Milk somatic cell DNA isolation and characterization of κ-casein gene in Halari donkey milk.

Halari donkey breed is one of the indigenous breeds of India and its population is rapidly decreasing. The Jenny milk is more similar to human milk, exhibits a wide range of probiotic diversity and hypo-allergenicity, especially among infants suffering from cow and buffalo milk protein allergy. Some studies indicated low levels of κ-casein fraction of casein protein in donkey milk. The k-casein gene was not amplified from the DNA derived from the milk somatic cells of Halari donkeys. The Halari donkey milk has low somatic cells count. We report the first isolation of DNA from milk somatic cells of Halari donkeys with subsequent characterization of k-casein gene. Through our work, we showed that the milk somatic cells can be used as a non-invasive source for DNA isolation towards molecular studies. It also proved the presence of k-casein gene in Halari donkey milk by its amplification from isolated DNA.

Identification and characterization of single nucleotide polymorphisms in DMRT3 gene in Indian horse (Equus caballus) and donkey (Equus asinus) populations.

Equines' ability in racing and riding as well as gaitedness have influenced the human civilization. Aim of this study was to identify and characterize the novel polymorphisms or SNPs in DMRT3 gene in Indian horse and donkey breeds. In this study, the DMRT3 gene was sequenced and characterized in 72 Indian horses' and 33 Indian donkeys' samples. One SNP (A > C) at 878 was found in studied horses while identical SNPs (A > C) at two different nucleotide positions i.e., 878 and 942 in DMRT3 gene (chromosome 23) were observed in studied Indian donkey breeds. Horses and donkeys both have a non-synonymous mutation (A > C) at nucleotide 878 (codon 61) that converts a Stop codon (TAG > TCG) to coding codon Serine, whereas donkeys have a synonymous mutation at nucleotide 942 (codon 82) that converts Serine (TCA > TCC) into Serine. A phylogenetic tree indicated that the DMRT3 gene was equally distributed among the equine breeds. Most of the donkey breeds have been shown high levels of genetic diversity while horse breeds and Halari donkey showed the least genetic diversity. Mutation in DMRT3 has a major impact on gaitedness in horses and is presented at a high frequency in gaited breeds and in horses breed for harness racing.

Molecular regulation of conditioning film formation and quorum quenching in sulfate reducing bacteria.

Sensing surface topography, an upsurge of signaling biomolecules, and upholding cellular homeostasis are the rate-limiting spatio-temporal events in microbial attachment and biofilm formation. Initially, a set of highly specialized proteins, viz. conditioning protein, directs the irreversible attachment of the microbes. Later signaling molecules, viz. autoinducer, take over the cellular communication phenomenon, resulting in a mature microbial biofilm. The mandatory release of conditioning proteins and autoinducers corroborated the existence of two independent mechanisms operating sequentially for biofilm development. However, both these mechanisms are significantly affected by the availability of the cofactor, e.g., Copper (Cu). Generally, the Cu concentration beyond threshold levels is detrimental to the anaerobes except for a few species of sulfate-reducing bacteria (SRB). Remarkably SRB has developed intricate ways to resist and thrive in the presence of Cu by activating numerous genes responsible for modifying the presence of more toxic Cu(I) to Cu(II) within the periplasm, followed by their export through the outer membrane. Therefore, the determinants of Cu toxicity, sequestration, and transportation are reconnoitered for their contribution towards microbial adaptations and biofilm formation. The mechanistic details revealing Cu as a quorum quencher (QQ) are provided in addition to the three pathways involved in the dissolution of cellular communications. This review articulates the Machine Learning based data curing and data processing for designing novel anti-biofilm peptides and for an in-depth understanding of QQ mechanisms. A pioneering data set has been mined and presented on the functional properties of the QQ homolog in Oleidesulfovibrio alaskensis G20 and residues regulating the multicopper oxidase properties in SRB.

Recent Update on Active Biological Molecules in Generating the Anticancerous Therapeutic Potential of Garcinia mangostana.

The Garcinia mangostana Linn (Mangosteen) is also called as "Queen of Fruits" in Malaysia. It is found in the region of Southeast Asia. It is a medicinal plant that has been used to treat cancer in a variety of cell lines. The mangosteen pericarp possesses distinctive biological properties like anticancer or antitumoral and antioxidant. It has a distinct sweet and sour taste, rich in biological compounds like xanthones. It exhibits various properties like apoptotic in tumor cells which leads to the suppression of their growth and results in their various sizes. The primary purpose of this review article is to summarize the valuable results covered by the researchers so far in the Garcinia mangostana extract and its compound like xanthones. Our focus was to explain the role of the phytoconstituent molecules in invading the cancer pathways to combat the expansion of cells. Furthermore, we still feel that there is a scope for more in silico and in vivo studies to understand and identify the specific site of action in tumoral cells and their mechanistic pathways. In conclusion, Garcinia mangostana can act as an anticancer agent by attacking various molecular pathways.

Characterization of Partial Sequence of Myostatin Gene Exon 2 along with SNP detection in Indian Horse Breeds (Equus caballus).

India has well documented horse and pony breeds; however, the population is well diversified in different geographical regions. The Myostatin gene is one of the most profoundly studied genetic components for the detection of SNP's for the performance analysis in horses. In the present study, the MSTN exon 2 partial cds were amplified, sequenced and characterized in about 60 samples of eight different breeds of Indian horses. The results indicated the transition of Thymine to Cytosine (T>C) as single nucleotide polymorphisms in the partial sequence of exon 2 of the MSTN gene at two different codon positions (T12C, T13C) on chromosome 18. The haplotypes and phylogeny of the MSTN gene in the selected horse population were also analyzed. The overall and singleton haplotype are two different entities, indicating the variation among breeds is unique while the gene is equally distributed throughout the population. The phylogeny suggests that all the breeds are somehow equally distributed in their specific geographical tracts. It is the first study of MSTN gene variations in Indian horse breeds, which provides insight into predicting athletic performance as well as phylogeny. This study provides useful genetic information on Indian horses that can be used to model the racing performances of the breeds.

Occupational health hazards and wide spectrum of genetic damage by the organic solvent fumes at the workplace: A critical appraisal.

Long-term exposure to organic solvents is known to affect human health posing serious occupational hazards. Organic solvents are genotoxic, and they can cause genetic changes in the exposed employees' somatic or germ cells. Chemicals such as benzene, toluene, and gasoline induce an excessive amount of genotoxicity results either in genetic polymorphism or culminates in deleterious mutations when concentration crosses the threshold limits. The impact of genotoxicity is directly related to the time of exposure, types, and quantum of solvent. Genotoxicity affects almost all the physiological systems, but the most vulnerable ones are the nervous system, reproductive system, and blood circulatory system. Based on the available literature report, we propose to evaluate the outcomes of such chemicals on the exposed humans at the workplace. Attempts would be made to ascertain if the long-term exposure makes a person resistant to such chemicals. This may seem to be a far-fetched idea but has not been studied. The health prospect of this study is envisaged to complement the already existing data facilitating a deeper understanding of the genotoxicity across the population. This would also demonstrate if it correlates with the demographic profile of the population and contributes to comorbidity and epidemiology.

Cross Talk Between Renal Transporters and Polycystin-1 as a Potential Molecular Target Involved in Autosomal Dominant Polycystic Kidney Disease.

INTRODUCTION: The mutational changes in Polycystin-1(PC-1) encoded by PKD1 gene is the main cause of Autosomal Dominant Polycystic kidney disease (ADPKD). The pathological changes in renal epithelial cells and multiple cyst formation occur due to activation of cascade of signalling pathways and membrane renal transporters (RTs). Our study have focused on the identification, of different RTs, their interactions with Polycystin-1 and other selected target proteins to find out their role in pathogenesis. METHODS: In this study, various RTs protein sequences were identified and retrieved from NCBI's GenBank and UniProt. RTs were categorized according to different nephronal segmenta as per their functional information retrieved from UniProt and Transpoter databases. Further, sequences were subjected for interaction network analysis in String database and Cytoscape 3.7.2. Different interactions including experimentally validated were identified and can be further validated through in vivo methods. RESULTS: The cross talk between different RT, Polycystin-1 and other sequences were analysed. The various pathways of the interaction with PC-1 were categorised. The total number of 119 nodes and 769 edges interactions were generated. The results were visualized and cross verified with other databases in cytoscape. CONCLUSION: The cross signalling of PKD1 with SCNN1A, SCNN1G, SLC12A1, AVPR2 shows their importance in the cyst formation and in pathogenesis of ADPKD.

Polymorphic variation of CYP1A1 and CYP1B1 genes in a Haryana population.

Cytochrome P450 (CYP) 1A1 and CYP1B1 are important phase I xenobiotic metabolizing enzymes involved in the metabolism of numbers of toxins, endogenous hormones, and pharmaceutical drugs. Polymorphisms in these phase I genes can alter enzyme activity and are known to be associated with cancer susceptibility related to environmental toxins and hormone exposure. Their genotypes may also display ethnicity-dependent population frequencies. The present study was aimed to determine the frequencies of commonly known functional polymorphisms of CYP1A1 and CYP1B1 genes in a Haryana state population of North India. The allelic frequency of CYP1A1 polymorphism m1 (MspI) was 29.65% and m2 (Ile(462)Val) was 24.85%. The frequency of CYP1B1 polymorphism m1 (Val(432)Leu) was 45.85% and m2 (Asn(453)Ser) was 16.2%. We observed inter- and intra-ethnic variation in the frequency distribution of these polymorphisms. Analysis of polymorphisms in these genes might help in predicting the risk of cancer. Our results emphasize the need for more such studies in high-risk populations.

CYP1A1 gene polymorphisms: modulator of genetic damage in coal-tar workers.

AIM: It is well known that polycyclic aromatic hydrocarbons (PAHs) such as benzo (a) pyrene have carcinogenic properties and may cause many types of cancers in human populations. Genetic susceptibility might be due to variation in genes encoding for carcinogen metabolizing enzymes, such as cytochrome P-450 (CYP450). Our study aimed to investigate the effect of genetic polymorphisms of CYP1A1 (m1 and m2) on genetic damage in 115 coal-tar workers exposed to PAHs in their work place. METHODS: Genetic polymorphisms of CYP1A1 were determined by the PCR-RFLP method. Comet and buccal micronucleus assays were used to evaluate genetic damage among 115 coal tar workers and 105 control subjects. RESULTS: Both CYP1A1 m1 and CYP1A1 m2 heterozygous and homozygous (wt/mt+mt/mt) variants individually as well as synergistically showed significant association (P<0.05) with genetic damage as measured by tail moment (TM) and buccal micronuclei (BMN) frequencies in control and exposed subjects. CONCLUSION: In our study we found significant association of CYP1A1 m1 and m2 heterozygous (wt/mt) +homozygous (mt/mt) variants with genetic damage suggesting that these polymorphisms may modulate the effects of PAH exposure in occupational settings.

Expression of transcriptional factor genes (Oct-4, Nanog, and Sox-2) and embryonic stem cell-like characters in placental membrane of Buffalo (Bubalus bubalis).

The aim of the study was to assess the expression of transcriptional factor genes and embryonic stem cell-like characters in the placental membrane of buffalo (Bubalus bubalis). Along with the placenta, amniotic fluid, maternal peripheral blood, and umbilical cord blood samples were taken for the future study. The isolation and culture of cells from the placental membrane was followed by the determination of RT-PCR-based markers (Oct-4, Nanog, Sox-2, alkaline phosphatase, stem cell factor, and Nestin) of these cells. Placental membrane cells also positively expressed alkaline phosphatase staining. We isolated adherent cells from trypsin-EDTA-digested placentas and examined these cells for morphology, surface markers, and differentiation potential and found that they expressed several stem cell markers. They also showed neurogenic and adipogenic differentiation potentials under appropriate guided conditions. We suggest that placenta-derived cells have multilineage differentiation potential similar to mesenchymal stem cells in terms of morphology and cell-surface antigen expression. The placenta may prove to be a useful source of mesenchymal stem cells.

Effect of genetic polymorphism of GSTM1 and GSTT1 genotypes on cytogenetic biomarkers among coaltar workers.

Chromosomal aberrations (CAs) in peripheral blood lymphocytes and micronuclei (MN) in exfoliated buccal cells have been used for decades as cytogenetic biomarkers to investigate genotoxicity among occupationally or environmentally exposed population. In our study, we investigated the association of increased cytogenetic damage with genetic polymorphism in glutathione-S transferase genotypes among occupationally exposed 115 coaltar workers and 105 unexposed controls. We found higher mean value of chromosome aberrations (chromatid type-2.01±1.76; chromosomal type-2.22±1.73) and buccal micronuclei (BMN-7.10±1.56) in exposed subjects when compared to referents (chromatid type-0.82±.51; chromosomal type-0.87±.54; BMN-5.09±2.88). We observed that individuals having null genotype of GSTM1 and GSTT1 have significantly higher frequency of CAs and MN. Despite of small sample size, our findings suggest a significant association between polymorphism of glutathione-S transferase genotypes and cytogenetic biomarkers which are considered as early effects of genotoxic carcinogens.

Association of GSTM1 and GSTT1 polymorphisms with DNA damage in coal-tar workers.

DNA damage was evaluated by alkaline comet assay in peripheral blood lymphocytes of 115 coal-tar workers occupationally exposed to polycyclic aromatic hydrocarbons (PAHs) and 105 control subjects. The effect of polymorphisms of glutathione S-transferase (GST) genotypes on the DNA damage was assessed. The mean tail moment (TM) value in the coal-tar workers was significantly higher as compared to the control subjects (12.06 ± 0.55 versus 0.44 ± 0.31; P<0.05). No significant association (P>0.05) between the GSTT1 and GSTM1 genotypes and the TM values was found, however highest mean rank TM value was reported in GSTM1 null and GSTT1 null genotypes in both control and exposed subjects. Our results suggest that there is increased DNA damage in coal-tar workers due to PAHs exposure. Polymorphisms in GSTM1 and GSTT1 genes do not show significant effect (P>0.05) on DNA damage.

Influence of GSTM1 and GSTT1 genotypes and confounding factors on the frequency of sister chromatid exchange and micronucleus among road construction workers.

In the present study, we have investigated the influence of polymorphism of GSTM1 and GSTT1 genes and confounding factors such as age, sex, exposure duration and consumption habits on cytogenetic biomarkers. Frequency of sister chromatid exchanges (SCEs), high frequency cell (HFC) and cytokinesis blocked micronuclei (CBMN) were evaluated in peripheral blood lymphocytes of 115 occupationally exposed road construction workers and 105 unexposed individuals. The distribution of null and positive genotypes of glutathione-S transferase gene was evaluated by multiplex PCR among control and exposed subjects. An increased frequency of CBMN (7.03±2.08); SCE (6.95±1.76) and HFC (6.28±1.69) were found in exposed subjects when compared to referent (CBMN - 3.35±1.10; SCE - 4.13±1.30 and HFC - 3.98±1.56). These results were found statistically significant at p<0.05. When the effect of confounding factors on the frequency of studied biomarkers was evaluated, a strong positive interaction was found. The individuals having GSTM1 and GSTT1 null genotypes had higher frequency of CBMN, SCE and HFC. The association between GSTM1 and GSTT1 genotypes and studied biomarkers was found statistically significant at p<0.05. Our findings suggest that individuals having null type of GST are more susceptible to cytogenetic damage by occupational exposure regardless of confounding factors. There is a significant effect of polymorphism of these genes on cytogenetic biomarkers which are considered as early effects of genotoxic carcinogens.