028942   IMAMSARI M, PERTIWI-KOENTJORO M, NURHAYATI A P, ISDIANTONI, PRASETYO E N (Biology Dep, Faculty of Natural Science - Institut Teknologi Sepuluh Nopember (ITS), Surabaya - 60111, Indonesia, Email: endry@bio.its.ac.id) : In-vivo preliminary examination of Moringa oleifera leaves extract as antiaging candidate in swiss webster male mice (Mus musculus). Int J Pharm Sci Res 2018, 9(9), 3638-46.

Free radicals are molecules with an unpaired electron that very unstable and react quickly with other compound to gain stability. Once the process is started, it can be a cascade reaction resulting disruption of a living cell. The free radicals formed in our body are combated by antioxidants then interact with free radicals and terminate the chain reaction before vital molecules are damaged. Moringa oleifera leaf extract well known can be as a source of antioxidant since it contains flavanoids for reducing free radical. This study was design to produce a crude extract of dark green and bright green of M. oleifera leaves using different solvent to determine its antiaging effect in male mice Mus musculus Swiss Webster. The antioxidant capacity of the extract were carried out using TMAMQ (Tetramethoxy Azobizmethylene Quinon) and ABTS (2, 2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) method, while the antiaging activity were assessed by histological approach. The results showed that the highest antioxidant capacity was achieved by dark green M. oleifera leaves ethanolic extract followed by doubledestilated water (ddH₂0) and 95% n-hexane extracts. The antiaging activity were observed by level of M. musculus skin dermis thickness as 516.13 μm and more fibroblast cells 115.67 cells in the visual area of 157.883, 92 μm² of skin dermis. The result indicated that the colour of M. oleifera leaves and type of extraction solvent play important roles on the antioxidant capacity and antiaging activity.

3 illus, 1 table, 55 ref

030260   JUYAL G, SOOD A, MIDHA V, THELMA B K (Jawaharlal Nehru Univ, New Delhi - 110 067, Email: garimajuyal@gmail.com) : Genetics of ulcerative colitis: Putting into perspective the incremental gains from Indian studies. J Genet 2018, 97(5), 1493–507.

Ulcerative colitis (UC), one of the two clinical subtypes of inflammatory bowel disease is perceived as a potential ‘sleeping giant’ in the Indian subcontinent. Clinical manifestation is overall believed to be the same across ethnic groups but overwhelming genetics from large European and fewer non-European studies have revealed shared as well as unique disease susceptibly signatures between them, pointing to population specific differences at genomic and environmental levels. A systematic recount of the four major eras in UC genetics spanning earliest linkage analysis, cherry picked candidate gene association studies, unbiased genomewide association studies, their logical extension in trans-ethnic setting (Immunochip study), lastly whole exome sequencing efforts for rare variant burden; and lessons learnt thereof in context of genetically distinct Indian population was attempted in this review. Genetic heterogeneity manifesting at allelic/locus level across these approaches has been the consistent finding through the range of pan India studies. On the other hand, these salient findings also highlight the limitations of even the best of these genetic leads for prognostic/clinical application. The imminent need, therefore, for the UC research community to adopt newer approaches/tools with improved study design to (i) gain better insight into genetic/mechanistic basis of disease; (ii) identify biomarkers of immediate translational value; and (iii) develop new/alternate therapeutic options is emphasized at the end.

1 illus, 2 tables, 98 ref

030323   SRIGANESH R, PONNIAH R J (National Institute of Technology Tiruchirappalli, Tiruchirappalli - 620 015, Email: joseph@nitt.edu) : Genetics of language and its implications on language interventions. J Genet 2018, 97(5), 1485–91.

Genetic variation of language genes affect neurophysiology of brain and can thus influence the way people respond to environmental language input, leading to differences in terms of their response to environmental language learning. Conversely, language learning environment too can affect gene expressions through neuroepigenetic mechanisms, leading to increased interindividual differences. Further, language-related cognitive processes such as learning, working memory and perception; and language-related affective factors such as stress and positive emotion involve neuroplasticity, which is also epigenetically regulated. Language intervention methods must understand the extent and the type of difficulties, and must offer personalized learning and medical solutions. Medical intervention in terms of epigenetics and neurotransmitter regulation is proposed in addition to effective teaching methods to aid in effective language acquisition.

98 ref

030328   SUTO J-I, KOJIMA M (Institute of Agrobiological Sciences, Ibaraki 305-8634, Japan, Email: jsuto@affrc.go.jp) : Quantitative trait loci that determine plasma insulin levels in F₂ intercross populations produced from crosses between DDD/Sgn and C57BL/6J inbred mice. J Genet 2018, 97(5), 1413–20.

When compared to C57BL/6J (B6) mice, DDD/Sgn (DDD) mice has substantially higher plasma insulin levels in both sexes. In this study, we performed quantitative trait loci (QTL) mapping of plasma insulin levels in F₂ male mice produced by crosses between DDD and B6 mice. By single-QTL scans, we identified one significant QTL on chromosome 9. When body weight was included as an additive covariate, we identified two significant QTL on chromosomes 9 and 12; the latter coincided with a QTL that was previously identified in F₂ female mice produced by the same two strains. The inheritance mode and the direction of the allelic effect of QTL on chromosome 12 were similar in both sexes, but those on chromosome 9 differed between males and females, suggesting that the QTL on chromosome 9 was sex-specific. Based on phenotypic correlations of plasma insulin levels with body weight and plasma levels of total cholesterol, triglyceride and testosterone, we subsequently assessed whether these insulin QTL explain the variation in other metabolic traits by using a point-wise significance threshold of P = 0.05. QTL on chromosome 12 had no significant effect on any trait. In contrast, QTL on chromosome 9 had significant effects on body weight and total cholesterol level. We postulate that Gpr68 and Cyp19a1 are plausible candidate genes for QTL on chromosomes 12 and 9, respectively. These findings provide insight into the genetic mechanisms underlying insulin metabolism.

4 illus, 6 tables, 14 ref

030292   OZBAYER C, KURT H, KEBAPCI M N, GUNES H V, COLAK E, DEGIRMENCI I (Medical Biology Dep, Eskisehir Osmangazi Univ, Eskisehir 26480 , Turkey, Email: hkurtayda@gmail.com) : The genetic variants of solute carrier family 11 member 2 gene and risk of developing type-2 diabetes. J Genet 2018, 97(5), 1407–12.

Type-2 diabetes (T2DM) is a metabolic disorder characterized by long-term insulin resistance, impaired insulin secretion from β-cells, and loss of beta cell mass and function. Inflammation and oxidative stress play a key role in the development of diabetes and are associated with insulin resistance. Notably, recent studies have demonstrated an association between body iron stores, insulin resistance and T2DM. Free iron, a powerful pro-oxidant molecule, is involved in oxidative stress, lipid peroxidation and endothelial dysfunction via its ability to generate free radicals. Specifically, the accumulation of iron in beta cells triggers oxidative stress and DNA damage, which have been reported to be associated with β-cell death and apoptosis. Solute carrier family-11 member-2 (SLC11A2) functions to transport ferrous iron and some divalent metal ions throughout the plasma membrane and across endosomal membranes. Functional polymorphisms in the SLC11A2 gene have been reported to cause excess storage of iron, resulting in iron overload. In this study, we evaluated the association between T2DM and SLC11A2 gene variants IVS4+44C/A, 1303C/A and 1254T/C by performing PCR-RFLP analysis on 100 T2DM patients and 100 healthy subjects. PCR products were digested with MnlI, MboI and SfanI restriction endonucleases and the products were then separated by 3 % agarose gel electrophoresis. The genotype frequencies of the 1254T/C and 1303C/A SLC11A2 gene variants did not differ between healthy controls and T2DM patients (P > 0.05). But, in recessive model (P = 0.037) and homozygous CC genotype (P = 0.030) for IVS4+44C/A showed significant correlation with T2DM risk. It is thought that presence of C allele of IVS4+44C/A plays pathological roles.

3 illus, 4 tables, 22 ref

030246   DU J, LI Y, SUN X, YU L, JIANG H, CAO Q, SHANG J, SUN M, LIU Y, SHU K (Sichuan Agricultural Univ, Chengdu 611130, China, Email: junbodu@hotmail.com) : Characterization of a splice variant of soybean ERECTA devoid of an intracellular kinase domain in response to shade stress. J Genet 2018, 97(5), 1353–61.

The receptor-like kinase ERECTA (ER) plays vital roles in plant adaptation under environmental stress including shade avoidance in Arabidopsis. In our previous study, we identified four ER paralogues in soybean (GmERs) that showed high similarities to Arabidopsis ER. Each GmER was predicted to generate diverse alternative splicing variants. However, whether soybean GmERs contribute to shade avoidance is unknown. Here we report our characterization of GmERs in response to shading. Promoter::GUS staining analysis shows that expression of GmER paralogous genes was differentially induced under shade stress. Further analyses show that GmERa.1 and GmERa.2 exhibit a larger distinction in length than the other GmER variants. GmERa.2 has the shortest length of amino acid with only 15 leucine-rich repeats which is the part of the extracellular domain of GmERa.1. Overexpression of GmERa.2 fully rescued the hypocotyl length, leaf area and petiole length, and the sensitivity of the hypocotyl of Arabidopsis mutant er-3 to shading, suggesting that the truncated extracellular domain of GmERa might contribute importantly to shade avoidance.

5 illus, 33 ref

030334   TURNER A N , ANDERSEN R S, BOOKOUT I E, BRASHEAR L N, DAVIS J C, GAHAN D M, GOTHAM J P, HIJAZ B A, KAUSHIK A S, MCGILL J B, et al. (Genetics Dep, Alabama at Birmingham Univ, Birmingham, AL 35294, USA, Email: challa.anilkumar@gmail.com) : Analysis of novel domain-specific mutations in the zebrafish ndr2/cyclops gene generated using CRISPR-Cas9 RNPs. J Genet 2018, 97(5), 1315–25.

Nodal-related protein (ndr2) is a member of the transforming growth factor type β superfamily of factors and is required for ventral midline patterning of the embryonic central nervous system in zebrafish. In humans, mutations in the gene encoding nodal cause holoprosencephaly and heterotaxy. Mutations in the ndr2 gene in the zebrafish (Danio rerio) lead to similar phenotypes, including loss of the medial floor plate, severe deficits in ventral forebrain development and cyclopia. Alleles of the ndr2 gene have been useful in studying patterning of ventral structures of the central nervous system. Fifteen different ndr2 alleles have been reported in zebrafish, of which eight were generated using chemical mutagenesis, four were radiation-induced and the remaining alleles were obtained via random insertion, gene targeting (TALEN) or unknown methods. Therefore, most mutation sites were random and could not be predicted a priori. Using the CRISPR-Cas9 system from Streptococcus pyogenes, we targeted distinct regions in all three exons of zebrafish ndr2 and observed cyclopia in the injected (G₀) embryos. We show that the use of sgRNA-Cas9 ribonucleoprotein (RNP) complexes can cause penetrant cyclopic phenotypes in injected (G₀) embryos. Targeted polymerase chain reaction amplicon analysis using Sanger sequencing showed that most of the alleles had small indels resulting in frameshifts. The sequence information correlates with the loss of ndr2 activity. In this study, we validate multiple CRISPR targets using an in vitro nuclease assay and in vivo analysis using embryos. We describe one specific mutant allele resulting in the loss of conserved terminal cysteine-coding sequences. This study is another demonstration of the utility of the CRISPR-Cas9 system in generating domain-specific mutations and provides further insights into the structure–function of the ndr2 gene.

4 illus, 1 table, 39 ref

030341   WANG S, PENG M C, CHEN X, LIU C Y, CHEN Y, LIU X D, ZHOU R B (Hunan Univ of Chinese Medicine, Changsha, China, Email: paeonia_dd@126.com) : Molecular cloning and spatiotemporal expression of APETALA1-like gene in Lonicera macranthoides. J Genet 2018, 97(5), 1281–8.

APETALA1 (AP1), a floral meristem identity gene controls the flowering time and floral transition, and plays an important role in inflorescence and floral organ development. The full-length cDNA for AP1 was obtained by rapid amplification of the cDNA ends (RACE) so that the roles of AP1 in Lonicera macranthoides (Lm-AP1) could be better understood. AP1 (accession number in GenBank: MF418642) consisted of a 729-bp open reading frame encoding a protein that contained 242 amino acids, had a deduced molecular mass of 27.9919 kDa and a theoretical isoelectric point of 8.75. No signal peptide or transmembrane domains were detected in the sequences located in the nucleus, but it contained conserved sequences for MADS and the K-box. In the secondary structure, the α-helix accounts for 60.74 %, the β-turn 3.72 %. The real-time polymerase chain reaction revealed that AP1 was more highly expressed in flowers, especially at the fourth flowering stage, which implied that it may play a role in flower development. Other L. macranthoides organs, such as stems and leaves, also expressed AP1. This research provided the basis for further analysis of the AP1 functional mechanism during L. macranthoides development.

6 illus, 31 ref

030330   TAUNK J, RANI A, YADAV N R, YADAV D V, YADAV R C, RAJ K, KUMAR R, YADAV H P (Molecular Biology Dep, Chaudhary Charan Singh Haryana Agricultural Univ, Hisar - 125 004, Email: nryadav58@gmail.com) : Molecular breeding of ameliorating commercial pearl millet hybrid for downy mildew resistance. J Genet 2018, 97(5), 1241–51.

Downy mildew (DM) caused by Sclerospora graminicola is the most calamitous disease of pearl millet. Therefore, for introgression of DM resistance (DMR) in HHB 197 (MH-1302), an elite pearl millet hybrid, a marker-assisted breeding was undertaken by targeting three DMR loci on linkage groups (LGs) 1, 2 and 4. Breeding programme was initiated by crossing HBL 11 (DM susceptible), male parent of HHB 197 hybrid with ICMP 451 (DM-resistant) to produce true F₁ plants. By conducting three rounds of backcrossing and selection, BC₃F₁ lines were generated. Foreground selection was employed using six polymorphic simple sequence repeat (SSR) markers of the 18 total selected markers. Four of these markers were linked to LG 1, five to LG 2 and nine to LG 4. Background selection was performed in BC₃F₁ generation using 33 polymorphic SSR markers of a total of 56 evenly spread SSR markers in the pearl millet genome to check recovery of recurrent parent genome. On the basis of genotypic selection (foreground as well as background) using selected SSR markers, agronomic performance in field and DM screening in greenhouse; 10 improved HBL 11 lines were selected and crossed with ICMA 97111 to produce DM-resistant HHB 197 hybrid versions. Six putatively improved HHB 197 hybrids were successfully tested in first year trials at Hisar and Bawal locations of Haryana and two selected versions with higher yield and zero DM incidence will be further tested in multilocation trials.

4 illus, 5 tables, 36 ref

030247   EZZIDI I, MTIRAOUI N, ALI M E M, MASOUDI A A, DUHIER F A (Monastir Univ, Monastir, Tunisia, Email: iezzidi@ut.edu.sa) : Impact of variants on type-2 diabetes risk genes identified through genomewide association studies in polycystic ovary syndrome: A case-control study. J Genet 2018, 97(5), 1213–23.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder in females, and is associated with altered metabolic processes in particular insulin resistance and diabetes mellitus. PCOS shares with type-2 diabetes (T2D) a number of features, including beta cell dysfunction, impaired glucose tolerance and dyslipidaemia. Recently, genomewide association studies (GWAS) have reported a number of genes with reproducible associations and susceptibilities to T2D. To address this, we examined the association between the T2D GWAS candidate genes (CDKAL1, CDKN2B, COL8A1, HHEX, IGF2BP2, KCNJ1, KCNQ1 and SLC30A8) and PCOS in Saudi women. A case–control study, includes 162 cases and 162 controls was enrolled. Genotyping was carried out by the allelic discrimination method. Our results showed that the variants including rs792837 of COL8A1, rs61873498 of KCNQ1 and rs13266634 of SLC30A8 genes to be significantly more frequent in PCOS patients than in controls. Our results suggest that COL8A1, KCNQ1 and SLC30A8, which are previously identified through GWAS as T2D-associated genes, are associated with PCOS.

4 tables, 34 ref

030298   PÉREZ L O, CANCELARICH N L, AGUILAR S, BASSO N G, MARANI M M (Instituto Patagónico para el Estudio de Ecosistemas Continentales, Puerto Madryn, Argentina, Email: mmarani@cenpat-conicet.gob.ar) : Genetic analysis of signal peptides in amphibian antimicrobial secretions. J Genet 2018, 97(5), 1205–12.

Amphibian secretion is an important source of bioactive molecules that naturally protect the skin against noxious microorganisms. Collectively called antimicrobial peptides (AMPs), these molecules have a wide spectrum of action, targeting viruses, bacteria and fungi. Like many membrane and secreted proteins, AMPs have cleavable signal sequences that mediate and translocate the nascent polypeptide chains into the endoplasmic reticulum. Although it is accepted that the signal peptides (SPs) are simple and interchangeable, there is neither sequence nor structure that is conserved among all gene families. They derived from a common ancestor but developed different traits as they adapt to distinct environmental pressures. The aim of this study was to provide an overview of the diversity of SPs of the frog, taking into account reported cDNA sequences and the evolutionary relationship among them. We analysed more than 2000 records that reported the relative abundance, diversity and evolutionary divergence based on the peptide signals of frog AMPs. We conclude that the physical properties of the sequence are more important than the specific peptides in AMP SPs. Since there is significant overlapping among related genera, differences in secretion from different peptide types should be regulated by additional levels, such as posttranscriptional modifications or 5-UTR sequences.

4 illus, 1 table, 34 ref

030254   IDNURM A (The Univ of Melbourne, Melbourne, VIC 3010, Australia, Email: alexander.idnurm@unimelb.edu.au) : Mystique of Phycomyces blakesleeanus is a peculiar mitochondrial genetic element that is highly variable in DNA sequence while subjected to strong negative selection. J Genet 2018, 97(5), 1195–204.

A DNA region in the mitochondrial genome of the fungus Phycomyces blakesleeanus (Mucorales, Mucoromycota) was characterized in a population of wild-type strains. The region encodes a predicted protein similar to the reverse transcriptases encoded by mitochondrial retroplasmids of Neurospora species and other Sordariomycetes (Ascomycota), but is uncommon in other fungi. DNA sequences of this element, named mystique, are highly variable between the strains, having greater than 2.5% divergence, yet most of the nucleotide differences fall in codon positions that do not change the amino acid sequence. The high proportion of polymorphisms coupled to the rarity of nonsynonymous changes suggests that mystique is subject to counteracting forces of hypermutation and purifying selection. However, while evidence for negative selection may infer that the element provides a fitness benefit, some strains of P. blakesleeanus do not have the element and grow equivalently well as those strains with it. A mechanism to explain the variability between the mystique alleles is proposed, of error-prone replication through an RNA intermediate, reverse transcription and reintegration of the element into the mitochondrial genome.

7 illus, 1 table, 34 ref

030227   AZIZ A A A, SALLEH M S M, MOHAMAD I, BHAVARAJU V M K, YAHYA M M, ZAKARIA A D, GAN S H, ANKATHIL R (Human Genome Centre, Kelantan, Malaysia, Email: rankathil@hotmail.com) : Single-nucleotide polymorphisms and mRNA expression of CYP1B1 influence treatment response in triple negative breast cancer patients undergoing chemotherapy. J Genet 2018, 97(5), 1185–94.

Triple negative breast cancer (TNBC) is typically associated with poor and interindividual variability in treatment response. Cytochrome P450 family 1 subfamily B1 (CYP1B1) is a metabolizing enzyme, involved in the biotransformation of xenobiotics and anticancer drugs. We hypothesized that, single-nucleotide polymorphisms (SNPs), CYP1B1 142 C>G, 4326 C>G and 4360 A>G, and CYP1B1 mRNA expression might be potential biomarkers for prediction of treatment response in TNBC patients. CYP1B1 SNPs genotyping (76 TNBC patients) was performed using allele-specific polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism methods and mRNA expression of CYP1B1 (41 formalin-fixed paraffin embedded blocks) was quantified using quantitative reverse transcription PCR. Homozygous variant genotype (GG) and variant allele (G) of CYP1B1 4326 C>G polymorphism showed significantly higher risk for development of resistance to chemotherapy with adjusted odds ratio (OR): 6.802 and 3.010, respectively. Whereas, CYP1B1 142 CG heterozygous genotype showed significant association with good treatment response with adjusted OR: 0.199. CYP1B1 142C-4326G haplotype was associated with higher risk for chemoresistance with OR: 2.579. Expression analysis revealed that the relative expression of CYP1B1 was downregulated (0.592) in cancerous tissue compared with normal adjacent tissues. When analysed for association with chemotherapy response, CYP1B1 expression was found to be significantly upregulated (3.256) in cancerous tissues of patients who did not respond as opposed to those of patients who showed response to chemotherapy. Our findings suggest that SNPs together with mRNA expression of CYP1B1 may be useful biomarkers to predict chemotherapy response in TNBC patients.

3 illus, 5 tables, 51 ref

030282   MÉNDEZ-RODRÍGUEZ A, LÓPEZ-WILCHIS R, DÍAZ A S, JUSTE J, RÍO-PORTILLA M A D, GUEVARA-CHUMACERO L M (Biología Dep, Autónoma Metropolitana Iztapalapa Univ, Mexico, Email: rlw@xanum.uam.mx) : Isolation and characterization of microsatellite marker loci in the Wagner's mustached bat Pteronotus psilotis (Chiroptera: Mormoopidae) and cross-amplification in other related species. J Genet 2018, 97(5), 1179–83.

Pteronotus psilotis, a mormoopid bat, is an insectivorous, gregarious and strict cave-dwelling species that is found areas between the sea level and an elevation of about 1000 masl. This species is present in diverse habitats ranging from rain forest to dry deciduous forest. Nine microsatellite loci were developed for Wagner’s mustached bat, Pteronotus psilotis using the next-generation sequencing approach, and their utility for population genetics studies was assessed. All loci were polymorphic (7–15 alleles) and characterized in 30 individuals from three P. psilotis populations, with the levels of observed and expected heterozygosity ranging from 0.280 to 0.867 and 0.584 to 0.842, respectively. One locus showed significant departures from Hardy–Weinberg expectations after Bonferroni correction. Cross-amplification in 11 other bat species was tested, for which eight microsatellites were successfully amplified, and of these seven were polymorphic. The development of these new microsatellite loci will contribute to investigations of genetic population structure, genetic diversity and gene flow in P. psilotis populations, as well as in other closely related bat species.

1 illus, 3 tables, 26 ref

030250   GREJTAKOVA D, GABRIKOVA-DOJCAKOVA D, BORONOVA I, KYJOVSKA L, HUBCEJOVA J, FECENKOVA M, ZIGOVA M, PRIGANC M, BERNASOVSKA J (Biology Dep, Presov Univ, Presov, Slovakia, Email: daniela.grejtakova@gmail.com) : WNT10A variants in relation to nonsyndromic hypodontia in eastern Slovak population. J Genet 2018, 97(5), 1169–77.

Nonsyndromic hypodontia is a congenital absence of less than six permanent teeth, with a most common subtype maxillary lateral incisor agenesis (MLIA). Mutations in several genes have been described in severe tooth agenesis. The aim of this study was to search for the variants in wingless-type MMTV-integration site family member (WNT10A), paired box 9 (PAX9) and axis inhibitor 2 (AXIN2) genes, and investigate their potential role in the pathogenesis of non-syndromic hypodontia. Clinical examination and panoramic radiograph were performed in the cohort of 60 unrelated Slovak patients of Caucasian origin with nonsyndromic hypodontia including 37 MLIA cases and 48 healthy controls. Genomic DNA was isolated from buccal swabs and Sanger sequencing of WNT10A, PAX9 and AXIN2 was performed. Altogether, we identified 23 single-nucleotide variants, of which five were novel. We have found three rare nonsynonymous variants in WNT10A (p.Gly165Arg; p.Gly213Ser and p.Phe228Ile) in eight (13.33 %) of 60 patients. Analysis showed potentially damaged WNT10A variant p.Phe228Ile predominantly occurred only in MLIA patients, and with a dominant form of tooth agenesis (odds ratio (OR_dom) = 9.841; P = 0.045; 95 % confidence interval (CI) 0.492–196.701; O_Rrec = 0.773; P = 1.000; 95 % CI 0.015–39.877). In addition, the WNT10A variant p.Phe228Ile showed a trend associated with familial nonsyndromic hypodontia (P = 0.024; OR = 1.20; 95% CI 0.97–1.48). After Bonferroni correction, these effects remained with borderline tendencies. Using a 3D WNT10A protein model, we demonstrated that the variant Phe228Ile changes the protein secondary structure. In PAX9 and AXIN2, common variants were detected. Our findings suggest that the identified WNT10A variant p.Phe228Ile could represent risk for the inherited nonsyndromic hypodontia underlying MLIA. However, further study in different populations is required.

3 illus, 5 tables, 38 ref

030231   CHANG C, XI L, ZHANG J, ZHAO W, LIU Z, GUO J, XU C (Histology and Embryology, Xinxiang Medical Univ, Xinxiang 453007, China, Email: zhangjihong@xxmu.edu.cn) : Roles of Cyclin A, Myc, Jun and Ppm1l in tumourigenic transformation of NIH3T3 cell. J Genet 2018, 97(5), 1155–68.

To analyse the mechanism of tumourigenic transformation of NIH3T3 cells at the transcriptional level, we used cancerogen 3-methylcholanthrene (3-MCA) and cancerogenic substance phorbol-12-myristate-13-acetate (PMA) to transform NIH3T3 cells and the assessment of transformation was performed using Giemsa staining and methylcellulose colony formation assay. Changes in gene expression profile were detected by Mouse Genome 430 2.0 microarray; and quantitative real-time polymerase chain reaction and Western blotting were used to verify the expression changes of mRNAs and proteins, respectively. With the aid of bioinformatics method, five signalling pathways were identified to participate in different stages of NIH3T3 cell transformation. Further, our study suggested that oncogenes Cyclin A, Myc, Jun and the tumour suppressor gene Ppm1l may play important roles in these pathways.

6 illus, 4 tables, 40 ref

030281   MEMON S, LI G, XIONG H, WANG L, LIU X, YUAN M, DENG W, XI D (Yunnan Agricultural Univ, Kunming 650201, China, Email: 2522935343@qq.com) : Deletion/insertion polymorphisms of the prion protein gene (PRNP) in Gayal (Bos frontalis). J Genet 2018, 97(5), 1131–8.

Resistance to fatal disease bovine spongiform encephalopathy (BSE), due to misfolded prion protein in cattle, is associated with a 23-bp indel polymorphism in the putative promoter and a 12-bp indel in intron 1 of the PRNP gene. Gayal (Bos frontalis) is an important semiwild bovid species and of great conservation concern, but till today these indel polymorphisms have not been evaluated in gayals. Therefore, we collected 225 samples of gayals and evaluated the genetic indel polymorphism in the two regions of this PRNP gene. The results revealed high allelic frequencies of insertions at these indel sites: 0.909 and 0.667 for, respectively, the 23 bp and 12 bp indels, both also with significant genotype frequencies (χ²: 9.81; 23 bp and χ²: 43.56; 12 bp). At the same time, the haplotype data showed indel polymorphisms with extremely low deletion (0.01) in both regions of the PRNP gene. We compared these data with those reported for healthy and BSE-affected cattle (Bos taurus) breeds from two European countries, Germany and Switzerland, and significant difference (P < 0.001) was observed between BSE-affected as well as the healthy cattle. Further, our data were also extensively compared with previous reports on BSE and highly significant (P < 0.001) outcomes were observed. This result suggested negligible genetic susceptibility to BSE in gayals. To the best of our knowledge, this study is the first comprehensive deciphering information about the PRNP indel polymorphisms of 23 bp and 12 bp in gayals, a semiwild species of China.

1 illus, 4 tables, 47 ref

030348   ZHANG W, WANG S-Y, DENG S-Y, GAO L, YANG L-W, LIU X-N, SHI G-Q (Xinjiang Agricultural Professional Technological Coll, Changji 831100, China, Email: wangshiyinxjnzy@163.com) : MiR-27b promotes sheep skeletal muscle satellite cell proliferation by targeting myostatin gene. J Genet 2018, 97(5), 1107–17.

To investigate the role of miR-27b in sheep skeletal muscle development, here we first cloned the sequence of sheep pre-miR-27b, then further investigated its expression pattern in sheep skeletal muscle in vivo, the relationship of miR-27b expression and sheep skeletal muscle satellite cell proliferation and differentiation in vitro, and then finally confirmed its target gene during this development process. MiR-27b sequence, especially its mature sequence, was conservative among different species. MiR-27b highly expressed in sheep skeletal muscle than other tissues. In skeletal muscle of Suffolk and Bashbay sheep, miR-27b was upregulated during foetal period and downregulated during postnatal period significantly (P<0.01), but it still kept a relatively higher expression level in skeletal muscle of postnatal Suffolk sheep than Bashbay. There is a potential target site of miR-27b on 3$\text{'}$-UTR of sheep myostatin (MSTN) mRNA, and the double luciferase reporter assay proved that miR-27b could successfully bind on this site. When sheep satellite cells were in the proliferation status, miR-27b was upregulated and MSTN was downregulated significantly (P<0.01). When miR-27b mimics was transfected into sheep satellite cells, the cell proliferation was promoted and the protein level of MSTN was significantly downregulated (P<0.01). Moreover, miR-27b regulated its target gene MSTN by translation repression at an early step, and followed by inducing mRNA degradation in sheep satellite cells. Based on these results, we confirm that miR-27b could promote sheep skeletal muscle satellite cell proliferation by targeting MSTN and suppressing its expression.

4 illus, 2 tables, 37 ref

030243   DING G, ZHANG D, XUE F, GAO J, YEH K-W (Yancheng Institute of Technology, China, Email: dorothy2001@sina.com) : Molecular variation and population structure in endangered Limonium bicolor: genetic diversity of microsatellite markers and amplified fragment length polymorphism analysis. J Genet 2018, 97(5), 1097–105.

Knowledge and analysis of the genetic structure of an endangered species is important for its conservation and evolutionary process. Simple sequence repeats (SSRs) and amplified fragment length polymorphisms (AFLPs) were used in evaluation of the genetic diversity and population differentiation in Limonium bicolor(Plumbaginaceae), an endangered herb with high medicinal and horticulture value. A total of 117 alleles were detected with an average 5.85 alleles per locus using SSR and 222 bands from AFLP were amplified in six populations. It was found that L. bicolor was characterized by high levels of genetic polymorphism (100 and 83.78 %), low levels of total genetic diversity (H_t = 0.2824 and 0.2424), and moderate degrees of genetic differentiation among populations (Φ_ST = 0.284 and 0.251). Analysis of molecular variance (AMOVA) revealed that the main variation component existed within populations (71.56 %; 74.93 %) rather than among populations (28.44 %; 25.07 %). Four main clusters were displayed in the UPGMA using TFPGA, which was consistent with the result of principal coordinate analysis (PCA) using NTSYS. Mutations or infrequent gene flow among populations can increase the plant slowly, thus in situ conservation policies should be implemented first for effective and sustainable development. At the same time, ex situ measures, such as those individuals with rare alleles, to maintain the relationships between individuals and populations are also proposed.

3 illus, 5 tables, 51 ref

030288   NOFAL M Y, ALABODI A B, ALENAZI H K, ALENAZI O H, YAHIA A M B, AL-SHEHRI N Q (Biology Dep, Abdulrahman Univ, Riyadh, Saudi Arabia) : Effect of Commiphora molmol (myrrh) on total leukocyte count and histological alterations in mice. Int J Pharm Res Allied Sci 2018, 7(4), 60-6.

Commiphora molmol has been known as ‘‘myrrh’’ and it has been one of the common herbs consumed throughout the world especially in the Arabian countries. To study the effect of Commiphora molmol on the total leukocyte count and histological alterations in liver and spleen of mice. Adult male mice (20 mice) were adapted in the basal diet in this study, then divided to two groups: The first group contained 10 mice and served as the control group, and the second group consisted of 10 mice and served as myrrh treated group in drinking water. The total leukocytes count was done. Liver and spleen paraffin sections were prepared and stained with H&E to determine the effect of treatments on these organs. The experiment took two weeks. All the experimental testes were done twice in time intervals of one week. Myrrh dietary supplementation improved the cellular immune response by increasing the total leukocytes count in myrrh treated animals when compared to the non-treated animals. In the second group which received myrrh, the livers of the mice suspension showed more and/or less normal histological profile when compared to the first group (control group). Moreover, the microscopic investigation of the spleen of myrrh-cure mice, illustrated an increased density of lymphatic glands around the arterioles in the white pulp. The results from this study demonstrated the effectiveness of myrrh as signs showing future success of immunological drugs, and further investigations should be done to show the benefits of this plant.

7 illus, 1 table, 13 ref

030347   YOUSUFI M K (Microbiology Dep, Jawaharlal Nehru Coll, Bhopal - 462 002, Email: mkhanyousufi@gmail.com) : Analysis of patent laws with reference to microorganisms. Int J Pharm Biol Sci 2018, 8(4), 744-9.

The term microorganisms mean those organisms that are microscopic in nature or that cannot be seen through naked eyes, they consist of both single cell as well as cell clusters [1-2]. The microorganisms are microbiological entities, cellular or non-cellular, capable of replication and transferring genetic material. In the modern era microorganisms are being used to produce variety of biological products like vaccines, antibiotics, proteins, enzymes, beer, wines etc [3]. Various industries like the pharmaceutical, bakery, dairy and the breweries use specific strains of microorganisms for producing specific products [4]. Development of new strains of microorganisms to produce desired products has triggered competition among these industries to patent their respective strains of microorganisms. This has led to develop various patent laws throughout the world for patenting of microorganisms. The present study creeps in in-depth analysis of patenting of microorganisms.

36 ref

030237   DAS S, BORA A, GOGOI H K, MANDERIA S, BAILUNG B, RAJKONWAR J, DWIVEDI S K (Defence Research Laboratory, Tezpur - 784 001, Assam, Email: ajitabhbora@gmail.com) : Effect of growth phase in enhancing PUFA accumulation of few oleaginous freshwater chlorophyceae species. Int J Pharm Biol Sci 2018, 8(4), 642-53.

Freshwater green microalgae are potential sources of vegetarian PUFAs. The fresh water bodies harbor diverse forms of microalgae, which have hitherto remained untapped. With an aim to tap these resources, few green microalgae were evaluated and compared with the different growth phase for their PUFA composition increment with a special interest in omega-3 and omega-6 fatty acid. PUFA content of seven algal species was analyzed by culturing in BG11 medium rich in nitrogen source and grow under unique growth condition established in the laboratory (unpublished). The test species accumulated maximum lipid content during stationary phase when grown in batch mode. GC-MS analysis of lipid extracted during log phase revealed presence of only Linoleic acid, while during stationary phase, other PUFAs were detected such as Docosahexaenoic acid (C22:6) and Stearidonic acid (C18:4) in Chlorococcum humicola, Arachidonic acid (C20:4) in Ankistrodesmus falcatus, Eicosadienoic acid (C20:2) in Desmodesmus opliensis, Hexadecatrienoic acid (C16:3) in Chloralla ellipsoidea and Hexadecatetraenoic acid (C16:4) in Coelastrum microporum. This study thus established a novel growth condition and particular growth phase in which PUFA accumulation enhanced in test fresh water microalgae.

3 illus, 6 tables, 43 ref

030273   MAHAKAL M, KUTUMBALE A, MEHTA D, MEHTA B K (Vikram Univ, Ujjain - 456 010, Email: meenalujjain18@gmail.com) : Green synthesis of ZNO nanoparticles from the methanolic extract of Embelia ribes and their characterization by UV-visible, FTIR, XRD and SEM analysis. Int J Pharm Biol Sci 2018, 8(4), 402-9.

Green synthesis of nanomaterials has become a prominent branch of nanotechnology. In the current study we have reported a cost effective and ecofriendly process for synthesis of Zinc oxide nanoparticles from E. ribes seeds extract. The resultant nanoparticles were characterized using UV–visible spectroscopy, Fourier Transform Infrared Spectroscopy, X-ray diffraction, and Scanning Electron Microscopy. UV-Vis analysis of these microwave assisted ZnO NPs have shown change in color with the time interval of microwave irradiation, which became stable after 8 min of microwave irradiation, hence we can conclude that the reaction was complete. FTIR data reveals the presence of biomolecules which could have reduced the Zn ions to ZnO as well capping and stabilizing agent. Resultant ZnO NPs were found hexagonal in structure, and of dimension 57.57 nm. SEM results found were in accordance with the XRD data obtained from ZnO NPs. Thus, zinc oxide nanoparticles with well defined dimensions could be synthesized from the methanol extract of E. ribes seeds.

6 illus, 2 tables, 22 ref

030325   SUBASANKARI K, THANAPPAN V, ANANTHARAMAN P (Annamalai Univ, Parangipettai, Email: ksshankari.suba@gmail.com) : A comparative study on vitamin B12 and co-culture system promotes the growth of microalgae Nephroselmis astigmatica. Int J Pharm Biol Sci 2018, 8(4), 315-9.

Microalgae frequently grow in marine environment with the help of symbiotic microbes. Here our study explains in detail with the reciprocity between the microalgae Nephroselmis astigmatica and bacteria Halomaonas meridian in habitat with high saline environment. In this study Co- culture system reveals the possibility to know the bacterial metabolites and their growth promotion in algal cells. However algal cells grown in vitamin solution at various concentrations of 5 μg, 7.5 μg, 10 μg reveals the requirement of vitamins for their growth. According to that the co – culture experiment at different incubation period of 12 hr, 24 hr, 48 hr, and 96 hr explains the effect of bacterial growth promoting compounds from late log phase. Hence results of our study show 7.5 μg of cyanocobalamine and 48 hr of Halomonas meridian improves the algal growth beneficially.

2 illus, 28 ref

030265   KAPUR M K, DAS P, KUMAR P, KUMAR M, SOLANKI R (Delhi Univ, New Delhi -110 019, Email: monishaandc@gmail.com) : Molecular characterization of chitinolytic strains isolated from terrestrial habitats. Int J Pharm Biol Sci 2018, 8(4), 239-53.

Three actinomycete strains producing extracellular chitinase, designated as colony 130, 194 and 184 were isolated from soil samples collected from NTPC chemical plant, (Faridabad) Haryana, landfill site (Sarai Kale Khan) Delhi and sugar plant Mandi Dhanaura, U.P respectively. During primary screening diameter of zones of hydrolysis were in the range of 8 to 16 mm as reported in our previous paper Das et al., 2017. Isolates 130, 194, 184 and Streptomyces albidoflavus (NRRLB 16746) as positive control were subsequently subjected to secondary level of screening and purification. Activity of enzyme in crude extracts was in the range of 7.16 -14.12 IU/ml and in partially purified samples in the range 12.1 to 23.10 IU/ml as reported in our earlier studies [14]. Comparison and analysis of the 16S rRNA gene sequence revealed that colonies 130, 194 and 184 belonged to the genus Streptomyces. Colony 130 (Accession no: KY357309) had 97.23% sequence similarity with Streptomyces calvus ISP 5010T (AY999780), colony 194 (Accession no: KJ934595) had 96.20% similarity with Streptomyces albidoflavus DSM 40455T (Z76676) and colony 184 (KY357314) showed 98.8 % similarity with Streptomyces enissocaesilis NRRL B-16365T (DQ026641). Random amplified polymorphic DNA (RAPD) was assayed to determine the genetic diversity of colonies 130 and 194 with reference strain, Streptomyces albidoflavus. Colony 130 showed around 80-90 % interspecific variation, the colony has different genetic origin in comparison to the reference strain. On the other hand, colonies 194 and 184 showed around 50-60 % and 20 % variation respectively and hence have almost same genetic origin as S. albidoflavus.

6 illus, 2 tables, 45 ref

030275   MANIKANDAN P, GNANASEKARAN A, SENTHILKUMAR P K (Microbiology Dep, Annamalai Univ, Chidambaram – 608 002, Tamil Nadu, Email: drpks1980@gmail.com) : Screening and characterization of protease producing halophilic bacteria from saltpan area Vedaranyam, Tamil nadu. Int J Pharm Biol Sci 2018, 8(4), 160-8.

This study was focused on screening and characterization of protease producing bacterial strain from saltpan located in Vedaranyam. The isolated potential bacterial strain was identified as Bacillus firmus by various staining, biochemical tests includes 16S rRNA sequencing method. The effects of various environmental factors on the production of protease were studied. The organism showed the significant growth and enzyme production at pH 9.0 and temperature range at 37ºC. The protease enzyme was analyzed and purified by using ammonium sulphate precipitation method and membrane dialysis. The protease enzyme liberated from these strains showed better stability and activity in pH 8.0 and at temperature 40˚C suggesting that enzyme used in antibacterial activity and pharmaceutical applications.

5 illus, 2 tables, 38 ref

030241   DEEPA V S, ANANTHAN G (Annamalai Univ, Parangipettai – 608 502, Tamil Nadu, Email: casananthan@yahoo.co.in) : Anticancer property of simple ascidian Phallusia nigra Savigny, 1816 against A-549 cancer cell. Int J Pharm Biol Sci 2018, 8(4), 136-48.

Lung cancer is one of the most prevalent disease affects the human beings. To assess the anticancer activity of the ethyl acetate, ethanol, methanol, chloroform and aqueous extract of the simple ascidian Phallusia nigra against human lung cancer cell lines (A-549). The extract administration at 100 μl, 50 μl, 25 μl, 12.5μl, 6.25 μl in 100 μl of 5% MEM. 76% of cytotoxicity was observed in cells cultivated at 100 μl concentration. There was a dose dependent increase in number, inhibition of cancer cells and decrease in size of tumor volume. A high reduction in tumor was noted. In vivo studies indicate the presence of bioactive compounds in Phallusia nigra with anticancer property against A-549 cells.

5 illus, 5 tables, 20 ref

030314   SARAVANAN S, SIVAKAMI R (Zoology Dep, Arignar Anna Govt. Arts Coll, Musiri – 621 211, Email: drsiva17@gmail.com) : A seasonal analysis of microzooplankton in the backwaters of Kottaipattinam, Tamil nadu, India. Int J Pharm Biol Sci 2018, 8(4), 131-5.

The species composition and diversity of various microzooplankton in the backwaters of Kottaipattinam, Pudukkottai, Tamil Nadu, India were assessed for three seasons of the year. Pre-summer season recorded 23 species while pre-summer, 38 species and summer 24 species. Thus, highest diversity was recorded in the pre-summer season. Among the various groups, tintinnids represented 27 species out the total of 38 species and dominated all the three seasons.

2 tables, 20 ref

030315   SARAVANAN S, SIVAKAMI R, PREM KISHORE G (Zoology Dep, Arignar Anna Govt. Arts Coll, Musiri – 621 211, Email: drsiva17@gmail.com) : Actinomycetes diversity in five fresh water systems of Pudukkottai, Tamil nadu and their antimicrobial activity. Int J Pharm Biol Sci 2018, 8(4), 126-30.

Actinomycetes are excellent elaborators of biotechnological products such as antibiotics, industrial enzymes and other bioactive compounds. Emergence of multiple drug resistant pathogens implies the need to search for new and novel antimicrobials and the discovery of new molecules from actinomycetes has marked an epoch in antibiotic research and subsequent developments in antibiotic chemotherapy. Sediment samples were collected from five different ponds of Kottaipattinam region, Pudukkottai District situated near the Bay of Bengal. Actinomycetes strains were isolated from the air-dried sediment samples by applying serial dilution and spread plate technique on three different media such as Glycerol-Arginine Agar, Starch-Casein Agar, and Starch-Nitrate Agar. Different media were used in order to compare the recovery on different media as well as to obtain maximum diversity of actinomycetes strains. The test organisms used were bacteria such as Escherichia coli, Klebsiella pneumoniae, Salmonella typhi, Bacillus cereus, Vibrio cholerae, Salmonella paratyphi and Staphylococcus epidermidis. The test fungus/yeast used were Aspergillus niger and Candida albicans. The test organisms were collected from Government Hospital, Tiruchirappalli, Tamil Nadu. In the present work, the antibacterial activity of actinomycetes strains were checked by using three different media (such as Glycerol Argine Agar (GAA), Starch- Casein Agar (SCA) and Starch Nitrate Agar (SNA) and three methods (like cross streak method, spectra-plate method and well diffusion method). However, the strains showed antibacterial activity only by well diffusion method on Glycerol-Yeast Extract Agar for bacteria and Potato Dextrose Agar for fungi. Thus, it is concluded that the substances repressing these strains were accumulated only in the mycelium and not in the medium.

3 tables, 17 ref

030320   SIVAKAMI R, SARAVANAN S, PREM KISHORE G (Zoology Dep, Arignar Anna Govt. Arts Coll, Musiri – 621 211, Email: drsiva17@gmail.com) : Bacterial biosurfactants and their role in heavy metal degradation. Int J Pharm Biol Sci 2018, 8(4), 113-9.

Biosurfactants are produced by certain specialized organisms including bacteria fungi and yeast. These microbes produce biosurfactant either secreted or attached to parts of the cell membrane predominantly during growth phase. Microorganisms producing biosurfactants help to amplify the bioavailability of hydrocarbons by enhancing the contact between the pollutants and microorganisms in the presence of the biosurfactant which helps in the accelerated bioremediation of hydrocarbon contaminated sites. Hence the present study was carried out to know the biosurfact producing potential of two bacterial species (Bacillus subtilis and Pseudomonas aeruginosa) and their effect on degrading two metals (zinc and iron). Results show that both the species helped in the reduction of heavy metals zinc and iron. However, among the two B. subtilis appeared to be a better candidate as it recorded a higher conversion rate.

3 tables, 49 ref

030313   SARAVANAN S, SIVAKAMI R (Arignar Anna Govt. Arts Coll, Musiri - 621 211, Email: drsiva17@gmail.com) : Identification of Vibrio species occurring in food fishes and studies on their antibiotic resistances. Int J Pharm Biol Sci 2018, 8(4), 103-6.

Of the Crustaceans suitable for aquaculture, shrimp plays an important role. To increase production, aquaculture have resorted to high stocking densities leading to diseases resulting in heavy losses to aquaculture industry. Among the various pathogens, Vibrio has been found to be one of the most studied and diverse genus of microorganisms found in the aquatic systems. Hence, the present study was attempted to identify the different species of Vibrio that occurs in fishes and shrimps grown in this part of the country during the different seasons. A total of 11 species of Vibrio could be isolated of which four species dominated during the rainy season and two species each during the pre-summer and summer season. The differences in the distribution and occurrence of these species is attributed to the changes in the physicochemical and climatic conditions during each season. Studies on antibiotic resistance reveals that among the various substances tested, herbal extract appeared to be the most effective.

2 tables, 13 ref

030278   MATHARASI A, KUMAR R D, PRABAKARAN G, KUMAR P S (Annamalai Univ, Tamil Nadu – 608 502, Email: sampathcas@gmail.com) : Phytochemical screening and antimicrobial activity of marine microalgae Tetraselmis sp. Int J Pharm Biol Sci 2018, 8(4), 85-90.

The aim of the present study was to screen the bioactive chemicals present in Tetraselmis sp. extracted with various solvents such as Methanol, Hexane, Diethyl ether. Antimicrobial potential of Tetraselmis sp. extracts was determined by Agar disc diffusion method against human pathogens of both Gram positive (E.coli, Proteus vulgaris) and Gram negative bacteria (Pseudomonas aeruginosa, Bacillus subtilis, Staphlococcus aureus) and three fungal pathogens such as Aspergillus fumigatus, Aspergillus niger, Candida albicans, were taken and studied against the standard streptomycin and chloramphenicol. All the extractions were subjected to evaluated for the presence of the active chemical compounds and alkaloids, tannins, saponins, flavonoids, and steroids are the major phytochemicals present in the extractions. Maximum inhibitory effect was noticed in Methanolic extracts against E.coli (16 mm) and S. aureus (15 mm) whereas fungal pathogens Aspergillus niger (7 mm). Negative effect was observed against Proteus vulgaris, Candida albicans and Aspergillus fumigatus in all the extracts of Tetraselmis sp. The antibacterial effectiveness might be due to presence of the active compounds present in the Methanolic extract and having promising pharmaceutical applications.

1 illus, 4 tables, 30 ref

030303   RAMYA H, VEENA R K, JANARDHANAN K K (Amala Cancer Research Centre, Thrissur, Kerala, Email: kkjanardhanan@yahoo.com) : Anti-tumour activity of exo-polysaccharides produced by an isolate of Ganoderma lucidum occurring in South India. Int J Pharm Biol Sci 2018, 8(4), 20-4.

Ganoderma lucidum is a popular medicinal mushroom worldwide because of its fascinating therapeutic properties. Investigations carried out in our laboratory have demonstrated the antitumor activity of the extracts, polysaccharides and the terpenoids isolated from fruiting bodies of G. lucidum occurring in south india. In this study, we examined the antitumor activity of the exopolysaccharides produced by a G. lucidum isolate in submerged culture. The exopolysaccharides were isolated from the culture broth by repeated ethanol precipitation, treatment with Sevag’s reagent for the removal of protein, dialysis and lyophilisation. The antitumour activity was tested on Swiss albino mice implanted with Dalton’s lymphoma ascites (DLA) cell line. The exopolysaccharides at doses of 5, 10 and 25 mg/kg body weight achieved 80, 84 and 86 % tumor growth inhibition respectively. The standard reference drug, cyclophosphamide at a dose 25 mg/kg body weight showed 94 % tumor inhibition. The results thus showed that exopolysaccharides produced by the G. lucidum isolate possessed excellent antitumor activity. The finding suggests the potential therapeutic use of exopolysaccharides produced by G. lucidum.

2 illus, 2 tables, 14 ref

030232   CHATURVEDI R, GAUTAM S K, KAUR S, AHMED F, KAORE N M (Microbiology Dep, Raipur Institute of Medical Sciences, Raipur - 492 101, Email: drshankgautam@gmail.com) : Assessment of microbial contamination of toothbrushes and factors influencing the same in medical students. J Evolution Med Dent Sci 2018, 7(52), 5565-9.

Oral health is an integral part of good general health and toothbrushes are the most common and vital component of daily maintenance of oral hygiene used for prevention of oral diseases. Unfortunately, public awareness is lacking on the unsanitary conditions in which the brushes are placed like bathrooms, attached toilets in bathrooms which makes it prone for contamination with variety of microbes. To evaluate the contamination of the tooth brushes used and stored in different environmental conditions by medical students for various microbes and prevention of the same by using 0.2 % Chlorhexidine gluconate. This cross sectional prospective analytical study was carried out in the Department of Microbiology attached to Medical College during a time period of 2 months from 1^st July 2018 to 31^st August 2018 after due approval from Institutional Ethics Committee (IEC) on 45 Second MBBS medical students consenting for the proposed study. They were grouped into 3 groups based on the storage condition of the tooth brush viz. 1- Dry place outside bathroom, 2- Inside bathroom without attached toilet & 3- Inside bathroom with attached toilet. After initial assessment of the commensal flora they are possessing by a new brush, they were given two sets of tooth brushes, one of red colour and other of blue colour. The red brush was the morning brush and was stored after rinsing with tap water as it is. The blue brush was the sleep time brush and was rinsed and dipped in 0.2 % Chlorhexidine gluconate solution provided to them for 15 minutes, dried and stored. The microbial contamination in red brush stored in various environmental condition as well as bacterial load was noted. The blue brush was assessed for efficacy of 0.2 % Chlorhexidine gluconate used in different environmental conditions in which the brushes were stored by different group members. Data was maintained in Microsoft Office Excel and analysed by tests of proportion and significance. Of the 45 volunteered medical students in the present study between the age group of 19 to 25 years, gender distribution was 33 % males against 67 % females with a Male to Female ratio of 1:2. Of the 45 participants assessed 11/45 (24.44 %) grew Streptococci, 8/45 (17.77 %) Staphylococcus, 3/45(6.66 %) Gram positive bacilli, 7/45 (15.55 %) Pseudomonas and 01/45 (2.22 %) of E. coli. The overall efficacy of 0.2 % Chlorhexidine gluconate was assessed in terms of reduction of the pathogenic strains and found to be most effective in Group 2 with p-value of <0.001 followed by Group 3 with p-value of <0.05 indicating significant reduction in contamination by pathogenic bacteria whereas no significant efficacy was found in Group 1. The results of the present study established that tooth brushes are prone to bacterial contamination in dry as well as wet environments and can be a cause of potentially dangerous infections and spread of it, thus requiring adequate care to prevent the same. The appropriate methods of preservation of tooth brushes following oral hygiene practice needs attention & use of 0.2 % Chlorhexidine gluconate can definitely reduce the chances of bacterial contamination of toothbrushes significantly especially in wet conditions of bathroom storage.

2 tables, 18 ref

030228   BHATTACHARYYA I, BANERJEE D (Microbiology Dep, CNMC, Kolkata - 700 103, Email: debarati.ban@gmail.com) : The terrorist still lurks in the hospital- Prevalence of MRSA. J Evolution Med Dent Sci 2018, 7(52), 5509-12.

MRSA poses a very serious problem producing difficult to treat infections in the community as well as in health facilities. Out of these, Health Care associated MRSA infections have the additional problem of being multidrug resistant. This fact led us to conduct this study in a tertiary care hospital in Kolkata. Samples of pus were collected from all the wards of the hospital. Samples were received within 30 minutes of their collection. The usual protocol of Gram stain, culture, biochemical tests and antibiotic sensitivity was followed. Phenotypic method using 30 µg cefoxitin disc was followed for identifying MRSA according to CLSI guidelines. The results were tabulated, and prevalence of MRSA was calculated. Out of 102 Staphylococcus aureus samples 35.3 % were found to be methicillin resistant. This prevalence of MRSA is found to be at par with a study done by Sangram S Patel et al. in 2015. Thus, the results of the present study show a high endemicity of MRSA in our hospital in Kolkata. Though our results show 50 % of the MRSA were MDR which could be due to selective antibiotic pressure. But fortunately, none of the isolates were XDR or PDR. This poses a serious problem for drug therapy because the treatment options become limited to potentially toxic antimicrobials like vancomycin, teicoplanin, streptogramins, linezolid and ceftaroline leading to increased mortality and morbidity.

3 illus, 18 ref

030318   SINGH N H, SINGH R M, SINGH H L, DEVI Y B (Microbiology Dep, Jawaharlal Nehru Institute of Medical Sciences, Imphal - 795 005, Email: rkmksingh@gmail.com) : Seroprevalence of Hepatitis B virus infection among patients attending a tertiary care centre in north-east India. J Evolution Med Dent Sci 2018, 7(52), 5506-8.

Hepatitis B infection is a major global health problem. The seroprevalence of Hepatitis B virus infection among general population attending at a tertiary care hospital is useful in assessing the magnitude of HBV infection and helps in devising preventive measures. The co-infection of Hepatitis B or Hepatitis C virus with HIV accelerates disease progression and may complicate the management of patients infected with HIV. This study was designed to determine hepatitis B virus infection among patients attending tertiary care centre in North-East India. A total of 11399 serum samples, collected from patients attending outpatient or inpatient departments and ART centre, were screened for HBsAg using 3rd generation ELISA (Qualisa HBsAg, Qualpro Diagnostics, Goa, India) in this retrospective study. Seroprevalence of HBsAg was 1.23 %. Seropositivity was seen more in males (2.2 %) as compared to females (0.89 %). Among males, highest occurrence was found in the age group of 21-30 (3.29 %) years followed by 31-40 (2.94 %) years and 41-50 (2.37 %) years. Among females, highest prevalence was observed in age group of 51-60 (1.39 %) years followed by 61-70 (1.15 %) years and 41-50 (1.13 %) years. Present study showed that the overall rate of HBsAg positivity in our center was 1.23 %. Males and younger age group were found to be more affected probably due to horizontal transmission.

3 tables, 17 ref

030266   KAUR M, SIDHU S K, MALHOTRA S, MALHOTRA S K, DEVI P (Microbiology Dep, Government Medical Coll, Amritsar, Punjab, Email: shail78@hotmail.com) : A study of prevalence of dermatophytes in a tertiary care hospital of northern India. J Evolution Med Dent Sci 2018, 7(48), 5203-6.

Superficial fungal infections have become a significant health problem affecting all age groups. Skin infections due to dermatophytes are caused by a group of closely related keratophilic fungi, capable of invading the keratinized tissues of skin. This study was undertaken to determine the prevalence of dermatophytosis, and to identify the dermatophytes from skin samples of clinically suspected cases in a tertiary care hospital. A total of 197 skin samples from suspected cases of dermatophytosis were received and processed in the department of microbiology. All the samples were subjected to microscopic examination and culture by standard techniques. Dermatophytes were identified by studying macroscopic and microscopic characteristics of the isolates. Among the clinically suspected cases (197) of dermatophytosis, most common age group was 21-40 years (52.3 %). Fungal elements were detected in 75 (38.08 %) cases either by direct microscopy or by culture. Trichophyton species were the most common dermatophytic species isolated (87.1 %) followed by Microsporum species (12.9 %). Accurate assessment of the prevalence of dermatophytic infections, early laboratory diagnosis and timely treatment can limit the spread of these infections.

7 illus, 1 table, 21 ref

030242   DESHMUKH V, RAJEBHOSALE R, MENCH K, JOSHI M (General Surgery Dep, Rajashree Chatrapati Shahu Maharaj Government Medical Coll, Kolhapur, Maharashtra, Email: ramraje007@gmail.com) : The study of incidence of biliary infection in gall stone disease in a hospital. J Evolution Med Dent Sci 2018, 7(47), 5107-12.

Biliary calculus disease is one of the most common disorders affecting the gastrointestinal tract and is an important cause of morbidity. In about half of the cases, bacteria can be cultured from the gallbladder bile. Infective factor seems to be a major cause of formation of gallstones. Moynihan’s aphorism that “gall stone is a tomb stone erected in the memory of the organism with in it” is true today.1 Evidence in favour of infection includes isolation of E. coli, bacterium typhosum, streptococcus from the gallbladder bile and from the centre of the gallstones.2 Knowing the bacteriological aetiology will help in prevention as well as better treatment of gallstone disease in the future. This is descriptive study which includes cases of cholelithiasis, age between 21 to 80 years, who underwent cholecystectomy from June 2014 to June 2016 at CPR Hospital attached to RCSM Government Medical College Kolhapur . Cases of acute cholecystitis, acute acalculous cholecystitis, empyema, mucocele were excluded from the study. The objectives of this study were 1. to determine frequency of infection in cholelithiasis, 2. to find common infecting organisms and 3. To determine their antibiotic sensitivity. Sample size was calculated as 100 using Sample Size formula Sample size =Z 2 * (p) * (1-p)/c 2. Relevant laboratory investigations like CBC, BSL, LFT with liver enzymes were done. Relevant radiographic investigations like USG were done. Patients were operated either by open or laparoscopic technique. Antimicrobial prophylaxis with 1 gm Ceftriaxone preoperatively and patients who had diabetes mellitus or spillage of bile or stone into the peritoneal cavity continued receiving 1 gm Ceftriaxone every 12 hours postoperatively. Bile and gallstones collected by sterile technique were then put into sterile bottles and transferred to laboratory where they were crushed and cultured to identify organisms and their sensitivity. Statistical analysis was done by Chi square test and Fisher’s exact tests. All patients had cholelithiasis and chronic cholecystitis. Bacteria were isolated in 36 samples (36 %). Frequency of isolated bacteria in females and males had no significant difference (P > 0.05). Monomicrobial and polymicrobial infection were observed in 34 (94 %) and 2 (6 %) respectively. E. coli was the most common isolate (12; 33 %). Enterobacter was the second one (8; 22 %). The antibiotics to which organisms were sensitive were Cefepime, Ceftriaxone, Imipenem, Amikacin, Gentamycin and Ciprofloxacin.1. Most common organism detected in bile culture was E. Coli followed by enterococcus and anaerobic bacteria. 2. Most effective antibiotic in bile and stone culture positive cases were Cefepime, Ceftriaxone, Imipenem, Amikacin, Gentamycin and Ciprofloxacin. 3. We recommend starting antibiotics selectively, if supported by the clinical conditions or co-morbidities like diabetes or immune-compromised states and culture positive in SSI wound infections according to its culture and sensitivity, in case of cholecystitis and cholelithiasis.

3 illus, 3 tables, 14 ref

030257   JAGDISH P, ASHOKA R K (Anatomy Dep, K. D. Medical Coll Hospital and Research Center, Mathura, Uttar Pradesh, Email: prerna.jagdish27@gmail.com) : Morphometric analysis of the human spleen. J Evolution Med Dent Sci 2018, 7(46), 5011-4.

With respect to its size, shape, weight, notches & fissures, spleen is a highly variable organ. It is otherwise very difficult to analyse whether the spleen is normal or abnormal on the basis of its shape; therefore this study has been undertaken to find out percentage of various shapes and other morphometrical parameters of spleen. Accurate knowledge of splenic morphology is essential prior to performing any surgical procedure or making any radiological interpretation with regard to spleen as it reveals many variations in shape, size, notches & fissures. The present study is a descriptive study conducted in the Department of Anatomy, K. D. Medical College, Mathura, U.P. 60 adult human cadaveric spleens were included. Sexual dimorphism was excluded from the research. The spleens were dissected from the abdominal cavity and a morphometric analysis was done with the help of digital sliding calipers. Data obtained was compared with data available from previous studies after its tabulation and statistical analysis. 60 human spleens were morphologically analysed in detail. Wedge-shaped spleens were most commonly noted. Mean length was calculated to be about 10.5 cm. Mean breadth was calculated to be about 7.5 cm. Mean width was calculated to be about 4.25 cm. Studies on the morphometry of spleen will be of interest not only from academic point of view but also for operating surgeons and interventional radiologists. Detailed knowledge on spleen is important to avoid and prevent any complications and to obtain a good operative, as well as diagnostic intervention.

5 tables, 23 ref

030225   ANSAR S, DASTAGIR A (Microbiology Dep, Coimbatore Medical Coll, Coimbatore, Tamilnadu, Email: drdayesha@gmail.com) : Bacteriological profile of diabetic foot ulcers. J Evolution Med Dent Sci 2018, 7(46), 4997-5002.

Diabetes mellitus is a progressive disease, diabetic foot is one of the major complications of diabetes mellitus and eventually leads to development of gangrene and lower extremity amputation. This study has been carried out to detect the antibiotic sensitivity pattern of the isolates in diabetic foot ulcer patients. This is a descriptive cross-sectional study. Pus and wound swabs were collected from around 100 diabetic patients with foot ulcer who attended the Surgery Out-Patient Department of Coimbatore Medical College Hospital. The samples received in the Department of Microbiology were processed for aerobic culture and antibiotic sensitivity testing during the study period. Blood samples were collected to analyse the HbA1c levels. Of the 100 cases studied, most of the patients belonged to the 5th and 6th decades of life (37 %) and (28 %) respectively. Males were more affected compared to females with a ratio of 2.3:1. Maximum number of patients with Diabetic Foot Ulcers were seen in Wagner’s Grade II (40 nos.), followed by 38 DFI patients in Wagner’s Grade III. Among Gram positive aerobes, Staphylococcus aureus was the predominant isolate (18.4 %). Among Gram negative aerobes, Proteus spp. was the most common isolate (23.2 %) followed by E.coli 16.8 % and Pseudomonas 16 %. Acinetobacter species was the least common isolate (2.4 %). While staphylococcus aureus was the most common among gram positive cocci, proteus was the most common isolate among the gramnegative pathogens. Staphylococci and Proteus were the two most common isolates detected in diabetic foot infections. There was significant association between DFI and higher HbA1c levels. But no correlation was found between HbA1c levels and the polymicrobial nature of infection in DFI.

6 tables, 22 ref

030338   VIJAYA, NAGAR S N (Microbiology Dep, Srinivas Institute of Medical Sciences and Research Centre, Mangalore, Karnataka, Email: sweetsnayak@yahoo.com) : Rising trend of ESBL production in uropathogenic E. coli causing worry? - A study in a tertiary care hospital. J Evolution Med Dent Sci 2018, 7(46), 4938-42.

Urinary Tract Infection (UTI) caused by Escherichia coli (E. coli) has become a significant global public health problem. The resistance to β-lactam antibiotics in E. coli further adds to the problem. The pattern of drug resistance among E. coli undergoes considerable variation and consequently the treatment of Urinary Tract Infection (UTI) requires constant updating of the antibiotic sensitivity profile. The study was thus undertaken to know resistance pattern of E. coli isolates with special reference to production of extended spectrum β-lactamases (ESBLs) in urinary samples. This is a three-year (2015 - 2017) retrospective descriptive study undertaken in Srinivas Institute of Medical Sciences and Research Centre, Mukka, Mangalore. Standard culture techniques for urine samples were followed. Antibiotic sensitivity test was done by Kirby-Bauer disc diffusion method and interpretation was done following Clinical and Laboratory Standards Institute (CLSI) guidelines. ESBL detection was done by NCCLS phenotypic confirmatory combination disc diffusion method using ceftazidime (30 μg) alone and ceftazidime + clavulanic acid (30 μg/ 10 μg). E.coli is the most frequent isolate found in UTI (65.4 % of total isolates) throughout three years. The overall resistance pattern of E.coli exhibited highest resistance against ampicillin (95 %) and amoxyclav (83.3 %) with rising trend in resistance to Cephalosporins and Quinolones in three successive years. The rising trend in ESBL production rate was observed from 27.3 % (2015) to 39.4 % (2017) with least resistance against meropenem (9.9 %), piperacillin/ tazobactam (10 %), nitrofurantoin (11.4 %) and amikacin (12.1 %). It was also observed that > 50 % of ESBL producing E. coli isolates exhibited resistance against cotrimoxazole and Quinolones. The incidence of the ESBL producing E. coli has been steadily increasing over the past few years. The knowledge on the resistance pattern in a geographical area will help in guiding the appropriate and the judicious use of antibiotics.

4 tables, 25 ref

030337   VERMA S, THORAT S S, BISURE K (Microbiology Dep, Group of TB Hospital, Mumbai, Maharashtra, Email: drvermasukanya@gmail.com) : Prevalence of carbapenem resistant enterobacteriaceae- A study in a tertiary care hospital in Mumbai. J Evolution Med Dent Sci 2018, 7(45), 4909-12.

The global incidence of carbapenem resistant organisms is on the rise and can be attributed to indiscriminate use of carbapenem to tackle the Extended Spectrum Beta Lactamase (ESBL) producing organisms. Carbapenem resistance is associated with resistance to other group of antibiotics. The spread of resistance from one bug to another is not uncommon. The organism persists in the hospital environment and is identified as cause of hospital-acquired infections. This is not only a major concern in the healthcare setting but is also increasingly being recovered from community settings also. Limited availability of drugs to treat these infections is the biggest concern. Identifying carbapenem resistant organisms and implementing measures to prevent the spread is need of the hour. The aim of this study is to determine the prevalence and susceptibility pattern of Carbapenem resistant Enterobacteriaceae. A descriptive study was carried out in the Department of Microbiology of a tertiary care teaching institute of Mumbai. All Enterobacteriaceae isolates recovered during the six months study period of January 2017 to June 2017 were included in the study. The isolates were identified using standard conventional biochemical methods. Antimicrobial susceptibility was performed using the Kirby-Bauer disc diffusion technique and interpreted as per CLSI 2016 standards. A total of 442 isolates of Enterobacteriaceae were recovered during the study period. The different strains isolated included Escherichia coli (218), Klebsiella pneumoniae (161), Proteus spp. (42), Salmonella (9), Citrobacter spp. (1) and Enterobacter spp. (1); 107 out of 442 (24.2 %) were Carbapenem resistant 18.8 % of E. coli, 37.9 % of Klebsiella pneumoniae. Maximum number of CRE were recovered from Medicine Department (34.1 %). The least number of CRE isolates were recovered from ENT (5.8 %). The prevalence of Carbapenem resistant isolates is high in our settings. Strict adherence to infection control practices and stringent implementation of antimicrobial stewardship is essential to curb the rate of Carbapenem resistant isolates.

4 tables, 21 ref

030224   ANSAR S, DASTAGIR A (Microbiology Dep, Perundurai Medical Coll, Erode, Tamilnadu, Email: shajahan47878@gmail.com) : Bacteriological profile of diabetic foot ulcers with reference to HbA1c levels. J Evolution Med Dent Sci 2018, 7(45), 4853-6.

The prevalence of diabetes is increasing in India. Patients with diabetes mellitus may have many serious sequelae. Among them, foot ulcers are most common and may lead to severe complications. The present study was undertaken to determine association between HbA1c levels in relation with diabetic foot infections, bacteriological profile and antibiotic susceptibility pattern. The descriptive study was carried out in the Department of Microbiology, CMC from March 2009 to Sep 2010 to look for the pattern of growth of aerobic organisms in diabetic foot ulcers. Of the 100 DFI cases studied, most of the patients belonged to the 5th and 6th decades of life, (37 %) and (28 %) respectively. Males were more affected compared to females with a ratio of 2.3: 1. Maximum number of patients were seen in Wagner’s Grade II (40 nos.) followed by Wagner’s Grade III (38 nos.). Maximum number of diabetic foot ulcer cases (45) were recorded with HbA1c levels of > 10. The total distribution of monomicrobial and polymicrobial growth was 62 and 28 respectively with the maximum number of growth recorded in HbA1c levels of more than 10. MRSA constitutes 50 % in HbA1c levels of < 8 and 8 to 10 and 55 % in HbA1c levels of > 10. The incidence of diabetic foot lesions strongly correlates with poor glycaemic control, which in itself is best manifested by the levels of glycosylated haemoglobin levels. There was significant association between DFI and higher HbA1c levels.

6 tables, 26 ref

030230   CHAMKHI I, SBABOU L, AURAG J (Mohammed V Univ in Rabat, Rabat 10000, Morocco, Email: chamkhi.imane@gmail.com) : Endophytic fungi isolated from Crocus sativus L. (saffron) as a source of bioactive secondary metabolites. Pharmacogn J 2018, 10(6), 1143-8.

Endophytic fungi are becoming an important source of new natural bioactive products. Many interesting endophytic fungi have been isolated from traditional medicinal plants. Crocus sativus L. (saffron) is one of the most expensive and rarest spices in the world, used as a dye, aroma and for medicinal purposes. This study reports on the molecular characterization of endophytic fungi isolated from roots of saffron plants growing in Taliouine-Morocco, and the examination of the antibacterial and antioxidant activities of secondary metabolites extracted from these endophytes. The fungi were isolated from sterilized saffron root fragments and identified using internal transcribed spacer (ITS) rRNA gene sequence analysis. Three endophytes were fermented and extracted with ethyl acetate (EtOAc) and chloroform (CHCl₃). The antibacterial activity of fungi extracts was evaluated using the Agar Diffusion Method against six strains, Bacillus sp., Stenotrophomonas sp., Pseudomonas putida, Pantoea sp., Luteibacter sp. and Escherichia coli. The antioxidant activity was tested by using the α,α-diphenyl-β-picrylhydrazyl (DPPH) analysis and β-Carotene bleaching test (BCBT) methods. Sixty fungal isolates were recovered and purified from saffron roots. ITS rDNA sequences were 99-100% identical to three different species: Rhizopus oryzae, Aspergillus fumigatiaffinis and Aspergillus niger. Both Rhizopus oryzae extracts had high antibacterial activity against most of the bacteria tested, while Aspergillus niger and Rhizopus oryzae extracts showed an antioxidant capacity using DPPH and BCBT methods, respectively. The fungal endophytes inhabiting saffron roots could be a potential source of natural plant bioactive secondary metabolites especially for antibacterial and antioxidant purposes.

3 illus, 2 tables, 33 ref

030326   SUHARTY N, WAHYUNI F S, DACHRIYANUS (Andalas Univ, West Sumatra, Indonesia, 25163, Email: dachriyanus@ffarmasi.unand. ac.id) : Cytotoxic activity of ethanol extract of arbuscular mycorrhizal fungi induced ginger rhizome on T47D breast cancer cell lines. Pharmacogn J 2018, 10(6), 1133-6.

A study of investigate the cytotoxicity activity of ethanolic extract of ginger (Zingiber officinale Rosc.) induced with arbuscular mycorrhizal fungi (AMF) against T47D cells line breast cancer have been conducted. Cytotoxicity were determined using the “microtetrazolium (MTT) Assay”, by measuring the activity of mitochondrial dehydrogenase in living cells that have ability to convert pale yellow of dissolved MTT to purple formazan product. The extract used at various concentration (0.1, 1.0, 10 and 100 μg / mL. The level of cytotoxic actifity was determined by calculating the inhibitory concentration (IC₅₀) value that was based on the precentage of cell death after 24 h treatment with the extract. The change of cell morphology were observed by using inverted microscope. The statistic results proved that ethanol extract of AMF induced ginger rhizome could barriers T47D breast cancers significantly at concentrations of 10 μg / mL and 100 ug / mL, with IC₅₀ value was 12.5 ± 3.73 μg / mL. centration of 0.1 μg / mL, 1.0 μg / mL, 10 μg / mL and 100 mg / mL. Results of statistical analysis showed that the ethanol extract of ginger rhizome induced AMF at a concentration of 10 μg / mL and 100 μg / mL was able to inhibit the growth of breast cancer cells T47D significantly. The results showed the ethanol extract of AMF induced ginger rhizome was potential as herbal medicine for cancer-related ailments with IC₅₀ value was 12.5 ± 3.73 μg / mL.

2 illus, 2 tables, 11 ref

030331   TEJA K V, RAMESH S , PRIYA V (Conservative Dentistry and Endodontics Dep, Saveetha Univ, Chennai - 600 077, Email: dean.acd@Saveetha.com) : Regulation of matrix metalloproteinase-3 gene expression in inflammation: A molecular study. J Conserv Dent 2018, 21(6), 592-6.

Matrix metalloproteinases (MMPs) play a significant role in the efficient tissue turnover and remodeling. This study focuses on the regulation of the MMPs by the protein kinases at the level of gene expression and their signaling pathways. Lipopolysaccharide‑induced murine macrophage‑like RAW 264.7 cell lines were obtained and maintained in Dulbecco’s modified Eagle’s medium plus 10 % fetal bovine serum under optimal temperatures. Primers used were MMP‑3 forward primer, MMP‑3 reverse primer, and glyceraldehyde‑3‑phosphate dehydrogenase forward primer and glyceraldehyde‑3‑phosphate reverse primer. Total RNA was isolated, the sample was prepared, and electrophoresis was performed. The first strand of cDNA was synthesized and amplification of specific isolated gene using polymerase chain reactor (PCR). The amplified products were then separated on a 1.0 % agarose gel in 1XTBE at 75 V for 3 h. The gel was stained with ethidium bromide, and the amplified product was visualized and photographed on Gel Doc system. Real‑time PCR showed only bands at expected size of 595 bp for internal control amplification of glyceraldehyde‑3‑dehydrogenase gene. Analysis was done with densitometry, and these values are compared with the negative control. Results showed a statistically significant rise in the relative levels of MMP‑3‑mRNA when compared with negative control at 1, 2, and 3 h. This study proved the significantly increased levels of MMP gene at different period, thereby it can be concluded that MMP‑3 levels are higher in inflammatory conditions.

2 illus, 3 tables, 25 ref

030245   DOGAN H H, DUMAN R, TUNCER P (Biology Dep, Selcuk Univ Konya, Turkey, Email: hhuseyindogan@yahoo.com) : Effect of Morchella conica and Fomes fomentarius extracts against the human respiratory syncytial virus in-vitro. Int J Pharm Sci Res 2018, 9(12), 5240-5.

Antiviral properties of methanol and aqueous extracts from Morchella conica and Fomes fomentarius were assessed by colorimetric XTT test against human respiratory syncytial virus (HRSV). HEp-2 cell line was used for the cytotoxic and antiviral effects of the extracts. Non-cytotoxic concentrations of methanol and aqueous extracts of M. conica were tested in 50000 to 195 µg/mL. In contrast, the cytotoxic F. fomentarius methanol and aqueous extracts and Ribavirin were tested at concentrations from 3347-13.1 μg/mL, 4906.5-19.2 μg/mL and 86.60- 0.30 μg/mL, respectively. The results showed that aqueous extract of F. fomentarius had strong anti-HRSV activity (EC₅₀ = 358.7 μg/mL, SI = 27.4) which could be compared with ribavirin (EC₅₀ = 15.6 μg/mL, SI = 11.1) used as a positive control against HRSV, while other extracts (M. conica methanol extract EC₅₀ = 28055 μg/mL, SI = >1.8; F. fomentarius methanol extract EC₅₀ = 3000 μg/mL, SI = 2.2) were found to have weak antiviral activity. As a result, it can be said that aqueous extract of F. fomentarius is worthy of further study as an alternative drugs in order to develop anti RSV.

3 tables, 33 ref

030255   IIKASHA A M N, DUSHIMEMARIYA F, MBEWE M, EIMAN H, MUMBENGEGWI D R (Anatomy Dep, Namibia Univ, Windhoek, Namibia, Email: aiikasha@unam.na) : Antimicrobial and antiplasmodial activity displayed by actinomycetes isolated from soils from Kavango and Hardap regions in Namibia. Int J Pharm Sci Res 2018, 9(12), 5191-6.

Plasmodium falciparum and multidrug resistant bacteria such as Mycobacterium avium, Staphylococcus aureus and Escherichia coli are among the top causes of morbidity and mortality especially in developing countries. These pathogens are responsible for the reported antibiotic resistance incidents globally. Due to the increase in antibiotic resistance worldwide, there is an urgent need for new bioactive compounds. This study aimed at determining the antiplasmodial and antimicrobial activity of secondary metabolites produced by soil actinomycetes. Soil samples were collected from Kavango and Hardap regions and cultured on various media to isolate and identify actinomycetes. Secondary metabolites produced were extracted using different solvents. The antimicrobial activity of different isolated secondary metabolites was determined against M. avium, S. aureus and E. coli. Antiplasmodial activity of four bioactive fractions with potent antibacterial activity was further determined against a chloroquine resistant (D10) P. falciparum strain at various concentrations. Bioactive compounds from two isolates displayed activity against M. avium, whilst another two showed activity against E. coli. None of the secondary metabolites displayed activity against S. aureus. A concentration dependent reduction in parasitaemia for P. falciparum was observed across all four fractions both after 24 and 48 h. These results display a vast potential antibacterial and antiplasmodial activity and should be considered for future drug development

2 illus, 4 tables, 20 ref

030297   PAUL D, PAUL K (Pharmacology Dep, Vivekananda Coll of Pharmacy, Bengaluru - 560 055, Email: deeprajcology81@gmail.com) : Aspergillosis: An overview. Int J Pharm Sci Res 2018, 9(12), 5032-49.

Aspergillus produces both airborne and waterborne infections with deadly consequences depending on the species involved and the immune status of patients. Patients may remain asymptomatic for up to 18 months. The severity of aspergillosis is mainly seen in HSCT recipients with mortality up to 90 %. Though aspergillosis is primarily opportunistic and lung is the primary site of infection yet deviation observed where the primary site was organs other than lungs. The major reasons behind invasion are prolonged (> 3 weeks) high dose corticosteroid therapy, broad-spectrum antibiotics, CMV infection, iron overload, decreased neutrophil count (< 500 cells/mm³), etc. Monoclonal technology in the form of EB-A2 and JF5 antibody can identify fungal antigen much before the appearance of clinical symptoms. Selection of a test and its success to determine fungal invasion depend on associated morbidity as galactomannan assay is more suitable in HSCT recipients rather than solid organ transplant recipients whereas PCR is specific for cancer patients with thrombocytopenia to differentiate between aspergillosis and histoplasmosis. Thorough knowledge about the neutrophil profile is required to interpret the results of CT scan and biopsy. The success of treatment depends on early and correct diagnosis as Aspergillus mimics precancerous conditions and there is considerable overlapping of symptoms between different species of Aspergillus and therapy must be species specific. Mutations in Cyp51A, L98H, M220I, F219C, and G54W are responsible for azole resistance, but the mystery of resistance did not resolve completely as certain fungal isolates show no such mutations but are resistant to antifungals; indicating the need of further research.

1 illus, 94 ref

030332   TEJAN N, DATTA P, GUPTA V (Microbiology Dep, SGPGIMS, Lucknow - 226 014, Email: getnids@gmail.com) : Bacterial diarrhoea: A comprehensive review. Int J Pharm Sci Res 2018, 9(12), 5015-31.

Infective diarrhoea is a common cause of malnutrition in children < 5 years of age. The aetiological agents may be bacteria, viruses or parasites. The bacterial agents cause diarrhoea by either secretion of toxins which act on the small intestine to cause outpouring of fluids into the lumen as seen in Vibrio cholerae, enterotoxigenic Escherichia coli, Clostridium perfringens, Staphylococcus aureus and Bacillus cereus or by damaging mucosa resulting in dysentery like that seen in infection with Shigella, non-typhoidal Salmonellae, Vibrio parahemolyticus, Clostridium difficile and Campylobacter. This article summarizes the common bacterial etiological agents, clinical presentation of illness caused the laboratory diagnosis and antibiotic susceptibility testing of these pathogens.

2 tables, 126 ref