gene mapping and clonal fidelity testing of in vitro derived plants[14]. The main limitations of ISSR marker are low reproducibility and homology of co-migrating amplification products [15].
Single nucleotide polymorphism (SNP)
It is the new generation molecular marker which detects polymorphisms among individuals due to changes in single nucleotide position. The simplest approach to discovering SNP in targeted region containing genes is performing direct sequencing of genomic PCR products derived from varied individuals. This approach may be costly for large scale study as there is a need for locus-specific primers and is limited to regions for which data are available. The other method based on the comparison of sequences obtained from cloned fragments can be considered for developing SNP map of the genome [16]. Many SNP genotyping methods are developed which combine two elements: generation of an allelic product and analysis thereof [16]. Sobrino et al. [17] assigned the majority of SNP genotyping assay to one of four groups based on molecular mechanism such as allele specific hybridization, primer extension, oligonucleotide ligation and invasive cleavage. Allele specific hybridization is based on distributing between two DNA targets differing at one nucleotide position by hybridization [18]. Two allele-specific probes are designed with polymorphic base in a central position in probe sequence. Under optimized assay conditions perfectly matched probe target is stable while one base mismatch is unstable. Most hybridization techniques are based on Dot Blot and Reverse Dot Blotmethods. Main advantages of the marker system are possibility of using small and extremely degraded DNA sample, high genomic abundance, completely automated sample processing and generation of no stutter products. The SNPs are found to be associated with plant genes of economic values e.g., presence of SNP markers for waxy gene controlling amylose content in Rice, linking of SNP with dwarfing gene in Rice and male sterility in Onion [19].
Diversity arrays technology (DArT)
This technique is based on microarray hybridization involving simultaneously genotyping of several hundred polymorphic loci spread over the genome [20]. Genomic representations are set up by genomic DNA restriction digestion followed by ligation of restriction fragments to adaptors for each of the individual DNA sample. Primers designed for adaptors and selective overhangs are used in PCR amplification in order to reduce genome complexity. DNA fragments derived from the representations are cloned and clone amplification is performed using vector-specific primersfollowed by purification and finally arrayed onto a solid support (microarray) forming a discovery array [15]. Labeled genomic representations formed from individual genomes included in the pool are hybridized to discovery array. Polymorphic clones exhibiting differential hybridization signal intensities are later set up into a"genotyping array" for routine genotyping[20].The advantages of this marker are high reproducibility, readily expandable, quick and non-requirement of prior sequence information. They are technically more demanding with long tedious steps and require skilled personnel being microarray-based technique. High investment in terms of time, physical energy, cost and advanced lab facilities limit the application of DArT markers in plant research.
Conserved DNA derived polymorphism (CDDP)
The technique was developed by Collard and Mackill [21] to fingerprint rice varieties. It was also used to evaluate the genetic diversity existed in Solanum dulcamara [22]. The short conserved sequences found in plant genome in multiple copies are targeted by primers designed to bind to these genes and generate polymorphic banding patterns. The primers also target common plant genes which are related to abiotic and biotic stress or responsible for plant development. Variation can be detected as length polymorphism within these regions since highly conserved DNA regions sharing the same priming site but differ in their genomic distribution. Single long primer amplifications with a high annealing temperature improve the reproducibility in CDDP. The DNA fragments produced are in the range of 200-1500 bp which are separated by electrophoresis and the banding patterns observed after autoradiography.
Start codon targeted (SCoT)
This is a novel marker system developed based on the short conserved regions flanking the ATG start codon in plant genome. SCoT involves the use of 18-mer primers with annealing temperature at 50oC [21]. Single primer is used in PCR which means the same primer is utilized as forward and reverses primer as in RAPD and ISSR markers. These markers are generally reproducible and the length and annealing temperature are not the most important factors determining reproducibility [23]. They are dominant markers which can be used for plant genetic analysis, quantitative trait loci (QTL) mapping and bulk segregate analysis [21]. The PCR products generated after amplification reaction are subjected to general agarose gel electrophoresis to separate the fragments and the bands are detected through autoradiography.
Sequence related amplified polymorphism (SRAP)
This simple, inexpensive, dominant marker technique was developed by Li and Quiros [24]. They developed the marker to specifically amplify coding regions of genome of Brassica oleraceae by targeting GC rich exons and AT rich promoters, introns and spacers. Forward primers are designed to contain GC rich sequences near the 3' end whereas reverse primers contain AT rich sequences at the 3' end."CCGG" sequences are present in the core of forward primers while the"AATT" sequences represent the core of reverse primers [25]. The PCR amplification products are resolved following agarose gel electrophoresis and the bands are visualized through autoradiography. The DNA fragments are scored by simple absence or presence of bands as done in ISSR and RAPD markers. SRAPs have been employed to evaluate genetic variation at species level, in population genetic analysis of closely related hybrids, construction of linkage maps and in identification of quantitative trait loci [26].
Inter-Primer binding site (iPBS)
Use of retrotransposons as molecular marker has limitation due to requirement of sequence information of LTR to design element specific primers. There is necessity for numerous cloning and sequencing steps to obtain a few good primer sequences. Most LTR regions do not have conserved motif for direct amplification by PCR [27]. Kalender et al. [28] developed iPBS marker for identifying diverse LTR sequences and directly visualising the polymorphism among the plant cultivars. This method utilizes PBS (primer binding sites) which is a conserved sequence located adjacent to the 5'LTR and is universally present in all retrotransposons. The tRNA binds to PBS region to initiate reverse transcription by producing complementary base pairing between terminal sequence of tRNA and conserved region of PBS [29]. Primers can be designed which match the conserved region of PBS and DNA fragments of diverse LTR sequences can be generated after PCR amplification [27]. The cloning of LTR sequences was previously dependent on conserved protein coding domains limiting the screening of autonomous elements [30]. But iPBS technology allows screening of diverse LTR sequences while performing effective DNA fingerprinting. The level of polymorphism generated by iPBS amplification is as efficient as that of IRAP and RBIP markers. TheiPBS markers are highly effective and reliable for polymorphism detection and determination of clonal differentiation arising out of varied retrotransposon activities and retrotransposon recombinations [28]. iPBS is also highly reliable and powerful DNA finger printing technology which does not require the knowledge of sequence information [31]. It is used in phylogenetic and genetic diversity studies of number of plant species. Genetic diversity studies have also been successfully performed using iPBS markers in Apricot, Lens species and field pea [31, 32]
FUNCTIONAL MARKERS (FMs)
Morphological markers previously employed in plant breeding are easily affected by the environment apart from their limited number and difficulty in scoring [33]. The introduction of random DNA markers RFLP, SSR,AFLP and SNPs have brought significant improvement in plant breeding programs for producing improved crop varieties. They are used for hybridity confirmation, parents and progenies identification, evolutionary relationship study, genetic diversity determination between and within populations and mapping genes for marker assisted selection (MAS). However, the use of such markers as diagnostic tool for MAS in plant breeding has limitations because of false positives produced by genetic recombinations[34]. Genetic recombination between the marker and target locus impairs the transfer of marker information from experimental mapping population to unrelated breeding materials [35]. The random markers are derived at random from polymorphic sites in the genome and are developed independent of their relationship to any phenotypic characters. But functional markers (FMs) are developed from polymorphic sites within the genes casually involved in producing phenotypic trait variation [33]. The absence of recombination for FM and its complete linkage to desired allele prevent information loss and false selection in MAB [36]. This also makes FM more effective in identification and selection of favourable alleles and enhances its diagnostic capability. Unlike random DNA markers, the phenotypic validation in MAB is not required while using FMs in plant breeding [33]. When random markers are used in MAB, there is possibility of transferring target gene along with unwanted genes located at a distance due to linkage drag producing undesirable phenotypic traits[37].
Functional marker development
The development of functional markers requires functionally characterised genes and identification of phenotypic/functional sites affecting plant phenotypic traits. One of the challenges of functional marker development is to relate sequence polymorphic of functional motif with phenotypic trait variation [33]. Association studies identify genes and even functional motif within the genes that affect phenotypic characters [38]. But this approach is dependent on linkage disequilibrium (LD) mapping which rely on non-random occurrence of allele haplotype in the genome[39]. Low level of LD is essential for determining the effects of intragenic polymorphism on phenotypic changes. Association studies can identify sequence motifs affecting phenotypic trait expression in crops having low LD. Application of association studies may not suffice the distinction of causative from phenotypically neutral polymorphism in haplotype structure [38]. Functional markers developed through the involvement of association studies are termed as indirect functional markers (IFM) as only indirect (statistical) evidence of sequence motif function can be provided [33]. However, the direct proof of sequence motif function can be achieved after comparing the isogenic genotypes which differ in single sequence motif. These isogenic genotypes can be generated through the approach of TILLING and homologous recombinations [33]. The development of FMs for selected traits has been observed successfully in few crops. Iyer and McCouch [40] cloned gene xa-5 in rice which made it possible to develop functional markers for Xa5-mediated resistance to bacterial blight disease [41]. Several candidate genes for FM development which have potential for controlling agronomic traits have been cloned in different plant crops - Dwarf 8 in Maize [38] and OsARD2 in Rice [42].
GENOTYPING-BY-SEQUENCING (GBS)
The traditional DNA sequencing technologies could not meet the demand for indebt sequence information required in complex genomic research. With the advent of next generation sequencing technology (NGS), the knowledge gap is filled and the technology has become an inevitable everyday research tool in complicated genome studies [43]. Before the NGS came into being, Sanger or dideoxy sequencing was the most widely used DNA sequencing technique. But the Sanger sequencing is relatively expensive, laborious and time consuming apart from its inherent limitation of requiring in vivo amplification of DNA fragments to be sequenced. The limitations are overcome with the introduction of 454 technology in the market which was the first NGS technology which relied on the method of in vitro DNA amplification known as emulsion PCR [44]. 454 platform marketed by Roche Applied Science has the capability of generating 80-120 Mb of sequence in 200-300 bp reads in 4 hour runs and is one of the most popularly used sequencing technologies in genome research. Recent advancements in NSG technologies make it possible the use of SNPs for genetic analysis to a new level. Genotyping-by-sequencing (GBS) first introduced by Elshire et al. [45] is a novel application of NSG protocols for discovering and genotyping SNPs in crop genome and populations. This is highly multiplexed system for constructing reduced representation libraries for the Illumina NGS platform generating good numbers of SNPs for employing in genetic analysis and genotyping [46]. GBS is a cost effective and efficient technique for genomics assisted breeding in varied crops like Cotton, Brassica, Sorghum and Miscanthus [43]. Using ion PGM systems, two GBS strategies have been developed [47]. The first approach is ideal for discovering new markers for MAS programs and no specific SNPs are identified with digestion by restriction enzymes. Genome complexity is reduced with DNA restriction digestion using one or two selected restriction enzymes before adapter ligation [48]. In the second approach, a set of SNPs is defined for particular genome region using PCR primer designed to amplify only the selected interest region of genome [47]. The use of sequencing restriction site associated genomic DNA (RAD) for discovery of high density SNP and genotyping is more complex and expensive as compared to GBS.
CONCLUSION
The varied molecular markers employed in plant research have been discussed along with their applications in genetic variability evaluation, genome fingerprintingand population genetic studies. Hybridization based DNA marker which is one of the pioneer markers for plant genetic diversity studies have been superseded by the development of more reliable and reproducible PCR based markers. However, either simple or more advanced molecular markers possess inherent strengths and weakness and none of them are perfect with shortcomings. Selection of the most appropriate marker will ultimately depend on the particular research approach adopted and degree of resolution and polymorphism required for the specific study. With rapid progress in molecular biology, more effective and superior markers may appear in near future which can significantly accelerate plant breeding research. The recent advancement of NGS technology which led to the development of GBS will also provide ultimate MAS tool for rapid enhancement in plant improvement and crop breeding.
ACKNOWLEDGEMENT
The authors are thankful to SERB (Science and Engineering Research Board), New Delhi, India for the financial support. Authors also 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 the articles, journals and books from where the literature for this article has been reviewed and discussed.
CONFLICT OF INTEREST
The authors declare that there is no conflict of interest regarding the publication of the paper.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30General SciencesForest Invasive Species Assessment Study in Different Village Forests of Garhwal Himalaya
English0818Arti KhanduriEnglish Sas BiswasEnglish H.B. VasisthaEnglishForest invasive species (FIS) are exotic/alien species that occur outside their natural adapted ranges and dispersal potential. Some of the alien species become invasive, when they are introduced deliberately or unintentionally outside their natural habitats into new areas, where they express the capability to establish, invade and outcompete native species. The present study is focuses over the encroachment of invasive species in the two different forest communities of the Tehri Garhwal region of Western Himalaya. Data was collected through extensive field survey and quadrat method. High invasion was recorded in the shrub and herb layer of the forest. In tree strata native species are dominant but their recruitment in the form of sapling and seedlings are displaced by the dense thickets of invasives in both the communities. A highest value of ecological indices was evaluated in Pinus roxburghii dominated site as compared to the Quercus leucotrichophora dominated site. Lantana camara, Eupatorium glandulosum, Clematis gouriana, Rosa brunonii, Rubus neivus, Euphorbia royleana etc. are the most destructive Forest Invasive Species (FIS*) of both the forest communities. The present study gives an accurate assessment and understanding of the dynamics of invasives, which is further important for their scientific management and utilization.
EnglishWestern Himalaya, Biological invasion, Invasive species, Plant community, DominanceIntroduction
India is one of the 17 mega biodiversity countries of the world. Diverse geographical and edaphic conditions of the country provide the luxuriant growth for various kind of flora and fauna (Chaudhry, et.al., 2011). This diverse ecological condition provides platform for different new (alien) species to come up. Being rich in ecological diversity, forest ecosystem of the Himalayan region gives high value of ecosystem services. Its diversified landforms, relief and climatic zones support a wide range of vegetations. (Rana et.al., 2010). In last century, the ecology of the Himalayan region has changed to a considerable level. Increase in the overall temperature of earth atmosphere and especially Himalayan region has changed the rainfall as well as seasons. Damming of rivers in Himalayan region has changed the ecology of the adjacent areas to lacusterine from riverine, resulting in change in flora of the region (Adhikari et. al., 2009). This climatic as well as ecological change is also responsible for encroachment of the region by new (alien) species. According to the Convention on Biological Diversity, invasive alien species are the second largest cause of biodiversity loss in the world and impose high costs to agriculture, forestry, and aquatic ecosystems (Raizada et.al.,2008). The global extent and rapid increase in invasive species is homogenising the world's flora and fauna (Mooney and Hobbs, 2000) and is recognized as a primary cause of global biodiversity loss. Bio-invasion may be considered as a form of biological pollution and significant component on global change and one of the major causes of species extinction (Mooney and Drake, 1987).
Persistent colonies composed of invasive woody perennials alter the structure and function of forest ecosystems by inhibiting the growth and development of native species (Webster et al., 2006). The ability of these species to inhibit the recruitment of native species raise the possibility that large-scale invasions may fundamentally change the successional trajectory of forest ecosystems. Such a change would have far-reaching implications for numerous plant and animal species that rely on native plant communities and their successional pathway (Webster et. al., 2006).
Forest invasive species are thus a serious hindrance to conservation and sustainable use of biodiversity significant undesirable impacts on the goods and services provided by ecosystems. It is reported that losses due to alien invasive species in the country amounts to about US$130 billion annually.Forest alien species is now at global scale and it is expected that it will undergo rapid increase due to interaction with other changes such as globalization of markets, rise in global trade, travel and tourism (McNeely, et.al.,2001).
Our lack of knowledge about how invasive species function in their new environment significantly undermines our ability to detect and eradicate new or small infestations, therefore the effective management of invasive species, is only possible by gathering knowledge about their ecology, morphology, phenology, reproductive biology, physiology and phytochemisty , so that we can find and eliminate new infestations.
Material and Methods
Study area
The State of Uttarakhand lies between 28044’ to 31028’ N latitude and 77035’ to 810 05’ E longitude, encompasses an area of 53,483 Km2. The Garhwal Himalaya encompasses all 4 of the Himalayan physiographic-geological zones. These are broadly classified into Shiwalik, Lesser Himalaya, great Himalaya and Trans Himalaya.
The study was carried out in the District Tehri Garhwal of Uttarakhand, which lies between 3001’ to 3009’ N latitude and 7709’ to 7901’ E longitude in Garhwal Himalaya and covers an area of 3796 sq. km of area. The region is highly mountainous, ranging from 300-7000m in altitude. The sites selected for the study were Jardhar village forest ecosystem and Kuttha village forest ecosystem. Both the areas selected for study are part of the lesser Himalayan ranges. Both are situated on north-eastern aspect, thus, received almost same amount of insolation. The two study sites have different altitude level and ecology. The Jardhar Village study area lies between the altitudinal range of 1400 mtr to 1800 mtr, whereas,the Kutha Village study site lies between 1000 to 1400 mtr altitude. Apart from altitudinal variation in sites, there is one more important feature of Kutha Village site i.e, it is part of the peripheral slope of Tehri Dam Reservior.
Phytosociological assessment
To assess the extent of invasives in the forest area, a vegetation/phytosociological study was carried out. The vegetational surveys were conducted in the pre-monsoon and post monsoon periods of the study year. Detailed field surveys were adopted for recording the floristic composition of the communities in these sites.
The vegetation sampling was carried out, in order to reveal the quantitative impact in terms of IVI value, occurrence and area of occupancy of FIS in different sites.
Quadrat method was used to sample the vegetation. Size of quadrats used during the study were, 10m x10m, 5m x 5m, and 1m x 1m (Mishra, 1968), to enumerate trees, shrubs and herbaceous plants respectively. In Site I (Jardhar village forest) 120 quadrats were laid whereas in Site II (Kuttha village forest) 50 quadrats were found sufficient.
Individuals between 10.5 to 31.5cm cbh (circumference at breast height i.e., 1.37m above ground level) were recorded as shrubs and individuals less than 10.5cm cbh were considered as herbaceous plant (Knight, 1963).
The seedlings were considered as herbs and saplings as shrubs. Elevation/altitude, longitude, latitude of the site was recorded by GPS at 5-6 accuracy.
The Phytosociological data were quantitatively analysed for frequency, density and abundance following Curtis and McIntos (1950) and Mishra (1968). Importance Value Index (IVI) was determined as the sum of the relative values of frequency, density and dominance. The relative values of these parameters Relative frequency (RF), Relative density (RD) and Relative dominance (RDom)) were determined following Phillips (1959). Abundance (A)/ frequency (F) ratio, (Whitford, 1949) was used to interpret the distribution of the species. The distribution was classified viz.,-regular (0.05) following the method of Curtis and Cottam (1956).
FREQUENCY, DENSITY AND ABUNDANCE
Frequency refers to the degree of dispersion of individual species in an area and is usually expressed in terms of percentage occurrence. It can be calculated as:
Frequency % (F %) = No. of quadrats of occurrence of a species x100
Total no. of quadrats studied
Density (D) = Total no. of individuals of a species in all the quadrats
Total no. of quadrats studied
Abundance (A) = Total no. of individuals of a species in all the quadrats
Total no. of quadrats in which species occurred
Relative frequency (RF) = No. of occurrence of the species x100
No. of occurrence of all species
Relative density (RD) = No. of individuals of the species x 100
No. of individuals of all species
Basal Area: This is regarded as an Index of dominance of a species. Higher the basal area, greater is the dominance and this is calculated by the term of relative dominance.
Rel. Dom (RDom) = Total basal area of the species in all quadrats x 100
Total basal area of all species in all the quadrats
A total picture of the ecological status of a species with respect to a particular community structure can be obtained only after calculating the values of RF, RD, RDom. These values when added together given the importance value index (IVI).
IVI= Rel.frequency +Rel.density +Rel.Dominance
Results
The most important constituents of vegetation are plant species and their assemblages, which depend on various factors such as altitude, aspect, soil, geology, topography, natural herbivores and anthropogenic activities (Mueller-Dombois and Ellenberg, 1974). The functioning of an ecosystem is determined by the components of a community. They vary in quality and quantity in a given time and space, and has impact on the structure of the community. Therefore structure and composition of the vegetation reflect the ecosystem properties and ecological conditions of an area and form the basis for further scientific and management studies (Lindenmayer and Franklin, 1997). This paper deals with the ecological status of the sites in terms of plant structure, composition and invasion level along the native species in the affected village forests.
ECOLOGICAL STATUS OF SITE I (JARDHAR VILLAGE FOREST ECOSYSTEM)
Tree layer: A total of fifty six tree species were recorded in the site. Out of these, 55 species were broad leaf and one species, i.e., Pinus roxburghii was conifer. The most dominant tree species of the forest was Quercus leucotrichophora followed by Rhododendron arboreum and Pinus roxburghii .While, in terms of invasion only one tree species Euphorbia royleana was reported as invasive. The species was distributed randomly along with native species of the area. (Fig 2 and Table 1)
Forest Invasive Species*
Shrub layer: A total number of 72 species were recorded under shrub layer. Out of them 39 species were shrubs and 33 species of trees in the form of saplings. The most dominant species under this category was Lantana camara followed by Rubus ellipticus and Myrisine africana (Fig 3, Table 2).
Fig.3. Dominant shrub species of site I (Jardhar VFE)
Invasion status of shrub layer:
Shrub layer was encroached by total a no. of 11 invasive species. The most dominating invasive species were Lantana camara followed by Rubus ellipticus and Rosa brunonii.( Fig 3 and Table 2).
Forest Invasive Species*
Herb layer: Under herbaceous layer two life forms of plants, i.e., herbs including invasive herbs and seedlings were recorded. Total 67 species were recorded in this layer. Out of these, 47 species were herbs and 20 species were in the form of seedlings of trees and shrubs (Table 3).
The most dominant species was Eupatorium glandulosum followed by Oplismenus, seedling of Myrsine africana and Apluda mutica.
Invasion status of herb layer: The most dominating invasive was Eupatorium glandulosum followed by Apluda mutica, Anaphalis busua and Echinochloa colone. A/F values reveal that invasives follows random as well as contiguous pattern. (Fig 4 and Table 3).
Forest Invasive Species*
ECOLOGICAL STATUS OF SITE II (KUTTHA VILLAGE FOREST ECOSYSTEM)
Tree layer: A total no of 36 tree species including a single invasive species Euphorbia royleana were recorded from tree stratum of the forest in different composition.
The most dominant species was Pinus roxburghii 2nd most dominating was Rhus parviflora followed by Quercus leucotrichophora, while Euphorbia royleana recorded as the 4th most dominating species (Fig 5 and Table 4).
In terms of invasion level, only single species Euphorbia royleana was found to be among tree layer. The species are contiguously distributed in the forest as shown by A/F values.
Shrub layer: The total species under shrub layer was recorded 39 in number (Table 5). Out of them 21 species were shrubs including invasives and 18 were tree species in the form of saplings. The most dominant species among shrubs was Lantana camara followed by sapling of Rhus parviflora, Carissa opaca and sapling Euphorbia royleana.
Invasion status of shrub: Shrub layer of Kuttha village forest ecosystem was invaded by 8 shrubby invasive species in different phytosociological attributes. Level of invasion was observed maximum for Lantana camara followed by saplings of Euphorbia royleana, and Rubus ellipticus, while the least dominating species was Clematis gouriana (Fig 6 and Table 5).
Herb layer: In the site study a total no. of 51 species were recorded under herb layer (Table 6) among them 11 no. of species were counted as seedlings of trees and shrubs whereas 40 were of herbaceous species including invasives. The most dominating herb was Eupatorium glandulosum co-dominated by Parthenium hysterophorus followed by seedling of native species Rhus parviflora.
Invasion status of herb layer: A total no. of 34 herbaceous invasive species was recorded from the forest (Table 6). The most noxious species was Eupatorium glandulosum followed by Parthenium hysterophorus and Ageratum conyzoides in herb layer.
These species are distributed in contiguous pattern in the herb layer.
Discussion
Many plant species serve as indicators of ecological conditions and have been used for site evaluation (Rowe, 1956).
The present study was conducted in two different sites. i.e. Jardhar villege forest and Kuttha village forest. The study reveals that both the sites are invaded by Forest Invasive Species in all the layers of the forest ecosystem. The level of invasion in terms of their dominance, density and basal cover varies as per site conditions. As in Jardhar village forest, Quercus leucotrichophora, Rhododendron arboreum, Pinus roxburghii etc. were the dominant trees, whereas, the Kuttha village forest is occupied by Pinus roxburghii, Rhus parviflora, Quercus leucotrichophora etc, also invaded by Euphorbia royleana. It indicates that both the forest types are infested by invasives but the level of infestation is high in Pinus roxburghii dominated site. Pinus roxburghii is the pioneer species in low soil depth and nutrients deficient degraded slopes These soil conditions provokes the growth of Euphorbia royleana, as also suggested by other workers (Munesh K et.al., 2017, Adhikari et.al., 2009, Banerjee, 1986). Euphorbia royleana infestation was also reported from Quercus leucotrichophora dominated site but its expansion was reported from the eroded and degraded rocky pockets of the forest. But was less as compare to the Pinus roxburghii dominated sites, as shown in the study Table 1 and 4.
In shrub layer, the dominance of Lantana camara, a shrubby invasive in both the sites has resulted in the replacement of native species of the region (Love et. al., 2009). Thus native species are now being treated as secondary species. Spreading foliage of Lantana camara, which are making impenetrable thickets, covers the ground fully and promoting shade condition, which prepare the ground for the shade or moisture loving species. The tendency of this invasive though sometimes favours the shade and moist loving native species but such areas are found to be encroached by shade loving invasive Eupatorium glandulosum, as its dominance in herbaceous layer suggest this fact; therefore it is presumed that dominance of Lantana camara in shrub layer indirectly making the encroached area more prone for further invasion (Lau 2008, Holzmueller and Jose, 2009, Usher 1991), and the infestation in phytosociological terms was recorded high in Pinus roxburghii dominated site as the site is more prone (Uniyal 1995) .
Researcher observed that some plants such as Lantana camara and Euphorbia royleana altogether occupying and multiplying rapidly in dry, eroded and disturbed localities (especially Kuttha village forest ecosystem). These species are reducing the native vegetation of these slopes as the same have been reported by (Banerjee, 1989). Similarly Eupatorium glandulosum, Parthenium hysterophorus and Ageratum conyzoides are quickly moving towards the inner ranges of the forest as revealed from phytosociological analysis of area and the same have also been reported from district Uttarkashi and Pauri Garhwal by Gaur (1999), thus presumed that the natives of these encroached elevation certainly get displace due to current exisistence and dominance of forest invasive species.
Silvicultural practices (Troup, 1921) reveals that the nature of the maximum natives (trees, shrubs and herbs, except Pinus roxburghii) and invasive Eupatorium glandulosum, Rosa brunonii, Clematis gouriana, etc. are moisture loving whereas invasive Lantana camara, Euphorbia royleana grow well in all kind of conditions (Howard, 1969; Banerjee, 1989), thus all moist pockets of the forest where native species could recruit (especially as in Jardhar village forest) are encroached by these invasives, and by virtue of specific traits and characteristic features (Reddy et. al., 2002; Love et al., 2009) these species became good competitor over native species, therefore natives though recruit but least in dominance as found to be replaced by invasive shrubs and herbs hence creating loss of native biodiversity as also studied by Sharma et al., 2005a; Day et al., 2003; Webster et al., 2006). (Table 1 to 6 and Fig. 2 to 7).
CONCLUSION: The study indicates that site, which is Pinus roxburghii dominated, is under threat and more prone to further invasion as compared to the Quercus leucotrichophora dominated site. High level of invasion in the shrub and herb layer imply towards the immerging threat to native biodiversity of the study area. Therefore, there is an urgent need to check, monitor, and evaluate the further invasion and needs to apply the scientific corrective measures.
ACKNOWLEDGMENT: Authors are grateful to the Forest Research Institute, Dehradun. We 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.
Englishhttp://ijcrr.com/abstract.php?article_id=2322http://ijcrr.com/article_html.php?did=2322Adhikari BS, Uniyal SK, Rawat GS. Vegetation structure and community patterns of Tehri Dam Submergence Zone, Uttarakhand, India. EurAsia J BioSci 2009; 3, 40-49.
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ay M, Wiley CJ, Playford J, Zalucki MP. Lantana: current management status and future prospects. ACIAR Monograph, 2003; 102, 28.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30General SciencesEffect of Integrated Nutrient Management on Essential Oil (Volatile Oil) of Coriander (Coriandrum sativum L.)
English1925Sunita JhariyaEnglish Aruna JainEnglishAim: The experiment was intended to find out the effect of Integrated Nutrient Management on Essential oil (Volatile oil) of Coriander (Coriandrum sativum L.).
Methodology: The experiment was laid out in a randomized block design with 8 treatments using chemical fertilizers (NPK), vermicompost and biofertilizers (Azotobacter and Phosphate Solubilizing Bacteria sp. Pseudomonas strata) in different combinations including one control treatment.
Results: The results indicated that volatile oil percentage and percentage of different compounds i.e. Linalool, Geraniol, a-pinene and B - pinene in volatile oil of Coriandrum sativum L. was recorded significantly higher in T7 treatment compared to control treatment at different growth stages.
Conclusion: From the analysis of results it can be concluded that integrated use of biofertilizers, chemical fertilizers and vermicompost is the best combination for the Coriander (Coriandrum sativum L.) crop.
EnglishBiofertilizers, Chemical Fertilizers, Vermicompost, Integrated Nutrient Management, Volatile oil, Coriandrum sativum
Introduction
India is known all over the world as"The Home of Spices". The fame of Indian spices is older than the recorded history. The term spice applies to natural plant or vegetable products or mixtures in whole or ground form, which are used for imparting flavor, aroma and piquancy to the food items. Spices are made up of many parts of a plant such as roots, bark, flowers, fruits, leaves etc. with distinct flavor and taste (www.mccormickscienceinstitute.com). Coriander (Coriandrum sativum L.) an annual herb of the parsley family (Apiaceae), is native to the Mediterranean region and is extensively grown in Bangladesh, India, Russia, central Europe and Morocco and has been cultivated since human antiquity (Small,1997 and Bhuiyan et al., 2009). The plant is grown widely all over the world for seed, as a spice, or for essential oil production (Lawrence, 1993). All part of the plants is edible but the fresh leaves and the dried seeds are the most common parts used in cooking. Coriander contains an essential oil (0.03-2.6%). The essential oil of coriander is also called volatile oil. It has strong effects on the nervous system and is therefore widely used by aroma therapists and herbologists as a sedative, spasmolytic and local anaesthetic. It is also used against many skin complaints, mostly in the form of tea tree oil. The different parts of this plant contain monoterpenes, limpnene, a-pinene, g-terpinene, p-cymene, citronellol, borneol, camphor, coriandrin, geraniol, dihydrocoriandrin, coriandrons A-E, flavonoids and essential oils (Farooq Anwar et al., 2011 and Nadeem et al, 2013). Various parts of this plant such as seed, leaves, flower and fruit, possess antioxidant activity, anti-diabetic activity, anti-mutagenic activity, anthelmintic activity, sedative - hypnotic activity, anticonvulsant activity, diuretic activity, cholesterol lowering activity, protective role against lead toxicity, antifungal activity, anti-feeding activity, anticancer activity, anxiolytic activity, hepatoprotective activity, anti-protozoal activity, anti-ulcer activity, post-coital anti-fertility activity, heavy metal detoxification (Momin et al., 2012). According to Polo and De Bravo (2006) Geraniol has anti-tumor activity against human hepatocarcinoma cell line Hep G2, human pancreatic cancer cells (Jin et al., 2013) and anti-proliferative effects on human colon cancer cells (Carnesecchi et al., 2002). Chen et al., (2015) found that liver cancer cell growth was inhibited by the use of α - pinene. Silva et al., (2012) found that α and β - pinene has antimicrobial activities against bacterial and fungal cells. All parts of the plant contain essential oils that give the green plant a characteristic smell, which is similar to that of bugs. This smell is caused by different aldehydic components of the essential oil present in the green plant.
Material and Methods
The field experiment was conducted in farmer's field at village Raghogarh, Distt. Guna, Madhya Pradesh. The experiment was conducted in a randomized block design (RDB) with 8 treatments and three replicas of each, using vermicompost, chemical fertilizers (NPK) and biofertilizers (Azotobacter and Phosphate Solubilizing Bacteria) in different combinations including one control treatment. The treatments were T1 - Biofertilizers (250g Azotobacter + 250g PSB ha-1), T2 - Vermicompost 5t ha-1, T3 -Chemical Fertilizers (60:30:30 kg NPK ha-1), T4 - Biofertilizers + Vermicompost ( 125g Azotobacter + 125g PSB + 5t Vermicompost ha-1 ), T5 - Vermicompost + Chemical Fertilizers [5t Vermicompost ha-1 + 50% NPK ha-1 (RDF)], T6 - Chemical Fertilizers + Biofertilizers [50% NPK ( RDF as 30:15:15 kg per ha-1) + 125g Azotobacter + 125g PSB ha-1], T7 - Biofertilizers + Vermicompost + Chemical Fertilizers [125g Azotobacter + 125g PSB ha-1 + 5t Vermicompost ha-1 + 50% NPK (RDF as 30:15:15 kg ha-1)], T8 -Control (No Treatment).
Estimation of Essential oil (volatile oil)
Estimation of essential oil of Coriandrum sativum was performed on vegetative parts and on harvested dried seeds. The extraction of volatile oil was done by hydrodistilation for 4 hour, using Clevenger's apparatus to know the percentage of volatile oil and analysis of volatile oil was done by using GC -MS (Gas Chromatography - Mass spectrometry) to know the composition and to know the quantity of each component in volatile oil as described by Masada, 1976.
The oil dried over anhydrous sulphate solution, and stored at 4°C up to analysis by GC-MS.
Analysis of essential oil
Analysis of essential oil was done by using Gas Chromatography with Mass Spectrometer as described by James and Martin (1951) to know the composition of oil and the quantity of each composition. Analysis of essential oil for chemical composition was carried out using an Agilent -Technologies 6890 N network gas Chromatographic (GC) system equipped with 5975 inert XL mass selective detector and 7683 B series auto injector Agilent -Technologies. A sample volume of 1.0 µl was injected, applying split mode (split ratio 100:1), into HP-5 MS capillary column (30m x 0.25 mm), film thickness 0.25 m.
The initial column temperature was set at 800C and raised to 2200C by the rate of 40C/ min. The initial and final column temperatures were held for 3 and 10 min, respectively. The operating temperatures for detector and injector were 2200C and 2900C, respectively. The mobile phase used was helium at a flow rate of 1.5 ml/min. An electron ionization (EI) system, with ionization energy (70 eV) was used for GC-MS detection. Mass scanning range was varied over 50 to 550 m/z. the injector and MS transfer line temperature were 2200C and 2900C, respectively. The essential oil compounds were identified on the basis of matching their retention indices in relation to n-alkenes (C9 -C24) and moreover with those of authentic compounds or published data (Minica et al., 2004, Vagionas et al., 2007). Beside the comparison of MS spectral data of the compounds with those from NIST mass spectral library was also applied to authenticate the compounds (Adams, 2001).
Results
Various nutrient schedules influenced the essential oil % and % of different compounds in essential oil (volatile oil) of coriander at different growth stages. (Table and Fig. 1, 2, 3)
The combined application of BF 250g/ha + VC @ 5t/ha + 50% NPK RDF as 30:15:15 kg/ha (T7) was found to be superior when compared to control, in registering the maximum essential oil (%) in vegetative parts of the plant at different growth stages, while in dried seeds after harvest, it was also superior over control, when compared to other treatments it was found non-significant.
The amount of different compounds (α-pinene, β-pinene, Linalool and Geraniol) within the essential oil were also recorded higher in combined application of BF @ 250g + VC @ 5t/ha + 50% NPK RDF as 30:15:15 kg/ha (T7) when compared to control (T8) at different growth intervals.
Discussion
The essential oil % and % of different compounds in essential oil (volatile oil) of Coriander was found maximum in T7 treatment with the application of BF + VC + CF. The better essential oil (%) in plants might be due to increased vegetative growth due to integrated nutrient management that results in better photosynthetic activity and it results in increased volatile oil (%) in Coriander plants at different growth intervals.
The study by Kumar T. Senthil et al. (2009) also revealed that the combined application of nitrogen and phosphorus along with biofertilizers increased the essential oil yield in
Devana (Artimisia pallens Wall). The results are in conformity with Srinivasan et al. (2005) in black pepper, Garg (2007) in fennel (Foeniculum vulgare Mill.), Gharib et al. (2008) in Sweet Marjoram (Majorana hortensis).
Conclusion
The findings revealed that the application of BF 250g/ha + VC @ 5t/ha + 50% NPK RDF was effective and it significantly improved the essential (volatile) oil (%) and percent of different compounds in essential oil of Coriander. Therefore, the nutrient needs can best be met through integrated nutrient management (INM). The concept of INM aims to increase the efficiency of use of all nutrient sources, the soil resources, mineral fertilizers, organic manures and recyclable wastes or biofertilizers. It can be concluded by these findings that biofertilizers and vermicompost along with chemical fertilizers (RDF) should be applied to Coriander for better quality crop, as already reported by Darzi and Hadi (2012) in coriander (Coriandrum sativum L.).
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.
Englishhttp://ijcrr.com/abstract.php?article_id=2323http://ijcrr.com/article_html.php?did=2323
Adams, R. P. (2001). Identification of essential oil components by gas chromatography/quadrupole mass spectroscopy. Carol Stream IL, USA: Allured.
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Carnesecchi S, A Bradaia, B Fischer, D Coelho, M Schöller-Guinard, F Gosse and F Raul (2002). Perturbation by geraniol of cell membrane permeability and signal transduction pathways in human colon cancer cells. J Pharmacol Exp Ther, 303 (2): 711 -5.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30HealthcareThe Configuration and Variations in the Developing Circle of Willis- A Retrospective Autopsy Study
English2631Kanchan KapoorEnglish Jyotsna SinghEnglish Joseph AbrahamEnglish Amrutha Kavil ValapilEnglishIntroduction: Cerebral hemodynamics undergoes a shift from carotid system to vertebro-basilar system from fetal stage to adult life. The circle of Willis (CW) plays a major role in redistributing the blood and enables interhemispheric flow through the anterior and posterior communicating arteries (AComA, PComA). The text book description mention three stages in the development of CW- fetal, transitional and adult. The purpose of the present study was to observe development of arterial anastomoses during intrauterine life, to note the variations and trying to explain some of the hypothesis for these variations.
Material and Method: The material of the present study include 45 fetal brains of different age groups. Variations were observed regarding absence, duplication, bilateral asymmetry and tortuousity.
Results: The number and type of variations in fetus were lesser as compared to adults. Anomalies including presence of aneurysm, absence of anterior cerebral artery (ACA) and splitting of a constituent vessel were not observed in fetal brains. However there were incidences of absence / duplication and hypoplasia of a component vessel. During early gestation period, all the cerebral vessels were slender but uniform in size. This is named as fetal stage. After 22nd wks of fertilization, due to rapid development of occipital lobes, there is increase in the diameter of either posterior cerebral artery (PCA) or PComA thereby giving rise to either adult or transitional type of configuration respectively.
EnglishFetal brains, Circle of Willis, VariationsINTRODUCTION
The circle of Willis (CW) is formed at the termination of two internal carotid arteries and joining of vertebrobasilar trunk with its branches. All arteries of the forebrain arise from this anastomotic polygon. The circle of Willis (CW) plays a major role in redistributing the blood and enables inter-hemispheric flow through the anterior and posterior communicating arteries. The normal configuration of circle of Willis (CW) appears in a 52nd day embryo and its variations are well described in literature1. Although many cerebral arterial variants may be asymptomatic, their recognition is important in patients with cerebrovascular disease and in those who undergo brain imaging for other purposes, as some of these variants might be pathological or turn pathological in a surgical setting. Fenestrations and/or duplication of intracranial arteries have been associated with the development of brain saccular aneurysm 2.The present study is an attempt to observe the development of arterial anastomosis during intrauterine life and to note the variations. An effort has been made to explain the hypothesis for the occurrence of these variations.
MATERIAL AND METHODS
The material for the present study included 45 fetal brains in 14-30 weeks of gestation. The fetal brains were carefully removed and examined with an dissecting microscope during routine autopsy performed at Department of Anatomy, Government Medical College and Hospital, Chandigarh,
The circle of Willis (CW) is formed at the termination of two internal carotid arteries and joining of vertebrobasilar trunk with its branches. All arteries of the forebrain arise from this anastomotic polygon. The circle of Willis (CW) plays a major role in redistributing the blood and enables inter-hemispheric flow through the anterior and posterior communicating arteries. The normal configuration of circle of Willis (CW) appears in a 52nd day embryo and its variations are well described in literature1. Although many cerebral arterial variants may be asymptomatic, their recognition is important in patients with cerebrovascular disease and in those who undergo brain imaging for other purposes, as some of these variants might be pathological or turn pathological in a surgical setting. Fenestrations and/or duplication of intracranial arteries have been associated with the development of brain saccular aneurysm 2.The present study is an attempt to observe the development of arterial anastomosis during intrauterine life and to note the variations. An effort has been made to explain the hypothesis for the occurrence of these variations.
Material and methods
The material for the present study included 45 fetal brains in 14-30 weeks of gestation. The fetal brains were carefully removed and examined with an dissecting microscope during routine autopsy performed at Department of Anatomy, Government Medical College and Hospital, Chandigarh, India. The approval from the ethical committee of the institute was taken prior to the autopsy. The fetuses which were sent for autopsy were obtained from spontaneous or clinical abortions in the Obstetrics and Gynaecology department of the institute. Prior to the autopsy consent was taken from the parents. Cases with gross pathological lesions of the brain were excluded. After removal of brains, the cerebral vessels were examined at the base of the brain. Variations were observed in the cases regarding absence, duplication, bilateral asymmetry and tortuousity. Vernier calliper was used to note the size of the vessels and asymmetry of cerebral vessels of CW were assessed. It was not possible to measure the diameter of few specimens as size of fetal vessels was too small. In order to study the inter observer variability in measurements, we performed a paired "t" test between first and second measurement of the cases. Hypoplastic vessels were defined to be those with an external diameter of less than half the average. The observations were recorded as per the Performa. The results were tabulated and computed.
Results
Forty five fetal brains aged between 14 and 30 weeks were dissected (Fig.1.) In 71% of fetal brains, the configuration was normal as described in textbooks (Fig .4.) Variations in CW were noted in 12 (26.6%) cases which included duplication of anterior communicating artery (Fig.5.), hypoplasia (of anterior cerebral artery, posterior communicating artery, anterior communicating artery) in 4.4% cases respectively, and a single median anterior cerebral artery as shown in Table.1. Absence of A ComA was also observed (Fig.6.) In 2 specimens (4.4%), the two ACAs join to form a single, median ACA (azygous ACA) which later on splits into two ACA. (Fig.7). In all these cases, the continuity of the anastomosis is maintained. However in one specimen the AComA was absent, therefore the circle was termed as deficient (Fig.2, 7).
The variations affect the anterior half of the circle (22.2%) more than posterior half (4.4%). Also the communicating vessels showed more variations as compared to other main cerebral arteries (Fig.2.). The most commonly duplicated or fenestrated artery was AcomA (Fig5.), present in 4.4% of the cases in the present study (Table.1). The variations appeared more frequently on the left side than the right side.
Fetuses from age groups of 14-20+ wks mostly showed presence of all component vessels and less asymmetry. The diameter of P com A and PCA were apparently same and the so called "fetal or embryonic pattern" was not visible. Fetuses from age groups of 21-30 wks mostly showed presence of all component vessels and more asymmetry. The diameter of either PcomA or PCA was larger. It looked like the standard "fetal or adult pattern" (Fig.7). The transitional configuration was more prevalent (66.6%) in 14-20+ wks ; in later age groups the transitional configuration was observed in only 33.3% and in other specimens changed to either adult (40%) or fetal ( 35%) type.
Discussion
The CW starts developing around 4th week (24th day) of IUL and by 7th week (CRL 40mm), fully formed, closed CW can be visualised3.The standard textbook of Embryology 4 describe that initially the Internal carotid artery (ICA) divides into Anterior, Middle, and Posterior cerebral arteries. The latter extends backwards and divides into two branches- the medial branch joins the Basilar artery while the lateral extends to supply the occipital and temporal lobes. The growth of occipital lobes and brain stem provides the initial directive for the development of vertebro-basilar system. In due course of time blood from the basilar artery starts circulating in the medial branch and from there in the lateral branch supplying the temporal and occipital lobes. On account of hemodynamic factors, the proximal part of the embryonic PCA up to its division becomes smaller in calibre and is now called the posterior communicating artery (PComA).The medial and lateral branches of the original PCA are now named as proximal and distal segments of PCA arising from the basilar artery. The anterior communicating artery (AComA) appears in the human embryo at (CRL 18 mm) as a reticulated anastomosis between the two ACAs. At 24mm CR length, this network fuses to form a single AComA3.
Functional demands of the growing brain tissue govern the development of cerebral vasculature. Soon with the regression of some earlier vessels and appearance of new channels, a fully developed CW becomes functional at the end of 6-7weeks. As the cerebral vascular tree reaches its typical configuration, the multiple events that occur during the embryological stage might lead to a diverse spectrum of vascular anatomical variants1. Duplications and fenestrations of an artery can occur due to an incomplete fusion of arteries 2. This has been associated with the development of cerebral saccular aneurysm5.The theory behind the fact is that smooth muscle cells and collagen are decreased at the proximal and distal end of the fenestrated segment. Even in patients of subarachnoid haemorrhage, 95% of the cases are found to have fenestrations close to the site of hematoma6.
De Silva et al, (2010)7 concluded that there is a significant shift in the contribution of carotid versus basilar system. In older fetal brains, blood flow from internal carotid artery is more in PCA which changes to gradually more contribution from vertebra-basilar in the adult brains. Ardakami et al, (2008) 8 dissected 30 fetal and infantile brains observed hypoplasia of both communicating vessels similar to the present study. Absence of PComA was also reported by them, however in the present study absence of AComA was observed. They concluded that CW configuration in their samples was similar to adults as most of the cases were from infants not from fetuses.
Overbeek and Hilton,(1991) 9 in their exhaustive study on fetal brains observed three types of configuration in relation to the contribution of vertebro-basilar trunk. According to them, in very early gestational period (12-21wks), the most prevalent pattern was transitional (73%) in which all cerebral vessels are slender but of equal size. After 20-21 wks, the transitional phase decreased and changed into either fetal or adult type of configuration, similar to the present study (Fig.3.). According to literature2, there is a fast growth of the occipital lobes around 22-24th weeks of IU life. It demands an increase in blood supply. So to satisfy the increased functional demands, either the PComA ( branch of ICA) or the PCA (branch of basilar artery) enlarge and from that time an adult or fetal configuration may develop. Because of hemodynamic factors, the most favoured configuration is more enlargement of PCA i.e. adult configuration. Therefore the configuration of newly formed CW looks symmetrical and slender; this can be assumed as transitional phase; In later half of IU Life, this transitional phase will directly change into either fetal or in adult type of configuration.
According to Menshawi, (2015)2 the primary goal of the developing cerebral arterial system in the embryo is to supply blood to support the rapid and asymmetrical brain growth. As long as the flow is delivered, there should not be a deleterious immediate consequence to the brain or the vessel. In the long term, however, individuals with arterial configurations consisting of decreased collaterals routes (e.g., lack of PComA or AComA) or coexistence of branching arteries with discrepant diameters may be at a higher risk of developing atherosclerosis in areas with chronic low or oscillating blood flow.
In view of above mentioned literature, the anatomical variants of the fetal CW were classified and compared in the present study. The variations affect the anterior half of the circle more than posterior half and variations appeared more frequently on the right side. In this aspect, the present study differs from Krishnamurthy et al. (2006)10 where posterior part of circle showed larger number of variations. The variations appeared more frequently on the left side in the present study. Also the communicating vessels showed more variations as compared to other cerebral arteries.
Absence of A ComA was also reported by Critchley (1930)11 and Milenkovic et al,(1985)1 similar to the present study. All other component cerebral vessels were present in our fetal specimens; which is not in accordance with some other studies 12. In 2 specimens (4.4%), the two ACAs join to form a single, median ACA (azygous ACA) which later on splits into two ACA. De Vriese, (1905)13 and Menshawi, (2015)2 has also confirmed the presence of single median ACA in fetus as observed in the present study. According to them this vessel is present normally in primates. Therefore its presence in human brains can be attributed to its repetition in phylogeny.
A vessel can be classified as hypoplastic if the diameter is less than half the average, although it was not possible to measure the diameter in fetal brains, the vessels were not patent while injecting a coloured solution through the vessel. Hypoplasia of anterior cerebral artery was detected in 4.4% of the cases comparable to series of Milenkovic et al, (1985)1 (9.6%) and Riggs (1983)14 (7-28%).
Duplicated or fenestrated arteries are the second most common variants (after hypoplastic arteries) and are reportedly more frequent in the anterior circulation15. The most commonly duplicated or fenestrated artery was AcomA , present in 4.4% of the cases in the present study in contrast to incidence of 32% observed by Milenkovic et al,(1985)1, and Ardkani et al,(2008)8. Fenestration or duplications might be due to an incomplete fusion of arteries during development, however some authors believe fenestrations can also be induced by the trans-arterial course of a nerve, a bony structure or enlarged vasa vasorum along the path of the artery2. The presence of such variations in the study (hypoplasia / duplication /fenestration/ absence) in fetal brains lead to hypothesis that these are congenital in origin.
Fetuses from age groups of 14-20+ wks mostly showed presence of all component vessels and less asymmetry. The diameter of P com A and PCA were apparently same and the so called "fetal or embryonic pattern" was not visible. Fetuses from age groups of 21-30 wks mostly showed presence of all component vessels and more asymmetry. The diameter of either PcomA or PCA was larger. It looked like the standard "fetal or adult pattern" in the present study. These observations are in agreement with Padget DH,(1948)3, Overbeek and Hilton,(1991)7, Ardkani et al,(2008)8 and Menshawi et al,(2015)2, i.e. transitional configuration was more prevalent (60%) in 15-20+ wks ; in later age groups the transitional configuration was observed in only 25% and in brains from other fetuses changed to either adult (40%) or fetal ( 35%) type. In a similar study Raamt et al, (2006)16 has defined a term ' 'FTP' (Fetal Type Posterior CW) as full or partial. They have stated that in full FTP the P1 segment of PCA (i.e. contribution from basilar artery) is absent, thereby making the collateral flow entirely dependent on the anterior circulation of opposite side. The present study did not find even a single case with complete absence of P1 segment of PCA. Patients with fetal variation of CW are prone to develop vascular insufficiency as there exists an embryological defect in primary collateral circulation between anterior and posterior half of the circle17.
The number and type of variations in fetuses were lesser as compared to adults compared to a previous study done by the same author18 .One of the striking features of fetal CW was the absence of tortuous vessels which is so evident in adult brains. The literature does not throw any light on this aspect. It is assumed that development of tortuousity might be the result of ever expanding brain which increased the demand for more blood supply. Subsequently the increase in length of vessels will make it tortuous as because of anastomosis, the vessel is tied at both ends.
Asymmetry in the two halves of circle was observed in only 37.8% fetal brains on account of unilateral hypoplastic vessels.
There was no incidence of saccular aneurysm in fetal CW in the present study. None has been reported in literature also; thereby indicating that occurrence of saccular aneurysm is not congenital.
Conclusion
Abnormal pattern was observed in 26.6% fetal brains; however the CW was deficient in only 2.2% specimens. Variations in the posterior part of CW are the result of developmental modifications. Absence / duplication of AComA is also the persistence of early fetal formation of this artery by a plexus. The presence of a single median (fused) ACA in fetus could not be explained on embryological basis. It might be due to the persistence of an arrangement normally found in lower primates. Tortuousity of cerebral vessels was not observed in fetal brains. Asymmetry on the two sides was also observed in only 37.8% fetuses.
There was no evidence of presence of aneurysms in fetal brains. The cause of development of aneurysms later in life could be (i) congenital defect in the arterial wall (ii) hemodynamic stress (iii) combination of both.
It is concluded that variations existing early in fetal life could be a consequence of genetic factors whereas variations developing during adult life could be due to mechanical stress factors. Their recognition is important as some of these might be responsible for the development of cerebral atherosclerosis, stroke or even saccular aneurysm.
Conflict of interest
The authors declare that they have no conflict of interest concerning this article.
No funding was required for data collection, data analysis and data interpretation of the study.
Acknowledgement
Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are grateful to authors/editors/publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed.
Englishhttp://ijcrr.com/abstract.php?article_id=2324http://ijcrr.com/article_html.php?did=23241. Milenkovic Z, Vucetic R, Puzic M. Assymmetry and Anomalies of the Circle of Willis in fetal brain ,microsurgical study and functional remarks. Surg Neurol 1985;24:563-70.
2. Menshawi K, Mohr JP,and Gutierrez J. J Stroke 2015;17(2):144-58.
3. Padget DH. The circle of Willis: its embryology and anatomy. In: Dandy WE, Editor. Intracranial Arterial Aneurysms. New York. Comstock Publishing Company.1944.p:67-90
4. Hamilton WJ, Mossman HW. Cardiovascular system In : Hamilton, Boyd and Mossman's Human Embryology. Cambridge. The Williams and Wilkins Company 4th ed ;1972.p266-68.
5. LaBorde DV, Mason AM, Riley J, Dion JE, Barrow DL. Aneurysm of a duplicate middle cerebral artery. World Neurosurg. 2012;77:201.e1-201.e4.
6. Hudák I, Lenzsér G, Lunenkova V, Dóczi T. Cerebral arterial fenestrations: a common phenomenon in unexplained subarachnoid haemorrhage. Acta Neurochir (Wien). 2013;155:217-22.
7. De Silva KRD, Silva R, De Silva C, Gunasekera WSL, Dias P and Jayesekera RW. Comparison of the configuration of the posterior bifurcation of the posterior communicating artery between fetal and adult brains: A study of a Sri Lankan population. Ann Indian Acad Neurol.2010; 13(3):198-201.
8. Ardakani SK, Dadmehr M, Nejat F , Ansari S, Eftekhar B, and Tajik P et al. The cerebral arterial circle (Circulus Arteriosus Cerebri) A Anatomical study in fetus and infant samples.Pediatr Neurosurg. 2008;44:388-92.
9. Overbeeke JJV, Hillen B, Tulleken CAF. A comparative study of the circle of Willis in fetal and adult life. The configuration of the posterior bifurcation of the posterior communicating artery. J Anat.1991;176:45-54.
10. Krishnamurthy A,Rao CP, Naryana K, Nayak SR, Kumar SM, Surendran S. Circulus arteriosus cerebri: A study of variations in the fetal and adult human brains of south Indians. Morphologie. 2006; 90(290): 139-43
11 Critchley M. The anterior cerebral artery, and its syndromes. Brain. 1930;53:120-65.
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13. De Vriese B.Sur la sygnification morphologique des arteres cerebrales. Arch Biol. 1905;21:357-457.
14. Riggs HE, Rupp C. Variations in form of circle of Willis. Arch Neurol 1963;8:8-14.
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30HealthcareEffect of Gingival Crevicular Fluid Volume in Smokers
English3236Aishwarya A.S.English JaiganeshEnglishThe association between smoking and periodontal disease has been proven by many epidemiological evidences. This article reviews the recent studies and compiles the effect of smoking on Gingival crevicular fluid volume. Many studies support the fact that tobacco smoking affects the periodontium through different pathways like microcirculatory, host immune systems, connective tissue and bone mechanisms.
EnglishSmoking, Periodontal disease, Gingival crevicular fluidINTRODUCTION:
Periodontal disease is defined as inflammatory destruction of periodontal tissue and alveolar bone supporting the teeth. [5] Smoking is accepted as a risk factor for periodontal disease and detrimental to periodontal tissues. Many investigations suggest negative periodontal effects among smokers in comparison with non smokers. Smokers tend to have greater numbers of deeper periodontal pockets and probing depth. Studies show that cigarette smokers are 2 to 6 times more likely to develop severe periodontitis than in non-smokers [31,32]. Several epidemiological studies indicate that smoking has harmful effects on the response to non surgical and surgical procedures [1].
Paradoxically, smoking tends to mask the gingival inflammation by causing constriction of blood vessels of gingiva [20, 21]. Smoking is known to alter the host response including changes in vascular function, neutrophil activities, adhesion molecule expression, antibody production, release of cytokine inflammatory mediators [33,34].
Biological mechanism of periodontal disease is occurring due to the imbalance between bacterial virulence and host defence activity.[4]. The most possible mechanism that explains the relationship between smoking and periodontal disease is that smoking, an environmental factor, interacts with the host cells and affects the inflammatory changes [25]. These underlying mechanisms can be understood by linking findings of the previous epidemiological studies with in vitro studies.
Biological markers called as biomarkers which are indicative of the status of the disease and have been measured for several studies to reveal the mechanism of disease [27]. For example host immune inflammatory response systems have biomarkers like immune cells, immunoglobulins and metalloproteinase, cytokines, and adhesion molecules. These biomarkers are obtained limitedly particularly from non-diseased smokers. Thus commonly used specimens are saliva, blood serum, gingival crevicular fluid [26].
Gingival crevicular fluid is a fluid present in the sulcus or periodontal pocket between the surface of the tooth and the gingival epithelium [3]. Gingival crevicular fluid is a transudate as well as inflammatory exudates, produced by osmotic gradient containing low protein content. The amount of gingival fluid is greater when inflammation is present. Physiologically the fluid volume increases while masticating, brushing, ovulation etc[23]. Gingival crevicular fluid is a well known marker of gingival health status[1]. This study reviews articles relating the gingival fluid flow in smokers.
SEARCH ENTITY:
This article reviews based on the strategy, keywords like smoking and periodontitis, gingival fluid flow, Gingival crevicular fluid volume in smokers. Many articles reviewed were published in the last 5-7 years. This article constructs the relation between smoking and the gingival crevicular fluid flow rate by reviewing the previously conducted studies.
METHODS OF COLLECTING GINGIVAL CREVICULAR FLUID [7, 9]:
There are several methods to collect Gingival crevicular fluid
By using absorbent papers
Brill technique (intra sulcular method) -filter paper strip is placed inside the gingival sulcus until the resistance is felt.
Loe and Holm-Pedersen technique (extra sulcular technique) -filter paper strip is placed at the entrance of the gingival sulcus.
Collection of Gingival crevicular fluid using micropipettes
Micropipettes with standardized length and diameter and uses the capillary action to collect the fluid.
By using twisted threads
Pre weighed twisted threads are placed in the crevice and after placing, the threads are measured and measured.
Collecting Gingival crevicular fluid by using paper points
ESTIMATION OF AMOUNT OF GINGIVAL CREVICULAR FLUID:[7]
The amount of Gingival crevicular fluid collected in periopaper can be measured by several means
By weighing the absorbent medium:
The weight of the twisted threads are measured before and after placing in the gingival crevice and compared.
By measuring capillary fill of micropipettes
As the capillary tubes are marked with standardized length and diameter.
By staining with ninhydrin
The wetted area with Gingival crevicular fluid becomes more visible by staining with ninhydrin which can be then measured planimetrically on an enlarged photograph or with a microscope.
Electronic method
Amount of Gingival crevicular fluidcan be estimated by using an electronic device called as periotron.
COMPOSITION OF GINGIVAL CREVICULAR FLUID:
The contents of Gingival crevicular fluid are usually as a result of interaction between the bacterial biofilm and the periodontal tissues. It contains the products of tissue breakdown, inflammatory mediator and antibody produced against them[7]. From the hypothesis postulated by Pashley which suggested that the initial fluid produced represents interstitial fluid which appears in the crevice as the result of osmotic gradient. This initial, pre-inflammatory fluid is a transudate and on stimulation it becomes as an inflammatory exudates [8].
Composition of Gingival crevicular fluid
Enzymatic components
Host derived and other products
Bacteria derived
Non enzymatic components
Cellular components
Electrolytes
Organic components
Organic compounds: Glucose hexosamine and hexuronic acid are two main components of Gingival crevicular fluid, where the glucose level of blood do not coincide with the level of glucose in Gingival crevicular fluid, its concentration is three or four times greater than in serum level [6].
COMPOSITION OF CIGARETTES:
Tobacco smoke contains millions of noxious chemicals which comprises of gaseous phase and particulate phase. The gas phase contains carbon monoxide, ammonia, formaldehyde, hydrogen cyanide and 60 known carcinogen such as benzo(a)pyrine, dimethylnitrosamine. The particulate phase includes nicotine, benzene, and tar. Tar is in condensate form, sticky substance that stains the fingers and teeth yellow/brown. Nicotine an alkaloid found in tobacco leaves evaporates when the cigarette is lighted. It's quickly absorbed in the lungs and reaches the brain within 10-19 seconds. Nicotine is highly addictive and causes rise in blood pressure, increased heart rate and respiratory rate with peripheral vasoconstriction. Plasma and saliva cotinine concentration in smokers is approximately 300ng/ml and the urine concentration s approximately 1500 ng/ml. Non-smokers has plasma and saliva concentration
CHANGES IN GINGIVAL CREVICULAR FLUID COMPOSITION ASSOSCIATED WITH SMOKING:
Effect on microvasculature
Smaller number of vessels was observed in inflamed gingival tissues of smokers compared to non smokers. Smoking causes decrease in gingival blood flow and vascular conductance, leading to delay in healing and increases the risk of periodontal health [27,28]. Smoking causes inflammatory activation by inducing endothelial dysfunction which increases the cytokine adhesion molecules [14].
Host immune response
Smoking can affect the neutrophil count in blood in a dose-dependent manner [30]. Smoking reduces the proliferation of T lymphocytic cells and affects B cell function and antibody generation [30].
Connective tissue and bone metabolism:
Crevicular fluid in smokers has increased levels of Interleukins (IL 6, IL 8, IL 4), MMP, and free radicals. Gingival crevicular fluid has decreased levels interleukins (IL 1a), osteoprotegrin, prostaglandins, and gingival fibroblasts [4]
Viability of PMNs was significantly lower in light, moderate and heavy smokers compared to non-smokers. The ability of PMNs to phagocytose was significantly impaired in light, moderate and heavy smokers compared to non-smokers [13]. In periodontally diseased subjects the total amounts of IL-1β, IL-6 and IL-8 were significantly elevated as compared to healthy subjects, whereas IL-4 showed an inverse relationship to periodontal status and higher amounts were found in the healthy group[14].The levels of calprotectin, a leukocyte protein, in body fluids of patients with some inflammatory diseases are raised. Recently, we detected calprotectin in Gingival crevicular fluid and its concentrations in periodontal pockets were higher than those in healthy gingival crevices[16,17]. Investigations done by Jun-ichi Kido found the correlations between Gingival crevicular fluid calprotectin levels and clinical indicators (probing depth and bleeding on probing, BOP), and the IL-1β and PgE2 levels[14].
DISCUSSION:
Smoking is highly prevalent and can be considered an epidemic both developed and developing nations, smoking was higher in younger groups than older groups. Studies estimate that 1.3 billion people smoke per year worldwide on an average[6]. Smoking is detrimental to body tissues such as lungs, heart muscles, blood vessels, periodontium etc.
The periodontal probing depth and attachment loss were higher in smokers compared to non smokers [3]. Smoking tends to mask gingival inflammation by causing constriction of blood vessels of the gingiva [20,21,22]Smoking decreases oxygen tension in the gingival tissues. Smoking decreases tissue oxygen from from 65 7 to 44 ???????3 mmHg. Oxygen tension of 40-50 mmHg has increased risk of infection [9]. Vasoconstriction of blood vessels leads to reduced clinical signs and suppressed clinical expression of disease[11].A study done by Kemal et al showed that smoking significantly increased Gingival crevicular fluid flow/volume when compared to non-smokers. Purnima et al conducted a study with Gingival crevicular fluid, marginal and sub gingival plaque that revealed early acquisition and colonization of oral biofilm in smokers [12].
Cigarette smokers with periodontitis have increased periodontal destruction especially in palatal region of maxilla [35]. Study done by Xia Chen et al showed that 16% of smokers had higher chance of becoming edentulous within 10 years while only 0.3% of non-smokers become edentulous in the same period of time [15].
The research by Kaushal Luthra et al shows a clinically significant decrease in Gingival crevicular fluid volume in smokers when compared to non smokers and an increase in the volume of Gingival crevicular fluid 10 minutes post smoking[1]. Nicotine plays a major role in altering the gingival blood flow which subsequently correlates with the volume of Gingival crevicular fluid in smokers [2]. A similar result was reported by Morozumi et al[1]. There is an immediate increase in Gingival crevicular fluid volume five days after smoking cessation.
CONCLUSION:
The Gingival crevicular fluid volume is known to increase with the degree of inflammation. The increased Gingival crevicular fluid volume shows the presence of masked inflammation in smokers. Smoking alters the gingival blood flow which concomitantly relates to the Gingival crevicular fluid volume. This review concludes that the result may vary according to the pattern of smoking, methods of collection of Gingival crevicular fluid and sample size.
ACKNOWLEDGEMENT
The 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.
Englishhttp://ijcrr.com/abstract.php?article_id=2325http://ijcrr.com/article_html.php?did=23251. Ustun K, Nilgun A. Alptekin. The effect of tobacco smoking in gingival crevicular fluid. European Journal Dentistry, 1(4): 236-239, 2007.
2. Mokeem SA, Vellappally S , Preethanath RS , Hashem MI , Al-Kheraif AA , Anil S. Influence of Smoking on Clinical Parameters and Gingival Crevicular FluidVolume in Patients with Chronic Periodontitis, OHDM - Vol. 13 - No. 2,2014.
3. Luthra K, Grover HS, Aggarwal N, Luthra S. Smoking swings of gingival crevicular fluid secretion, Journal of Indian Society of Periodontology volume 16(1); 101-103, 2012.
4. Ojima, Hanioka: Destructive effects of smoking on molecular and genetic factors of periodontal disease. Tobacco induced diseases 2010 8: 4.
5. Sabrina C. Gomes, Flávia B. Piccinin, Rui V. Oppermann, Cristiano Susin, Rosemary Adriana C. Marcantonio. The effect of smoking on gingival crevicular fluid volume during the treatment of gingivitis.Acta Odontol. Latinoam Vol. 22 (3); 201-206,2009.
6.Newman, Michael G., Henry H. Takei, and Fermin A. Carranza. Carranza's Clinical Periodontology. Philadelphia: W.B. Saunders Co, 2002.
7. Textbook of periodontics by Jaiganesh, first edition.
8.Griffiths, Formation, collection and significance of Gingival crevicular fluid periodontal 2000:2003; volume 31,32-42.
9. Jensen J, Goodson WH, Hopf H, Hunt TK. Cigarette smoking decreases tissue oxygen. Arch Surg. 1991; 126(9):1131-1134.
10. Andrew J. Delina, origin and function of the cellular components in Gingival crevicular fluid, periodontal 2000:2003; volume 31,55-76.
11. Embery G, Waaddington R, Gingival Crevicular Fluid; Biomarkers of periodontal tissue Activity.Adv Dent Res 1994; 8(2):329-36.
12. Kumar PS, Matthews CR, Joshi V, de Jager M, Aspiras M. Tobacco Smoking Affects Bacterial Acquisition and Colonization in Oral Biofilms. Morrison RP, ed. Infection and Immunity. 2011;79(11):4730-4738.
13. Güntsch A, Erler M, Preshaw PM, Sigusch BW, Klinger G, Glockmann E. (2006), Effect of smoking on crevicular polymorphonuclear neutrophil function in periodontally healthy subjects. Journal of Periodontal Research, 41: 184-188.
14. Giannopoulou C, Kamma JJ, Mombelli A: Effect of inflammation, smoking and stress on gingival crevicular fluid cytokine level. J Clin Periodontol.2003, 30: 145-153.
15. Chen, X., Wolff L., AeppliD., Guo Z., Luan W, Baelum V. and Fejeskov O. (2001), Cigarette smoking, salivary/gingival crevicular fluid cotinine and periodontal status a 10-year longitudinal study. Journal of Clinical Periodontology, 28: 331-339.
16. Kido J, Nakamura, T, Kido R, Ohishi K, Yamauchi N, Kataoka M. and Nagata T(1999), Calprotectin in gingival crevicular fluid correlates with clinical and biochemical markers of periodontal disease. Journal of Clinical Periodontology, 26: 653-657.
17. Rai B, Kaur J, Anand S C, Laller K, The effect of smoking on gingival crevicular fluid levels of myeloperoxidase, Indian J Dent Res [serial online] 2010; 21:20-2.
18. Jonathan E, Smoking Heath effects and control, NEngl J Med 1985; 313:491-498
19. Preber H, Bergstrom J. The effect of non-surgical treatment on periodontal pockets in smokers and non smokers. J Clin Periodontol 1985; 13:319-23.
20. Bergstrom J, Floderus-Myrhed B. Cotwin control study of the relationship between smoking and some periodontal disease factors. Community Dent Oral Epidemiol 1983; 11:113-6.
21. Bergstrom J. Oral hygiene compliance and gingivitis expression in cigarette smokers. Scand J Dent Res. 1990; 98:497-503.
22. Baab DA, Oberg PA. The effect of cigarette smoking on gingival blood flow in humans. J Clin Periodontol. 1987; 14:418-24.
23. Preber H, Bergstrom J. occurrence of gingival bleeding in smoker and non smoker patients. Acta Odontol Scand. 1985:43:315-20.
24. Shapiro L, Goldman H, Bloom A. Sulcular exudates flow in gingival inflammation. J Periodontol. 1979; 50:301.
25. Palmer RM, Wilson RF, Hasan AS, Scott DA. Mechanism of action of environmental factors- tobacco smoking. J Clin Periodontol 2005, 32: 180-195.
26. Shield PG. Molecular epidemiology of smoking and lung cancer. Oncogene 2002, 21: 6870-6876.
27. Mavropoulos A, Aars H, Brodin P. Hyperaemic response to cigarette smoking in healthy gingiva. J Clin Periodontol 2003, 30: 214-221.
28. Mavropoulos A, Brodin P, Rosin CK, Aars H. Gingival blood flow in periodontitis patients before and after periodontal surgery assessed in smokers and non smokers. J Periodontol 2007, 78: 1774-1782.
29. Koundouros E, Odell E, Coward P, Wilson R, Palmer RM. Soluble adhesion molecules in serum of smokers and non smokers, with and without periodontitis. J Periodontal Res 1996, 31: 596-599.
30. Loos BG, Roos MTL, Schellekens PTA, Velden van der U, Miedema F. Lymphocyte numbers and function in relation to periodontitis and smoking. J Periodontol 2004, 75: 557-564.
31. Anand PS, Kamath KP, Shekar BR, Anil S. Relationship of smoking and smokeless tobacco use to tooth loss in a central Indian population. Oral Health and Preventive Dentistry. 2012; 10: 243-252.
32. Razali M, Palmer RM, Coward P, Wilson RF. A retrospective study of periodontal disease severity in smokers and non smokers. British Dental Journal. 2005; 198: 495-498; discussion 485.
33. Raulin LA, McPherson JC, 3rd, McQuade MJ, Hanson BS. The effect of nicotine on the attachment of human fibroblasts to glass and human root surfaces in vitro. Journal of Periodontology. 1998; 59: 318-325.
34. Ryder MI, Fujitaki R, Johnson G, Hyun W. Alterations of neutrophil oxidative burst by in vitro smoke exposure: implications for oral and systemic diseases. Annals of Periodontology. 1998; 3: 76-87.
35. Anil S. Study of the patterns of periodontal destruction in smokers with chronic periodontitis, Indian Journal of Dental Research, 2008; 19:124-128.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN2017September12HealthcareA Review on Lorcaserin - A Selective 5-HT Serotonin Receptor Agonist in Obesity Management
English3740Venkatesh P.English Venkhat Balaji B.R.English Venu Gopal K.English Vinodhini S.English Vinodhini T.English Vinodhini C.English Chitra K.EnglishLorcaserin is a novel anti-obesity agent. The study retrospect’s the pharmacokinetic effects, mode of action, adverse drug reactions and various uses of Lorcaserin. Lorcaserin is pro-opiomelanocortin neurons stimulator present in the nucleus of hypothalamus resulting in a peak melacortin-4 receptor activity, which results in satiety and decreased food intake. Though some side effects were reported, the potential benefits of Lorcaserin outweigh the risks. Serum drug monitoring is not required. Lorcaserin is a 5HT2C receptor agonist whose property may also be studied to treat anxiety, Alzheimer’s disease, depression and parkinsonism. Literature review was conducted to identify relevant studies. The study reviewed the pharmacokinetics, pharmacodynamics and clinical trials proposed so far on lorcaserin.
EnglishLorcaserin, 5-HT agonist, Anti-obesityIntroduction
Physical interventions such as exercise, diet and surgery, behavioural therapies, and pharmacological treatments are the approaches taken for the management of weight reduction in obese individuals. This may be done alone or in combination for greater efficiency.
Administration of anti-obesity drugs may lead to a reduction in the absorption of nutrients and appetite. It may also results in an increased satiety and energy expenditure. The better results were achieved with the pharmacotherapy for weight loss of about 2 to 7.9kg when compared to that treated with placebo.
5-HT is a monoamine neurotransmitter is mostly seen in the central serotogenic neurons and enterochromaffin cells with a broad spectrum of behavioural and physiological function. Hence, 5-HT receptor is considered as an anti-obesity drug target.
There is a wide range of 5-HT2c receptor modulating drugs having the ability to deal with a variety of conditions by changing the central serotogenic function. Such conditions are addiction, depression, anxiety, Alzheimer’s disease, parkinson’s disease and obesity1.
For the treatment of obesity only limited numbers of drugs are in use. In 1999, Orlistathas got approval by the Food and Drug Administration (FDA). Later in June 2012, a new drug Lorcaserin was approved and promoted for prescription by FDA. But In October 2012, it was rejected initially due to some cancer signal detection in animal studies. Finally, from further research such as BLOOM and BLOSSOM the drug was approved in the same year2.
Lorcaserinis chemically [1R]-8-chloro-2,3,4,5-tetrahydro-1-methyl-1H-3-benzapine and acts as a selective 5-hydroxy tryptamine (5-HT, serotonin)2c receptor agonist which is developed particularly to aim human appetite expression. Lorcaserin, a selective serotonin (5HT2c) receptor agonist is capable of suppressing appetite and food intake. Induction of this receptor gives rise to a number of reactions that finally stimulates the release of 2-melanocortin stimulating hormone, which acts on melanocortin-4-receptors to control appetite.3
Conclusion:
Lorcaserin is a novel anti-obesity agent. Since, it is a 5HT2C receptor agonist it may have potential in treating depression, anxiety, Alzheimer’s disease and Parkinsonism. The advantages of lorcaserin are high rate of renal excretion and minimal drug interaction. The common adverse drug reactions are nausea, dizziness, headache, vomiting, and cardiovalvulopathy. The attempt on review of Lorcaserin will pave the way for budding researchers to explore and fill the gaps in analytical methodology which are not so far reported and also useful for physicians and other health professionals to challenge their research on Lorcaserin.
Acknowledgement:
We sincerely thank the Management, Central Library, Publication oversight committee of Sri Ramachandra University for providing necessary scientific sources. 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.
Englishhttp://ijcrr.com/abstract.php?article_id=2326http://ijcrr.com/article_html.php?did=23261.Lisa L, Loannider Demos, Loretta Piccenna, John J Meneil. Pharmacotherapies for obesity; past, current and future therapies. Journal of Obesity 2011; Article ID 179674: 18pages.
2.Dick BS Bashim, A K Sharma, Navdeep Dahiya and Anjan Khadka. Lorcaserin; A novel anti-obesity drug. Journal of Pharmacology and Pharmacotherapeutics 2014; 5(2):175-9.
3. Jason C G, Halford and Joanne A Harold. Lorcaserin and the role of 5-HT2c agonist in the treatment of obesity. Clinical Medicine Reviews on Therapeutics 2011; (3): Page 347-354.
4. Lorcaserin. Clinical Pharmacology.[internet database].Gold Standard,inc.,2012.
Available at: http://www.clinicalpharmacology.comaccessed : November 12, 2012.
5.Lorcaserin. Lexi-drugs [database online].Lexi _comp, Inc: November 12, 2012.
6.Belviq [package insert].Woodcliff Lake, NJ: Eisai Inc., Ltd,: 2012
7.Lorcaserin. In: DRUGDEX System [Internent database]. Greenwood village, Colo: Thomson micromedex. Updated periodically.Accessed: Novemer 12: 2012.
8.Lorcaserin Facts and Comparisons [Internet Database]. Wolters Kluwer. Available at: http://online.factsandcomparison.com. Accessed: November 12, 2012.
9 Steven R Smith, Warren A Prossor, David , J Donahur, Michael E Morgan, Christen, MAndenon, et al. Lorcaserin (APD356), a selective 5-HT2c agonist, reduces body weight in obese men and women. Obesity 2009 March; 17(3): 494-593.
10. Steven R, Smith MD, Neil J. Weissman M, D Christen, M Anderson. Matilde S Multicenter, Placebo-controlled Trail of Lorcaserin For Weight Management.EnglJmed2010july 15, (363): 245-256.
11. Hurren KM, Berlie HD. Lorcaserin :An investigational serotonin 2C agonist for weight loss. American journal of health system pharm 2011 Nov 11; 68(21): 2029-37
12. Jun Goo Kang, Cheol–Young park. Anti –obesity Drugs :A Review about their effects and safety. Diabetes Metab 2012 Feb 17th; 36(1): 13-25.
13. ONeil PM, Smith SR , Weissman NJ , Fidler MC, Sanchez M ,Zhang J et al. Randomised placebo-controlled clinical trials of lorcaserin for weight loss in type 2 diabetes mellitus: the BLOOM –DM study. Journal of obesity 2012 July; 20(7): 1426-1436.
14.Chan EW, Hey Y, Chui CS, Wong AY, Lau WC, Wong IC. Efficacy and safety of lorcaserin in obese adults:a meta- analysis of 1-year randomized controlled trials (RCTS)and narrative review on short – term RCTS. Obes rev 2013May; 14(5): 383-392.
15. Joshua W, Fleming Katies MC, Clendon Daniel, M Riche. New obesity agents: Lorcaserin and pheneterimine / topiramate. Pharmacother 2013 June 25; 47(7): 1007-1016
16.Neil J Weismam, Matildesanchez, Gary G Koch, steven R smith, William R Shanahan, christen M Anderson. Echo cardiographic assessment of cardiac valvular regurgitation with lorcsaserin from analysis of 3 phase-3 clinical trials. Circulation: cardiovascular imaging 2013 July 16; 6:560-567.
17. Amal A Bajrai, Essam Ezzeldin, Khalid AAl-Rashood, Mohammad Raish and Muzaffar Iqbal. A validated UPLC-MS-MS Assay for the rapid determination of lorcaserin in plasma and brain tissue samples. Journal of analytical toxicology 2015 November 14; 40: 133-139.
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30HealthcareEmergency Obstetric Hysterectomy, Risk Factors, Indications and Outcome: A Retrospective Two Year Study
English4144Smita Somani BahetiEnglish Anjana VermaEnglish Medhavi SharmaEnglishBackground: Obstetric emergency hysterectomy means that emergency hysterectomy which is taken after drug therapy and conservative surgery fails to control blood loss from gravid uterus. It is the last resort to save mother’s life, and beside, the mother’s reproductive capability is sacrificed.
Objectives: To study the cases of peripartum hysterectomy over a period of 2 years from Jan 2015 – Dec 2016 in tertiary centre of Geetanjali Medical College and Hospital, Udaipur, to assess incidence, indication, risk factor and maternal outcome.Methods: 18 cases of emergency hysterectomies which were performed during the study period data were taken and were assessed.
Results: During the study period, incidence of obstetric hysterectomy is 0.52%. Most common indication for obstetric emergency hysterectomy is atonic PPH (27.77%). Hemorrhagic shock(61.11%) is most common complication followed by bladder injury(27.11%). Conclusion: Elderly gravida with IVF pregnancy, history of previous cesarean (with adherent placenta) and history of myomectomy are risk factors for peripartum hysterectomy. These cases should be dealt cautiouly and should be handled at tertiary centres.
EnglishObstetric emergency hysterectomy, Atonic postpartum hemorrhage, Hemorrhagic shock, Bladder injuryINTRODUCTION
Obstetric hysterectomy is a hysterectomy performed on a gravid uterus during pregnancy or in puerperium period. It was first done by Horatio Storer in 1869. Obstetric emergency hysterectomy means that emergency hysterectomy which is taken after drug therapy and conservative surgery fails to control blood loss from gravid uterus. On one hand it is the last resort to save mother's life, and beside, the mother's reproductive capability is sacrificed. Because of the increasing cesarean section (CS) rate world-wide and the concomitant rise in placenta previa and placenta accreta, the incidence of the emergency PH is rising1.
In third world countries, obstetric hemorrhage the uterine atony are the leading cause, of the maternal deaths, followed closely by ruptured uterus and uterine sepsis2. Emergency hysterectomy during normal vaginal deliveries, ectopic pregnancy or caesarian deliveries is performed when all other measures to control maternal hemorrhage have become futile. The commonest indication for emergency hysterectomy which are cited in the literature are uterine rupture and atonic uterus3.
Most of the times the operation is carried out when the condition of the patient is too critical to withstand the risks of anesthesia or surgery.
The purpose of our study was to know the incidence, indications, risk factors and the maternal profile and complications of the patients undergoing emergency hysterectomies at our tertiary level hospital which mainly caters to the rural and urban population.
AIMS AND OBJECTIVES:
The study was conducted with the following aims:
1. Incidence and indications of peripartum hysterectomy
2. Risk factors associated with peripartum hysterectomy
3. Maternal outcome
MATERIAL AND METHODS:
A retrospective study of 2 years from Jan 2015 upto Dec 2016 in tertiary centre of Geetanjali Medical College and Hospital, Udaipur Rajasthan, India was conducted. The data of 18 peripartum hysterectomies performed was collected and data was analyzed with special emphasis on indications, risk factors and maternal outcome.
OBSERVATIONS AND RESULTS
TABLE 1
INCIDENCES
Incidence of obstetrical hysterectomy in our study within 2 years of duration was 0.52%. In our study incidence of normal delivery was 40.01% and caesarean section was 59.98%. Maternal mortality was seen in (3/18) patients i.e (16.66%) cases.
INCIDENCES OF EMERGENCY HYSTRECTOMY
INCIDENCE
NUMBER
(N) = 3411
PERCENTAGE (%)
Incidence of Normal delivery
1365
40.01%
Incidence of Caesarean Delivery
2046
59.98%
Incidence of obstetric hysterectomy
18
0.52%
Incidence of obstetric hysterectomy followed vaginal delivery
5
0.14%
Incidence of obstetric hysterectomy followed caesarean section
7
0.20%
Incidence of obstetric hysterectomy due
to ectopic pregnancy
3
0.087%
TABLE 2
MATERNAL PROFILE
There was a high association of age see in our study. Majority women belonged to 26 -30 years of age .
MATERNAL AGE DISTRIBUTION IN EMERGENCY HYSTERECTOMY
AGE
(yrs.)
PARITY
0
1
2
3
4
5
TOTAL
46
2
-
-
-
-
-
2
Total
5
3
6
2
1
1
18
TABLE: 3
RISK FACTORS
The reason for this non -uniform distribution of parity with caesarean hysterectomy is due to presence of high risk factors, elderly IVF pregnancies, ectopic pregnancy and other confounding factors such as low socioeconomic status, poor general condition, massive hemorrhage and severe anemia.
RISK FACTORS ASSOCIATED WITH OBSTERICS HYSTERECTOMY
RISK FACTORS
NUMBER
PERCENTAGE(%)
Age > 35 years
11
61.11%
History of myomectomy
03
16.66%
History of previous LSCS (adherent placenta)
03
16.66%
IVF conception
05
27.77%
Multiple pregnancy
02
11.11%
Accidental hemorrhage
03
16.66%
Traumatic
01
5.55%
Where:
LSCS-lower segment Caesarean section
IVF in vitro fertilization
TABLE 4
INDICATIONS FOR OBSTETRIC HYSTERECTOMY
In our study most common indication for obstetrical hysterectomy was atonic PPH (27.77%) followed by rupture uterus 22.22%.
INDICATIONS FOR OBSTETRIC HYSTERECTOMY
INDICATIONS
NUMBER (N)
PERCENTAGE (%)
Traumatic PPH
1
5.55%
Atonic PPH
5
27.77%
Rupture uterus
4
22.22%
Ectopic pregnancy
3
16.66%
Placenta percreta
3
16.66%
Carcinoma in situ
1
5.55%
Molar gestation
1
5.55%
Total
18
100%
Where: (PPH : post partum hemorrhage)
TABLE 5
TYPE OF HYSTERECTOMY
In our study most common type of hysterectomy performed was total abdominal hysterectomy (66.66%) and 33.33% patients underwent subtotal hysterectomy.
TYPE OF HYSTERECTOMY
TYPE OF HYSTERECTOMY
NUMBER (N) N=18
PERCENTAGE (%)
Subtotal hysterectomy
6
33.33%
Total hysterectomy
12
66.66%
TABLE 6
POST OPERATIVE COMPLICATIONS
Amongst the post operative complications, the most common post operative complication in our study was haemorrhagic shock seen in (61.11%) patients followed by cases having bladder injury (27.77%), DIC (22.22%), acute renal failure(16.66%), paralytic ileus (16.66%) whereas, 16.66% patients had breast engorgement, wound infection (11.11%), 11.11% patients had septicemia and 16.66% was the documented maternal mortality rate.
POSTOPERATIVE COMPLICATIONS
CAUSES
NUMBER (N) N= 18
PERCENTAGE (%)
Breast engorgement
3
16.66%
Wound infection
2
11.11%
Bladder injury
5
27.11%
Septicemia
2
11.11%
Maternal mortality
3
16.66%
DIC
4
22.22%
Hemorrhagic Shock
11
61.11%
Paralytic Ileus
3
16.66%
Acute renal failure
3
16.66%
Where: DIC (disseminated intravascular coagulopathy)
DISCUSSION
Incidence of obstetrical hysterectomy in our study within 2 years of duration was 0.52% which was slightly higher to the studies conducted by Praneshwari et al4, Sturdee and Rushton5, Chew and Bishwas6, Gupta et al7 who reported an overall incidence of 0.0779%. 0.05%. 0.0392% and 0.26% each respectively. It may due to the fact that most of the deliveries at our tertiary care belong to high risk group and referral (referral cases high). (TABLE 1)
There was a high association of age see in our study but there was no significant difference seen in primi and multiparas in our study. Mean age of women who underwent obstetric hysterectomy at our centre was 35.44 years (TABLE 2). Study conducted by Najam R8 et al revealed 29% cases with parity >5. The reason for this non -uniform distribution of parity with caesarean hysterectomy is due to presence of high risk factors, elderly IVF pregnancies, ectopic pregnancy and other confounding factors such as low socioeconomic status, poor general condition, massive hemorrhage and severe anemia (TABLE 3).
In our study incidence of normal delivery was 40.01% and caesarean section was 59.98%. Whereas, incidence of obstetric hysterectomy followed by vaginal delivery was 0.14% and obstetrical hysterectomy followed by caesarean section was 0.20%. These results were slightly at a higher range as compared 0.0106%, 0.039% and 0.33%, 0.45% respectively reported by Praneshwari et al4and Pawar and Shroti et al9(TABLE 1).
In our study most common indication for obstetrical hysterectomy was atonic PPH (27.77%) followed by rupture uterus 22.22%, all ruptures are seen in previous scar uterus either scar due to myomectomy or due to Caesarean section no cases of rupture seen due to obstructed labour this could be due decreasing home delivery by untrained persons and promotion and practice of hospital deliveries. which was similar to the incidence found by Praneshwari et al4 (19.2%), Allahbadiya and Vaidya10 (16%), Kant Anita et al2(41.46%), Agashe and Marathe11 (60%) and Mantri et al13 (67.2%). Second most common indication in our study was rupture uterus (23.22%) which was similar to the study conducted by Praneshwari et al4(23%), Allahbadiya and Vaidya 10(20%) and Kant Anita2 (36.58%). In our study other indications seen were placental causes such as placenta increta and percreta(16.66%), ectopic pregnancy(16.66%), traumatic PPH (5.55%) and molar gestation (5.55%)(TABLE 4).
In our study maternal mortality was seen in (3/18) patients i.e (16.66%) cases. Similar results were found by Agashe and Marathe11 (14%). Whereas, Praneshwari et al4 found no maternal mortality in relation to obstetric hysterectomy.
In our study most common type of hysterectomy performed was total abdominal hysterectomy (66.66%). But subtotal hysterectomy is usually preffered as it is less time consuming surgery and it gives a better outcome in a moribund patient. But in indications like placenta previa and adherent placenta total abdominal hysterectomy is the ideal treatment as it removes the placental bed in the lower uterine segment. At our centre 33.33% patients underwent subtotal hysterectomy which was also seen by Praneshwari et al4 and Mrinalini et al12 (40%).(TABLE 5)
Amongst the post operative complications, the most common post operative complication in our study was haemorrhagic shock seen in (61.11% )followed by cases having bladder injury (27.77%), DIC (22.22%), acute renal failure(16.66%), paralytic ileus (16.66%) whereas, 16.66% patients had breast engorgement, wound infection (11.11%) and 11.11% patients had septicaemia. Whereas, Praneshwari et al4 found vesicovaginal fistula after subtotal hysterectomy which was done due to ruptured uterus which was followed by prolonged obstructed labor. Whereas, Kant Anita1 found post operative shock, pyrexia, paralytic ileus and wound infection as common post operative complications. They were mainly due to prolonged labour, intrauterine manipulations and sepsis. Nazam R8 reported 2 cases which had septic shock and 1 case in their study had DIC.(TABLE 6)
CONCLUSION
As life-saving procedure to deal with obstetric complication when medical and conservative surgical procedure fail emergency hysterectomy are performed. Elderlygravida with IVF pregnancy, history of previous LSCS(with adherent placenta) and history of myomectomy are risk factor for peripartum hystrectomy. These cases should be dealt cautiously and should be handled at tertiary centres. Impact of risk factors can be further studied by longerer duration of study. As a method of treatment it is a radical procedure, though it has a definite role in the management of life threatening obstetric hemorrhage or ruptured uterus. On one hand it is the last resort to save a mother's life, and on the other hand, the reproductive capability of a mother is sacrificed and leads to both surgical morbidity and psychological impact on women health.
Up gradation of the peripheral health centers and the timely referral of high risk parturients to higher centers can decline the rate of peripartum complications and improve maternal care and wellbeing. Emergency hysterectomy leads to psychological stress due to perceived loss off emininity, cessation of menstruation and reproductive ability. Psychological counselling and support therefore plays an important role in postoperative patients.
Acknowledgement:
Would like to thanks and 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.
Funding: No funding sources
Conflict of interest: None declared
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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241917EnglishN-0001November30HealthcareA Morphological Study on the Course, Branching Pattern and Termination of Peroneal Artery with a Note on the Nutrient Branch to Fibula
English4549C. Jeyanthi GnanadeepamEnglish Anjana T.S.R.English T. Preethi RamyaEnglish V. Raja PriyaEnglishIntroduction: The vessels of lower limb are prone for thrombosis in conditions such as Diabetes mellitus and Thromboangiitis obliterans. Apart from that, the fasciocutaneous flaps commonly employed in reconstruction procedures are based on the vessels of lower limb. In the wake of such increased importance, the present study is aimed at studying the course, branching pattern and termination of peroneal artery along with nutrient artery to fibula which is useful during fibular bone grafting procedures.
Materials and methods: The study was conducted in forty lower limb specimens from the Institute of Anatomy, Madras Medical College. The origin, course, branches of peroneal artery along with nutrient artery to tibia and fibula, perforating branches were observed in detail.
Results: The average distance of the origin of the peroneal artery from the lower border of popliteus was found to be 5.7 cm. The distance of origin of peroneal artery from the origin of posterior tibial artery was found to be 3.4 cm. The perforators arising from the peroneal artery were located predominantly in the distal two-quarters of the leg. The nutrient artery to fibula arose from the peroneal artery in all the specimens. It was two in number in 5 specimens (12.5%). There were three branches in two specimens (5%).
Conclusion: The knowledge of perforators is essential for raising skin flaps based on these branches. Since the flaps of these regions are considered to be reliable and easy to dissect, they are preferred even in primary centres also..
EnglishVessels, Lower limb, Perforator, Fasciocutaneous flap, Wound healingINTRODUCTION:
The arterial revascularization procedures have reinforced the significance of the course and variable branching patterns of vessels. It helps secure successful wound healing in the conduct of vascular and plastic surgery. The branching pattern of the arterial trunks of the lower limb gained importance following increased incidence of vascular diseases affecting lower limb especially in diabetics and smokers. The vessels are easily prone to intractable constriction following trauma. A fasciocutaneous flap brings additional blood supply to an infected area thereby promoting wound healing at the earliest.
A fasciocutaneous flap based on the branches of peroneal artery has been implicated in various reconstruction procedures. The present study aims to study the course, relations, branching pattern and termination of peroneal artery. Developmentally, it is the major artery of the leg. (Russel T Woodburne1961) The peroneal artery usually arises from the posterior tibial artery at about 2.5cm below the distal border of popliteus.(gray, Cunningham) and tendinous arch of Soleus( Keith L Moore, 1980). The artery passes laterally and is found to lie deep to the flexor hallucis longus to reach the back of lateral malleolus. (J D Boyd, 1956). Ben Pansky and E L House (1964), have described the peroneal artery as lying in a fibrous canal between tibialis posterior and flexor hallucis longus. Throughout its course, it lies in close relation to fibula and transverse crural intermuscular septum in the deep posterior compartment of leg. (Wesley S Moore, 2002).The artery usually terminates on the lateral surface of calcaneal tubercle by dividing into lateral calcaneal branches.( Parson Schaeffer,1953).
This vessel is exceedingly subject to variation and is rarely absent.3(George a Piersol). A high division of the popliteal artery may be associated with the origin of the peroneal artery from anterior tibial artery rather than the posterior tibial artery.2 (Trotter.M.1940). David Harvey(1990), has observed cases where the popliteal artery bifurcated into tibioperoneal trunk and anterior tibial artery. When the posterior tibial artery is diminished in size, the peroneal artery is relatively increased and conveys blood to the distal part of posterior tibial artery by means of a communicating branch.4(Frazer,1937). When the anterior tibial artery is small or absent, the peroneal artery enlarges and its perforating branch replaces the dorsalis pedis artery. ( Ronan O Rahily,1986).
In cases where the posterior tibial artery was absent, the peroneal artery was larger than usual and the vessel turned medially at the ankle joint level to resume the functions of posterior tibial artery. (J A Keen, 1961) and continued to form the lateral plantar artery( W H Hollinshead.1969). Piral T, Germain M,(1996) have noticed origin of anterior tibial artery from peroneal artery. Hence the peroneal artery is the most stable cruralartery.(Voboril R) for phylogenetic and embryologic reasons.
MATERIALS AND METHODS:
The present study was conducted in forty cadaveric lowerlimb specimens from the Instituite of Anatomy, Madras Medical College, Madras. An incision is made from the middle of popliteal fossa to the heel and then on to the level of middle toe. The skin, superficial fascia and deep fascia were reflected along with both heads of gastrocnemius and soleus. The popliteal artery was identified with its terminal branches, the anterior and posterior tibial artery. The peroneal artery is traced under cover of flexor hallucis longus and the following parameters were observed in relation to the artery.
Origin of peroneal artery.
Distance of origin of popliteal artery from the commencement.
Course of peroneal artery.
Origin of terminal branches- Medial and lateral plantar arteries.
Nutrient artery to tibia.
Nutrient artery to fibula.
RESULTS:
The various parameters related to the origin, distance of origin from the commencement of its posterior tibial artery or popliteal artery, nutrient branches to tibia and fibula were observed and the results were tabulated.
Table 1: ORIGIN OF PERONEAL ARTERY:
S no
Origin of peroneal artery
Number of specimens
Total number of specimens
Percentage(%)
1.
From posterior tibial artery
38
40
95
2.
Directly from popliteal artery
2
40
5
Table 2: DISTANCE OF ORIGIN OF PERONEAL ARTERY FROM THE LOWER BORDER OF POPLITEUS
Specimen number
Distance in cm
Specimen number
Distance in cm
1.
4.8
21.
5.0
2.
6.4
22.
6.8
3.
5.4
23.
4.8
4.
5.6
24.
6.4
5.
4.5
25.
5.0
6.
5.8
26.
5.5
7.
5.8
27.
6.2
8.
6.4
28.
5.8
9.
5.4
29.
6.6
10.
5.6
30.
5.4
11.
6.6
31.
4.6
12.
5.5
32.
4.4
13.
6.4
33.
5.2
14.
5.2
34.
4.8
15.
6.4
35.
4.7
16.
7.2
36.
5.4
17.
6.8
37.
6.3
18.
5.2
38.
5.2
19.
5.6
39.
6.3
20.
6.4
40.
5.5
The average distance of the origin of the peroneal artery from the lower border of popliteus was found to be 5.7 cm and ranging between 4.5cm to 7.2cm. The distance of origin of peroneal artery from the origin of posterior tibial artery was found to be 3.4 cm with the range of 2- 7.2cm.
Table 3: Course of peroneal artery:(Fig.1,2,3,4)
S. No
Course of peroneal artery
Numberof specimens (n =40)
Percentage (%)
1.
Normal course
34
85
2.
Enlarged and replaced posterior tibial artery distally
4
10
3.
Arising from popliteal artery with absence of posterior tibial artery.
2
5
BRANCHES:
CIRCUMFLEX FIBULAR ARTERY:
The circumflex fibular branch arose from the peroneal artery in two of the forty specimens, amounting its incidence to 5%. In 95% of specimens, the circumflex fibular artery was a branch of posterior tibial artery.
ORIGIN OF TERMINAL BRANCHES:
The terminal branches, medial and lateral plantar arteries arose from the peroneal artery in 15% of specimens (six lower limbs). In the remaining thirty four specimens (85%), the terminal branches were found to originate from the posterior tibial artery.
NUTRIENT ARTERY TO TIBIA(Fig.5):
In two specimens, where the posterior tibial artery was absent, the tibia received its nutrient artery from peroneal artery. In rest of the specimens, the tibia derived its nutrient artery from posterior tibial artery.
NUTRIENT ARTERY TO FIBULA(Fig.6):
The nutrient artery to fibula arose from the peroneal artery in all the specimens. It was two in number in 5 specimens (12.5%). There were three branches in two specimens (5%). There was a single nutrient artery in 33 lower limbs. The distance of the nutrient branch from the point of origin of the peroneal artery ranged between 7cm to 12cm. The origin was also measured from the styloid process of fibula and the distance varied between 13cm to 18cm.
PERFORATING BRANCHES(Fig.7):
The perforators arising from the peroneal artery varied between two to three in number. The perforators reached the anterior compartment of leg. The mean + standard deviation of the distance of the perforator from the lateral malleolus was 6.6 + 9.2cm.
DISCUSSION:
The observations made in relation to the origin, course and branches was compared with those of the results of the previous authors. The majority of studies including those by J.E.Frazer (1937), J Parsons Schaffer(1953), Russell T Woodburne(1961) have shown that the peroneal artery arose from the posterior tibial artery. G J Romanes(1964) had described the division of popliteal artery into anterior tibial and peroneal artery. In the present study, the peroneal artery originated directly from the popliteal artery with absence of posterior tibial artery in 6% of specimens. Henry Gray(1858), G J Romanes and Samendra Mitra (1973) had reported incidence of trifurcation of popliteal artery. In the present study, the trifurcation of popliteal artery into posterior tibial artery, peroneal artery and anterior tibial artery was not observed in any specimen.
The distance of the peroneal artery from the origin of posterior tibial artery was compared with those of the previous studies.
Table 4: Distance of peroneal artery from the origin of posterior tibial artery.
S NO
AUTHORS
DISTANCE (CM)
1.
Russell T Woodburne(1961)
2-3
2.
Keith.L.Moore (2005)
2-3
3.
BerishStrauch et al(1993)
3
4.
Harold Ellis (1980)
4
5.
Thomas Walmsley (1934)
5
6.
Henry Gray( 1858)
7-8
7.
Present study
2-7.2
The present study coincides with that of Henry Gray's observations with regards to the distance of origin of peroneal artery from the commencement of posterior tibial artery. The fibula can be used in bone grafting in avascular necrosis of hip bone, mandibular reconstruction etc. In such instances, the location and distance of nutrient artery to fibula will help surgeons during usage of fibula as a bone graft.
The perforators arsing from the peroneal artery were located predominantly in the distal two-quarters of the leg. The knowledge of perforators is essential for raising skin flaps based on these branches. Since the flaps of these regions are considered to be reliable and easy to dissect, they are preferred even in primary centres also. Sometimes these branches may be enlarged and may continue down as the dorsalis pedisartery. (Taser F).
CONCLUSION:
The revascularization procedures done as foot salvage surgeries require a detailed knowledge of the origin, course and relations of the blood vessels of lower limb. The peroneal artery being the predominant source of nutrient supply to fibula gains clinical importance especially during bone grafting procedures. The fibula is one of the bones commonly selected for bone grafting. Accessibility to these vessels is essential for the success of revascularization procedures. Hence the knowledge of the course, branching pattern and location of perforators enables surgeons and radiologists in achieving better results in surgeries or procedures involving the leg.
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.
Englishhttp://ijcrr.com/abstract.php?article_id=2328http://ijcrr.com/article_html.php?did=2328
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