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Introduction: Male infertility (MI) in about 50% of couples is constituted due to genetic, hormonal, and epigenetic factors. Almost 10% of it is contributed to genetic anomalies. Objectives: This study was undertaken in 787 males with infertility and the impacts of the type of translocations in these cases were studied in relation to MI. Materials and Methods: Five milliliters (ml) of blood were drawn from referred cases suspected with MI and were used for cytogenetic analysis with Giemsa stain following International System for Human Cytogenetic Nomenclature. Results: Of 6142 referral cases, 787 were detected with MI (787/6142; 12.8%). Forty-four translocations (44/787; 5.6%) were detected in 787 infertile men. Other chromosomal abnormalities were 21% (167/787) in them. Among all translocations, an autosome-autosomal translocations were higher in frequency (24/787; 3.1%) than Robertsonian (15/787; 1.9%) and autogonosomal translocations (5/787; 0.6%). Few translocations seemed to be rare as our knowledge is concerned. Maharashtra had high frequency of translocations followed by Delhi union territory (UT) and Gujarat (07 each) and other states. Most of the translocations were related to infertility. Seven males had primary infertility (7/44) and six with secondary type (6/44) in this study. Conclusions: These reciprocal translocations are important in causation of infertility affecting testicular spermatogenesis. Few are rare in our study. Such cases are clinically suggested for counseling before adopting assisted reproductive technologies.

Hydatid disease is caused by the adult and the larval stage of tapeworms belonging to the Echinococcus species. The major sites of its infection are the liver and lungs. Rarely, it affects the pelvic region including the ovary, accounting for 0.2–2.25%. The primary ovarian site is a rare finding. We report here a case series of two cases with hydatid cysts as the left ovarian and tubo-ovarian masses. In case 1, a 30-year-old woman complained about lower abdominal pain with swelling and had a 7-month-old child and amenorrhea. Ultrasonographic report exhibited multiple cysts in the ovary. In case 2, a 23-year-old female was having primary left ovarian mass and clinically reported an ovarian dermoid tumor after ultrasound scan and had abdominal lump with pain. Her vital indices were normal. The biopsy was sent for further investigation.

Background: Y chromosome microdeletions provide a pivotal role in the control of spermatogenesis. These are located on the q arm of it, specifically, the azoospermia factor (AZF) region, hence named as Yq microdeletions. The mutations in this region are related to testicular pathologies such as azoospermia, oligozoospermia, and other semen categories. Objectives: This study was undertaken to review these microdeletions, screening, and their correlation to semen categories in relation to the Indian and Global populations in context to male infertility, future strategies, and its implications. Methodology: These deletions are screened using sequence-tagged sites (STS) of the European Academy of Andrology (EAAs) and Non-EAAs with polymerase chain reaction technology. The data included were Indian and Global studies obtained from research articles, abstracts, and reviews (75) indexed in Google Scholar, PubMed, and Scopus. Results: A number of total 45,562 Y chromosomes were analyzed. In India, this frequency contributes to 8.33% (453/5435) against the average frequency of 6.96% (3170/ 45562) worldwide. Globally, this frequency was higher (9.1%) in the North American continent. Among all types of deletions, AZFc deletions are higher followed by b, a, b+c, and others as well as are related to azoospermia than oligospermia and other semen cases/categories globally including India. In India, the data on partial deletions are scanty. Further, cases with AZFc are better suitable for assisted reproductive technologies after counseling. Conclusion: Evaluation of correct deletion type to a specific testis pathologic phenotype is suggested for sperm retrieval to correct the male partner. This study is hence better suitable for azoospermia with AZFc deletion around the Global and Indian scenario in association with male infertility. Future strategies are thus called for successful treatment of a specific microdeletion clinically in the male.

Background: Y chromosome microdeletions are one of the genetic causes of male infertility. Evaluation of this is important clinically to assist couple with such genetic defects using polymerase chain reaction-based sequence-targeted-site (STS). Objective: The objective of the study was to evaluate Yq microdeletion in suspected infertile male patients referred to our genetic center. Materials and Methods: European Association of Andrology STS markers for azoospermia factor (AZF) region were used to analyze these anomalies. In addition to semen analysis, cytogenetic, and hormonal studies have also been carried out using respective techniques. Results: Three of 41 cases (7.3%) included in our study had Yq microdeletion, of which one case was of azoospermia and two cases of severe oligozoospermia in AZFa and AZFc subregions were detected. Hormonal levels and karyotypes were unchanged. Conclusion: Yq microdeletion in our study exhibited is similar to Maharashtra, but it varies region-wise and ethnicity, including across Indian populations. Such couples require counseling before opting assisted reproductive technologies.

Introduction: Infertility affects about 15% of couples attempting pregnancy and in approximately 50% of these cases, male factors are responsible. Male infertility is clinically characterised by azoospermia and oligozoospermia depending on the amount of loss of genetic material and the size of the affected region on the Y Chromosome Microdeletions (YCM). The majority of genes located in the Y chromosome are involved in male related functions such as spermatogenesis in human, in addition to other endocrine and physiological factors. These microdeletions are located on q arm of Y chromosome, specifically Azoospermia Factor (AZF) region, hence called Yq microdeletions. These deletions are in form of complete/ incomplete, recombination; mutations and Copy Number Variations (CNV) and vary in frequency depending on region, ethnicity, lifestyles and other epigenetic factors. Hence, this study is well reviewed in Indian men with infertility caused by AZF a,b,c and other partial deletions. So, it is important to the one who is affected by these mutations and infertile couples who adopt Assisted Reproductive Technologies (ARTs) after counseling. It is further useful for prediction of testicular sperm retrieval chances. Aim: To review the current status of Yq microdeletion frequency in infertile Indian men with the available data and their correlation with testicular phenotypes as well as other factors. These would also reckon as a supportive to other clinical findings for diagnosis of specific deletion of infertility to adopt ARTs to the infertile couple. Materials and Methods: Various studies including our data were collected to European Molecular Genetics Quality Network (EMQN) as well as analyse these Yq microdeletions screened using specific Sequence Tagged Sites (STS) of available kit like European Academy of Andrology (EAA) and non-EAAs using Polymerase Chain Reaction (PCR) technology. Various researchers from various zones of India contributed to microdeletion screening of Y chromosome using various STS to AZF locus. These data from 30 study groups were compared to geographical areas/zones, Indian populations, environment, selection criteria and other factors in this review. Results: The data on thousands of Y chromosome analysis confirmed that the frequency of microdeletions are affected by sample size, selection criteria of subjects, different geographical regions, ethnicity, Oxidative Stress (OS), Deoxyribonucleic Acid (DNA) fragmentation and food styles in addition to genetic defects. In Indian subcontinent, these deletions contribute to 8.33% from screening of 5435 Y chromosomes (453/5435). Lower percent (5.37%) of Yq microdeletions in Western India than other parts was observed, being highest in South East (20.52%) and North East zones (17.77%) as mentioned in the present study. These variations in Yq microdeletions are attributed to geographic region, foodstyle, other environmental factors and others. AZFc deletions were more prevalent and correlated to azoospermia (referred/ selected, 66%; deleted 61%) from 30/15 citations respectively in present cohort over oligospermic and/or severe oligospermic men followed by b and a sub-regions including b+c, a+b and others in AZF locus. Amongst 30 study groups, 27 exhibited AZFc deletions at higher rate. Conclusion: From these data in India, it was hence noticed that screening of Yq microdeletion is an important criterion and its correlation with spermeograms is very necessary to infer degree of infertility in men. Such cases are strongly suggested to undergo genetic counselling before adoption of ARTS as deletions increase risk of genetic anomalies, low birth weight and congenital malformations in New Births (NB) of Intracytoplasmic Sperm Injection and Testicular Sperm Ejaculates (ICSI/TESE) adopted cases. Thus, Y deletion evaluation reckons the diagnosis of type of male infertility and its prevention in the next generation propagation through ARTs adopting infertile couples after counselling.

Introduction: Cytomegalovirus (CMV) infection in immuno-compromised bone marrow transplant recipients is known. Its infection is about 90% in Indian population, but its symptoms in various age groups in different blood components are scanty and variable. Aim: To evaluate the Whole Blood (WB) and Plasma (PL) samples in population of adult and paediatric leukaemia bone marrow recipients using real-time PCR for CMV infection monitoring. Materials and Methods: In the present observational study, total of 1021 allogeneic bone marrow transplant recipients from our referral centre, including 90 adults and 171 children (paediatric) with follow-up cases were analysed for viral loads in all cases from March 2017 to December 2018. A suitable RT PCR assigned to amplify the conserved region of the UL132 gene to study with respect to two biological components (WL & PL) was investigated in all the patients. Results: The results for CMV infection analysed on both PL and WB showed a good correlation. The kinetics of CMV DNA viral load within the same patient in both WB and PL showed no significant difference. In 2112 follow-up low level of CMV DNA persisted in WB (4.78%), while it was absent in PL. Conclusion: The use of CMV PCR for WB and PL is appropriate to guide implementation and discontinuation of therapy to CMV might lower the risk of CMV disease when given accurate treatment. It might also assist reduce the side effects and duration of treatment. Detection of residual viremia in WB and PL might also offer comparable results in our study cohort.

Background: Hetero atom containing compounds are well studied class of organic compounds exhibits variety of properties and applications. Design and synthesis of new heterocyclic compounds are always of great interest in synthetic and medicinal organic chemistry. Benzothiazole or 2-aminobenzothiazole scaffold based derivatives were reported to display a wide range of biological activities including anticancer, anti-tubercular, antiviral, fungicidal, etc. On the other hand, 1,3,4-oxadiazoles were permit to increase their biological activities due to H-bonding with receptors. These derivatives possess diverse biological activities which include anticancer, antiviral, antifungal, antibacterial and antidepressant etc. Due to interesting biological activity information of about these hetero cyclic moieties, benzothiazole/2-aminobenzothiazole and 1,3,4-oxadiazoles moieties, we chose to design a new series of heterocyclic compounds by mimicking these two types of scaffolds in a single molecule for our study. Methods: The 1,3,4-oxadiazole linked benzothiazole derivatives were synthesized by condensation of, 2-(4-(5-(benzo[d]thiazol-2-yl)-1,3,4-thiadiazol-2-yl)-2,6-dimethoxyphenoxy)acetohydrazide and POCl3 under reflux conditions. All these ten compounds structures were confirmed by spectral data 1H & 13C NMR, Mass, CHN analysis etc. Further, these compounds were evaluated for their anticancer activity against four human cancer cell lines, A549, MCF7, A375 and HT-29 in comparison to CA4 as a reference drug. We also carried out docking studies of these compounds in the Colchicines binding site of Tubulin (PDB_ID: 1SA0) using Glide docking tool indicated that the ligands show good interactions with active site residues. Results: A new series of 1,3,4-oxadiazole fused benzothiazole derivatives were synthesized successfully in totally six steps starting with 4-hydroxy-3,5-dimethoxybenzoyl chloride. All these newly synthesized compounds structures were confirmed by spectral studies and elemental analysis. As we designed for anticancer activity, they were assessed for their anticancer activity against four human cancer cell lines in comparison to a reference drug CA4. As expected, all the ten compounds exhibited anticancer activities against four cancer cell lines with half maximal inhibitory concentration (IC50) values ranging from 0.01 µM to 12.3 µM. The docking studies indicated all the compounds exhibited good binding energies with the receptor. Conclusion: In this study we designed a new series heterocyclic compounds by mimicking two types of scaffolds benzothiazole/2-aminobenzothiazole and 1,3,4-oxadiazoles moieties in a single molecule based on their biological activity in the literature. They were synthesized successfully and molecular structures were confirmed by spectral studies. As expected, all the compounds exhibited anticancer activities against four cancer cell lines. This study can provide a roadmap for design and synthesis of new drug molecules for antitumor and anticancer activity.

Melatonin (MLT) is an extraordinary antioxidant, which plays an important role in reducing reactive oxygen species (ROS) by scavenging them directly or indirectly. Mercury (Hg) is a heavy metal, which induces cytogenetic alterations via various mechanisms, leading to genotoxicity. It induces genotoxicity by enhancing the ROS chiefly. In the present study, the antigenotoxic effect of MLT was evaluated against mercuric chloride (HgCl2). All experiments were conducted in vitro in peripheral blood lymphocytes. Blood cultures were exposed to various concentrations of HgCl2 (2.63, 6.57, and 10.52 microM) for 24 h to study a range of genotoxic parameters. MLT (0.2 mM) supplementation, alone and in combination with the high concentration of Hg, was administered to blood cultures for 24 h. Genotoxic parameters, such as chromosomal aberrations (CAs; structural aberrations (chromatid gaps and breaks, chromosomal gaps and breaks) and numerical aberrations), micronuclei (MNs), and comet assay, were evaluated and analyzed using suitable statistical analysis. Hg treatment revealed a significant increase in CAs, MNs, and comet length. Co-supplementation of MLT along with Hg showed marked protection of these genotoxic end points in treated cultures. In conclusion, our findings suggest that MLT protects against Hg-induced augmentation in genotoxic indices because of its antioxidant property.

Nidhi D Shah,1 Parth S Shah,2 Yash Y Panchal,3 Kalpesh H Katudia,3 Nikunj B Khatri,3 Hari Shankar P Ray,3 Upti R Bhatiya,3 Sandip C Shah,3 Bhavini S Shah,1 Mandava V Rao4 1Department of Pediatrics, Nassau University Medical Center, New York City, NY, USA; 2Department of Medicine, Lahey Hospital and Medical Center, Boston, MA, USA; 3Supratech Micropath Laboratory and Research Institute, Ahmedabad, Gujarat, India; 4School of Sciences, Gujarat University, Ahmedabad, Gujarat, India Background: Germline mutations BRCA1 and BRCA2 contribute almost equally in the causation of breast cancer (BC). The type of mutations in the Indian population that cause this condition is largely unknown.Purpose: In this cohort, 79 randomized BC patients were screened for various types of BRCA1 and BRCA2 mutations including frameshift, nonsense, missense, in-frame and splice site types. Materials and methods: The purified extracted DNA of each referral patient was subjected to Sanger gene sequencing using Codon Code Analyzer and Mutation Surveyor and next-generation sequencing (NGS) methods with Ion torrent software, after appropriate care. Results: The data revealed that 35 cases were positive for BRCA1 or BRCA2 (35/79: 44.3%). BRCA2 mutations were higher (52.4%) than BRCA1 mutations (47.6%). Five novel mutations detected in this study were p.pro163 frameshift, p.asn997 frameshift, p.ser148 frameshift and two splice site single-nucleotide polymorphisms (SNPs). Additionally, four nonsense and one in-frame deletion were identified, which all seemed to be pathogenic. Polymorphic SNPs contributed the highest percentage of mutations (72/82: 87.8%) and contributed to pathogenic, likely pathogenic, likely benign, benign and variant of unknown significance (VUS). Young age groups (20–60 years) had a high frequency of germline mutations (62/82;75.6%) in the Indian population. Conclusion: This study suggested that polymorphic SNPs contributed a high percentage of mutations along with five novel types. Younger age groups are prone to having BC with a higher mutational rate. Furthermore, the SNPs detected in exons 10, 11 and 16 of BRCA1 and BRCA2 were higher than those in other exons 2, 3 and 9 polymorphic sites in two germline genes. These may be contributory for BC although missense types are known to be susceptible for cancer depending on the type of amino acid replaced in the protein and associated with pathologic events. Accordingly, appropriate counseling and treatment may be suggested. Keywords: breast cancer, BRCA1 and BRCA2 gene mutations, polymorphic SNPs, novel types, age, NGS, Sanger gene sequencer&nbsp

Several studies documented the ameliorative effects of curcumin which plays a pivotal role in radical scavenging activities. It also participates in various cellular pathways and interacts with multiple targets. In the present study, we investigated the ameliorative effect of curcumin upon chromosomal genotoxicity induced by cyclosporine, an immunosuppressant, using in vitro approaches. A plausible mechanism of how curcumin mitigates the genotoxic implications of cyclosporine was ascertained using in silico tools. We observed that the curcumin reduces the genotoxic consequences made by cyclosporine upon cell cycle checkpoints and associated chromosomal/DNA manifestations. In addition, we presented the mechanistic details of curcumin interaction with various biomacromolecule types using docking experiments which showed that the possible radical scavenging activities can only be emerged by inducing the expression of antioxidant enzymes, supported by available experimental evidences. We anticipate that the induction of antioxidant enzymes by curcumin would activate Nrf2-Keap1 pathway as the plausible mechanism to exert anti-inflammatory response as demonstrated in renal epithelial cells.

The dengue fever is a debilitating arthropod-borne disease caused by one of the four closely related dengue viruses. The symptoms appear 3 – 14 days after mosquito bite and range from mild fever to...

In the present study, the interaction of the calix[4]resorcinarene polyhydrazide (CPH) capped nanoparticles (CPH-AuNps) and (CPH-AgNps) with different amino acids like arginine, aspartic acid, glutamic acid, glycine, leucine, methionine, histidine, tryptophan and cysteine has been studied by using UV–vis and fluorescence spectroscopy. CPH-AuNps and CPH-AgNps were found to be the selective and sensitive fluorescent probe for l -DOPA and histidine, respectively. The minimum detectable limit for both l -DOPA and histidine was found at 10 nM by a facile way of fluorescence quenching i.e. by a turn off mechanism. CPH-AuNps and CPH-AgNps have also been studied for their antioxidant activity and interaction with DNA by UV–vis spectrophotometry. Furthermore, electrophoresis technique confirmed the interaction of ethidium bromide (EtBr) intercalated CT-DNA and S-DNA with CPH-AuNps and CPH-AgNps, respectively. The simplicity, sensitivity and specificity for the detection of amino acids by the CPH-AuNps and CPH-AgNps, indicates that the synthesized nanoparticles can be used as nano biosensor, with a potential prospect in the biomedical analysis.

Parth S Shah,1 Nidhi D Shah,2 Hari Shankar P Ray,3 Nikunj B Khatri,3 Ketan K Vaghasia,3 Rutvik J Raval,4 Sandip C Shah,3 Mandava V Rao5 1Department of Medicine, Lahey Hospital and Medical Center, Boston, MA, 2Department of Pediatrics, Nassau University Medical Centre, New York, NY, USA; 3Supratech Micropath Laboratory and Research Institute, 4Department of Zoology, 5School of Sciences, Gujarat University, Ahmedabad, Gujarat, India Background: β-Thalassemia is the most prevalent genetic disorder in India. Its traits and coinheritance vary from mild to severe conditions, resulting in thalassemia minor, intermediate, and major, depending upon many factors.Purpose: The objective of this study was to identify the incidence of β-thalassemia traits, their coinheritance, and mutations, as well as to support the patients already diagnosed with β-thalassemia in East-Western Indian population for better management.Patients and methods: Seventy-five referral cases for β-thalassemia were analyzed for various β-thalassemia traits, heterozygosity, and homozygosity conditions. Blood phenotypic parameters using cell counter and capillary electrophoresis were investigated. Analyses of eight common mutations of thalassemia in India were carried out using polymerase chain reaction-amplification refractory mutation system, end point polymerase chain reaction, and DNA sequencing methods.Results: Of these (75) referral cases from East-Western Indian region, 68 were positive for β-thalassemia (90.67%). The majority of case types were of β-thalassemia minor (49, 65.33%), followed by HbE traits (6, 8.0%) and β-thalassemia major, including heterozygous and homozygous (5, 6.66%; 4, 5.33%) types and then HbE homozygous (2, 2.66%), as well as one each of the HbE/β-thalassemia and HbD/β-thalassemia (1, 1.34%) combination. Mutation analysis also revealed that the highest frequency of mutation was c.92+5G>C (41, 60.29%) followed by deletion 619bp (9, 13.23%) and c.79G>A (8, 11.76%) in our study group. Five cases (nos. 24, 27, 33, 58, and 71) exhibited coinheritance between β0/β+ (2), β0/β D (1), and c.124_127delTTCT/β+ or β0 (2) affecting the Rajasthani and Gujarati populations in our study of the Western region of India.Conclusion: We strongly recommend these Western populations for genetic screening before adopting reproductive technologies and interracial marital relations. Keywords: β-thalassemia traits, coinheritance, hematogram, capillary electrophoresis, PCR-ARMS, DNA sequencing, mutation analysis, East-Western India

A report in a Gujarati family in Western India consisting of rare co-inheritance of β-thalassemia (β0/β+) in a proband, son was identified. The trio samples, parents and son, of extracted D...

Background and Objectives: Aneuploidy is one of the major concerns to cause genetic anomalies. This condition is mostly related to addition and/or deletion with respect to set(s) of chromosomes. Here, we report an analysis of 5740 referral cases during three consecutive years (2015 – 2018) from our Diagnostic Research Center, Ahmedabad for aneuploidy pattern. Methodologies: The patients were asked to fill the necessary forms and their blood (5ml) was drawn for chromosomal studies using the Karyotyping following International System for Human Cytogenetic Nomenclature (ISCN) manual. Results: The data revealed the numerical aberrations for only aneuploidy detected was (3.7%; 211/5740). In this report, constitutional (c) autosomal aneuploidy was 75% (158/211). The total mosaic cases were nine (9/211) comprising constitutive (2) and acquired (7) aneuploidy cases. In autosomal aneuploidy, cT21 was higher (96%; 152/158) than others (4%; 6/158) comparatively. Among cT21 (152), males (76%; 115/152) were more affected than females (24%; 37/152). These statistical data also revealed that acquired chromosomal aneuploidy (leukemia) possessed (25%; 53/211); with more mosaic cases (7/211). Conclusion: Couples with such conditions are eligible for genetic tests and counseling as well as new strategies are urgently to be undertaken by governmental organizations (GOs) and non-governmental organizations (NGOs) for affected families with better personalized and informed decision making. The significance of these data is thus discussed in relation to genetic disorders caused by constitutional and acquired aneuploidy of leukemic blood in this report.

A kidney transplant recipient patient was infected by Streptococcal bacterial infection with renal failure. Immnunofloroscent study showed IgA was dominant, but recovered with proper diagnosis and treatment to the 49 year old female in our report from India.

Background and Objectives: Human cell nucleus has, the genome consisting of euchromatin and heterochromatin. The euchromatin has gene-rich and actively functional. The heterochromatin has two components namely constitutive and facultative, where the former is highly polymorphic. It is related to numerous diseases like cancer and infertility which is now well known, though it was earlier thought to be inactive; hence the implication of these polymorphic variants of chromosomes is reviewed with respect to acrocentric and non- acrocentric types. Methodologies: The polymorphic variants can be detected by C, G, Q and R banding techniques. We usually follow G band preparation of karyotypes following World Health Organisation (WHO) manuals and their role in cancer and reproduction is reviewed. Review and Conclusion: It is emphasized that most of the p and q arms of 1, 9, 16, D and G groups and X, Y chromosomes exhibited polymorphism which are related to cancerous and infertile conditions in both sexes. Data on few non-acrocentric chromosomes like 2, 4, 8, 10, 12, 18, 19 and 20 are not available. Our review however indicated that the evaluation of specific heteromorphic variants needs to be detected using specific probes for confirmation of anomaly to assist affected cases, though earlier data indicated ambiguous information with few cases analyzed regarding assisted reproductive technologies and malignancy condition. This appraisal thus would play a key role in human chromosomal heteromorphic abnormalities and recommend genetic tests and counseling ultimately made available to the affected cases.

Introduction: Among all chromosomes (46) in the human genome, particular significance has been given to chromosomes 13, 18, 21, X and Y. This is primarily because of aneuploidy in these chromosomes that result in viable pregnancies with congenital defects. As a result, standardised methods like Rapid Aneuploidy Test (RAT) for detection is the need of the hour in addition to Non-Invasive Prenatal Testing (NIPT) and Chromosomal Microarray (CMA). Aim: To compare and analyse the diagnostic utility of Fluorescent In-Situ Hybridization (FISH) and Quantitative fluorescent Polymerase Chain Reaction (QF-PCR) in aneuploidy of detection. Materials and Methods: In the present observational study, 120 pregnant women suspected of having fetal aneuploidies were subjected to amniocentesis and Chorionic Villus Samplling (CVS). Following DNA extraction, FISH and QF-PCR were carried out using pre-designed chromosomal markers and specific FISH probes for trisomy of 13, 18 and 21. Results: Of 120, 5 prenatal samples showed Trisomy (T) 13, 18 and 21 chromosomes, amounting to a frequency of 4.2% (5/120). These results were concordant by both tests i.e FISH and QFPCR trisomy 18 and 21 detected. Four amniotic fluid samples, two each respectively (4/108; 3.7%), and one Chorionic Villus Sampling (CVS) (1/12; 8.3%) were tested positive for trisomy of chromosome 13. Conclusion: From the present study, it can be concluded that QF-PCR is a better technique for detection of aneuploidies. However, both these techniques, together called RAT of Invasive Prenatal Screening (IPS) should be performed for errorless results before termination of pregnancy (TOP).

Present study represents a novel approach of PCR ready-human DNA extraction method from skeletal remains using copper nanoparticles (CuNPs) for personnel identification. To achieve rapid, cost effective, sensitive and non-hazardous method for DNA extraction we utilized CuNPs synthesized using microwave. The applicability of this approach was first tested in blood samples and afterwards, this system was extended to skeletal remains' samples also. This method yields good quality DNA that are ready for PCR reactions from small quantities of blood and skeletal remains. Consequently, even small quantities of nanoparticles could be potentially utilized for a highly efficient isolation of DNA from skeletal remains as well as from ancient archaeological samples. The present method has the advantages that it is quick with high yield, inexpensive, robust, environment friendly and does not require use of hazardous organic solvents.

A recurring problem in forensic medicine is the need to fix the time of death within the limits of probability. Errors in overestimating and underestimating the post-mortem interval based on body cooling is common. Post-mortem biochemistry has become a potent additional procedure in forensic death investigation. However, in our experiment, to determine the post-mortem interval we investigated the ATPase activity. The investigation was performed on the liver tissue of 25 corpses (21 men and four women) of age ranging between 40–60 years. The study of ATPase enzymatic activity shows that there is a considerable drop in the levels of ATPase activity with increasing post-mortem interval up to 24 h and this is one of the most reliable methods for estimating time of death.

We performed transcriptome sequencing of canine retinal tissue by 454 GS-FLX and Ion Torrent PGM platforms. RNA-Seq analysis by CLC Genomics Workbench mapped expression of 10,360 genes. Gene ontology analysis of retinal transcriptome revealed abundance of transcripts known to be involved in vision associated processes. The de novo assembly of the sequences using CAP3 generated 29,683 contigs with mean length of 560.9 and N50 of 619 bases. Further analysis of contigs predicted 3,827 full-length cDNAs and 29,481 (99%) open reading frames (ORFs). In addition, 3,782 contigs were assigned to 316 KEGG pathways which included melanogenesis, phototransduction, and retinol metabolism with 33, 15, and 11 contigs, respectively. Among the identified microsatellites, dinucleotide repeats were 68.84%, followed by trinucleotides, tetranucleotides, pentanucleotides, and hexanucleotides in proportions of 25.76, 9.40, 2.52, and 0.96%, respectively. This study will serve as a valuable resource for understanding the biology and function of canine retina.

Progressive retinal atrophy (PRA) is one of the major causes of retinal photoreceptor cell degeneration in canines. The inheritance pattern of PRA is autosomal recessive and genetically heterogeneous. Here, using targeted sequencing technology, we have performed exome sequencing of 10 PRA-affected (Spitz=7, Cocker Spaniel=1, Lhasa Aphso=1 and Spitz-Labrador cross breed=1) and 6 normal (Spitz=5, Cocker Spaniel=1) dogs. The high-throughput sequencing using 454-Roche Titanium sequencer generated about 2.16 Giga bases of raw data. Initially, we have successfully identified 25,619 single nucleotide polymorphisms (SNPs) that passed the stringent SNP calling parameters. Further, we performed association study on the cohort, and the highly significant (0.001) associations were short-listed and investigated in-depth. Out of the 171 significant SNPs, 113 were previously unreported. Interestingly, six among them were non-synonymous coding (NSC) SNPs, which includes CPPED1 A>G (p.M307V), PITRM1 T>G (p.S715A), APP G>A (p.T266M), RNF213 A>G (p.V1482A), C>A (p.V1456L), and SLC46A3 G>A (p.R168Q). On the other hand, 35 out of 113 unreported SNPs were falling in regulatory regions such as 3'-UTR, 5'-UTR, etc. In-depth bioinformatics analysis revealed that majority of NSC SNPs have damaging effect and alter protein stability. This study highlighted the genetic markers associated with PRA, which will help to develop genetic assay-based screening in effective breeding.

The mutations in Haemoglobin Beta (HBB) gene, bring about less or no production of Hb β-chain synthesis in affected cases, leading from minor to major types depending on haematological indices. In compound heterozygotic conditions, two traits are involved, in which one parent has HbE trait and the other has β-thalassaemia carrier (trait). Here, we report a family of Rajasthan, West India which had a proband (son) having HbE/ β-thalassaemia a co-inherited compound heterozygosity as revealed by DNA sequencing. It also contained upper levels of HbE with altered Hb and red cell indices showing asymptomatic to symptomatic state requiring blood transfusion periodically. The parents and Chorionic Villus Sampling (CVS) were HbE and β-thalassaemia traits only. Such case is rare in Western India and we recommend this family for genetic counseling and genetic testing before they want reproductive choices in future for better management in a society.

Lead (Pb) which plays a significant role in modern industry is related to a broad range of physiological, biochemical, behavioural and genetical dysfunctions. Its exposure leads to an increased frequency of genetic aberrations in humans. Hence, this study was designed to assess the genotoxic effect of lead acetate at three dosage levels (10, 25 and 50 µg/mL) by employing: the Cytokinesis Block Micronucleus (CBMN) assay and the Comet assay in Peripheral Blood Lymphocyte Cultures. The results of this study revealed an increased level of DNA damage among treated groups. A significant increase in the tail length of comets and other indices was observed at 25 and 50 µg/mL concentrations comparatively. Thus, lead acetate induced single-strand breaks (SSB) and double strand breaks (DSB) in DNA, alkali-labile sites (ALS), oxidative DNA damage as well as DNA-DNA/DNA-protein/DNA-metal cross linking as evidenced by the Comet assay. The chromosome breakage, DNA misrepair, chromosome loss and telomere end fusion were determined by the Micronucleus assay. Micronucleus frequency in treated lymphocytes was significantly higher as compared to controls. Nucleoplasmic bridges increased significantly and Nuclear buds increased at higher two doses only in exposed cultures. Thus, these assays are better indices for lead induced genotoxicity and metal-nucleus interactions.

Leiomyosarcoma (LMS) of the colo-rectum is a rare malignant entity with unfavourable prognosis, and responds poorly to conventional chemo-radiation therapy. Only a few cases of the LMS of colorectum have so far been reported earlier. Here we report two such cases of primary leiomyosarcoma of sigmoid colon and rectum. Due to histological similarity with gastrointestinal stromal tumour (GIST), the LMS cases were misdiagnosed as GIST. This is essentially a pathological diagnosis having a similar histomorphological appearance that of GIST, but a different Immunohistochemistry profile, in contrast to GIST. Immunohistocytochemically LMS are negative for CD34, CD117 and DOG1 and positive for desmin as detected in this report. GIST is curable with surgery followed by imatinib (in intermediate /high-risk patients) while LMS behaves aggressively requiring radical excision of the tumour followed by intensive cytotoxic chemotherapy regimens. Thus it is important to diagnose LMS definitively by immunohistochemical profiling as the prognosis and treatment of these tumours vary from others as mentioned.

A number of 2H-1,3-benzoxazin-4(3H)-one derivatives containing indole or benzofuran moieties were synthesized by using Pd/C-Cu mediated coupling-cyclization strategy as a key step. The o-iodoanilides or o-iodophenol were coupled with 3-{2-(prop-2-ynyloxy)ethyl}-2H-benzo[e][1,3]oxazin-4(3H)-one using 10%Pd/C-CuI-PPh3 as a catalyst system and Et3N as a base to give the target compounds. All the synthesized compounds were tested for their PDE4B inhibitory potential in vitro using a cell based cAMP reporter assay. Some of them showed fold increase of the cAMP level when tested at 30 μM. A representative compound showed encouraging PDE4B inhibitory properties that were supported by its docking results.

This work evaluates cationic intercalation of Capecitabine (Xeloda), an antineoplastic agent in interlayer gallery of Na + -MMT, which was further compounded with Poly ( L -Lactide) to form microcomposite spheres, for oral chemotherapy of breast cancer. The XRD, TGA, SEM, TEM and FT-IR analysis indicated the CAP intercalated in interlayer of MMT and stabilized in the longitudinal monolayer mode by electrostatic interface. No significant alteration in fundamental active properties of CAP was found in the MMT efficacy and data from in vitro study showed controlled drug release pattern >72 h. The efficacy of CAP-MMT hybrid/MPs was also confirmed on Human neuroblastoma cell line (IMR32) by in vitro cell culture experiments evaluating cell cytotoxic and oxidative stress marker indices which showed that intercalated drug and MPs, were as effective as pristine drug. In vivo pharmacokinetics of CAP-MMT hybrids in rats was examined and plasma CAP levels were evaluated. Our findings suggest that CAP-MMT hybrids and MPs can be of considerable value in chemotherapy of malignant neoplastic disease with reduced side effects and clearly demonstrated that MMT not only plays a role as a delivery matrix for drug but also facilitates significant increase in the delivery proficiency.

Lead, a heavy metal and multifaceted toxicant, is well studied for its distribution and toxicity in ecosystem, yet there is no consensus on its amelioration by any synthetic or phytochemical compounds. Curcumin, a known antioxidant and dietary element, is a well-known herb, for its therapeutic uses and having a wide spectrum of its beneficial properties against several adverse effects. Hence, the current study was taken into consideration to evaluate the ameliorative effects of curcumin (3.87 μM, i.e. 1.43 μg/ml) against lead acetate (doses: 10-6 M, i.e. 0.379 μg/ml and 10-4 M, i.e. 37.9 μg/ml, durations: 24 h and 69 h) induced genotoxicity and oxidative stress in human peripheral blood lymphocyte cultures (PBLC). On one hand, antigenotoxic and antioxidative potentials of curcumin against lead were simultaneously evaluated by the array of genotoxicity and oxidative stress indices. The result postulated that lead acetate showed dose- and duration-dependent increase in both genotoxicity and oxidative stress whereas curcumin, when added along with lead acetate, showed the significant amelioration in all genotoxic and oxidative stress-related indices. The study indicated that, due to alteration in antioxidant defense system, there is an adverse genotoxic effect of lead. On the other hand, curcumin, a potent antidote, can protect chromatin material against lead -mediated genotoxicity by balancing the activity of antioxidant defense system.

Lead (Pb) is present in the natural and occupational environment and it is one of the most clinically important heavy metals, because it induces a broad range of physiological, biochemical and genetical dys- functions. Its exposure leads to an increased frequency of genetic aberrations in human beings. Hence, this investigation was conducted to assess the genotoxic effect of lead at three dosage levels. In the present study, the DNA damage was detected by adopting the alkaline single cell gel electrophoresis assay (Comet Assay) in peripheral blood lymphocyte cultures. The result of this study revealed an increased level of DNA damage among treated groups. A significant increase in the tail length of comets and other indices was observed at 25 µg/ml and 50 µg/ml concentrations comparatively. Thus lead acetate induced single- strand breaks (SSB) and double strand breaks (DSB) in DNA, alkali-labile sites (ALS), oxidative DNA damage as well as DNA-DNA / DNA-protein / DNA-metal cross linking as evidenced by the comet assay.

Free radicals are common outcome of normal aerobic cellular metabolism. In-built antioxidant system of body plays its decisive role in prevention of any loss due to free radicals. However, imbalanced defense mechanism of antioxidants and overproduction or incorporation of free radicals from environment to living systems leads to serious damage. It also attacks nervous system resulting in neural-degeneration. In order to evaluate the neurotoxic effect on the brain parts of mercury in our study, oxidative stress indices of enzymatic and non enzymatic components were measured in rats intoxicated with mercury (2 mg and 4 mg/kg body weight) for 60 days to adult rats. Along with gravimetry, tissue burden was also recorded. Alterations in these indices were further supported by ultrastructural studies carried out in the brain as indicated by myelin disintegration, cell organelle alterations and neuronal loss by mercury poisoning. Treatment with the antioxidant melatonin (N-acetyl 5-methoxy tryptamine, 5 mg/kg) prevented mercury exerted toxicity due to its antioxidant property. The pathological changes were also ameliorated in the brain region comparatively to support biochemical profile of brain. Thus, melatonin produced neuroprotection against mercury poisoning in rats.

The present study was aimed to evaluate curcumin as a potential natural antioxidant to mitigate the genotoxic effects of arsenic (As) and fluoride (F) in human peripheral blood lymphocytes. The study was divided into nine groups consisting of negative control, positive control treated with ethyl methane sulphonate (EMS; 1.93 mM) and curcumin control with only curcumin (1.7 microM) in blood culture. As (1.4 microM) and F (34 microM) were added alone as well as in combination, to the cultures, with and without curcumin. Cultures were analysed for chromosomal aberrations (both structural and numerical) and primary DNA damage via comet assay as the genotoxic parameters after an exposure duration of 24h. Results revealed that curcumin efficiently ameliorates the toxic effect of As and F by reducing the frequency of structural aberrations (60%), hypoploidy (50%) and primary DNA damage. In conclusion, curcumin mitigates the genotoxic effects of the two well known water contaminants (As and F) effectively and efficiently at the given concentration in vitro.

The effect of melatonin on the neurotoxicity induced by mercuric chloride was studied. Adult rats were fed orally with two different doses of mercuric chloride (2 mg; 4 mg/kg body weight) to evaluate brain toxicity with respect to cerebral hemisphere, cerebellum, and medulla oblongata regions for 60 days with or without supplementation with melatonin (5 mg/kg body weight) intraperitoneally. The results suggest that the graded doses of mercury elicit the depletion of enzymatic activities, such as adenosine triphosphatase, succinate dehydrogenase, phosphorylase, alkaline phosphatase, acid phosphatase, altered glycogen, total protein, and lipid peroxidation levels in the cerebral hemisphere, cerebellum, and medulla oblongata of the brain, thereby affecting their respective functions. Blood glucose and mercury levels increased, followed by a reduction in body and organ weights. All these effects seemed to be severe in the cerebral hemisphere of the brain. Further affected indices were, to some extent, maintained in the brain of animals cotreated with melatonin, showing its protective role against mercury-exerted neurotoxicity.

Present study investigated the protective role of melatonin (MLT, 5 mg/kg body wt., ip) against the long term effects of mercuric chloride (MC; 2 and 4 mg/kg body wt., po) in the thyroid gland of the rats through certain antioxidative indices like superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione (GSH), catalase (CAT) and lipid peroxidation (LPO), other biochemical parameters such as succinate dehydrogenase (SDH), adenosine triphosphatase (ATPase), acid phosphatase (ACPase) and alkaline phosphatase (ALPase) were also measured. Antioxidative enzymes and other parameters showed a significant reduction while LPO and mercury levels increased significantly in a dose dependent manner in MC treated animals as compared to control groups. Co-treatment with MLT revealed no significant effect on antioxidative and metabolic indices in the thyroid gland of rats. The results of present study thus strongly suggest that mercury affected antioxidant defense system and other metabolic enzymes of thyroid. Co-administration of melatonin exerted a protective effect against mercury induced endocrine toxicity.

A novel water-soluble [60] fullerene-substituted lysine derivative 3 has been synthesized and characterized by elemental analysis, 1H NMR, 13C NMR and FAB-MS. The synthetic procedure involved condensation of Boc-protected lysine with terephthaldehyde followed by 1,3-dipolar cycloaddition reaction with C60 in the presence of sarcosine and finally deprotection of the amino group using trifluoroacetic acid. The synthesized compound 3 exhibited high DNA cleavage efficiency upon visible light irradiation in the presence of NADH.

Ground water contamination of arsenic in drinking water is a burning problem worldwide; especially in West Bengal (India) and Bangladesh. The main endeavor in this study was to assess the role of vitamin A (2.72 μM/7 ml culture), a naturally occurring antioxidant upon arsenic-induced genotoxicity; with respect to chromosomal aberrations (structural and numerical) and micronuclei. Whole blood cultures set for 72 h were exposed to test chemicals for a period of 24 h ahead of harvesting. Arsenic concentrations tested in the present study are 0.36, 0.72 and 1.4 μM/7 ml culture. Mitomycin C (MMC), the direct acting mutagen was used as positive control. Our data indicates that at concentrations tested, arsenic-induced increase in the mean frequencies of these genotoxic indices were diminished by vitamin A, indicating its protective role towards cells from arsenic exerted injury.

This study was undertaken to determine whether vitamin A (vit. A), which was shown to possess antioxidant properties when co-administered with arsenic trioxide (As2 O3) might prevent As-induced detrimental effects. Two doses of As (0.5 or 1 mg kg−1 body weight), alone and in combination with vit. A as retinyl palmitate (0.2 mg kg−1 body weight), were used in this study. Parameters assessed included acetyl cholinesterase (AChE) activity and levels of total proteins, cholesterol and total lipids in cerebral hemisphere (CH) and cerebellum (C) regions of the brain as well as the serum of mice. Significant decreases in AChE activity and levels of proteins together with other parameters were noted in the brain and blood. These effects were significantly mitigated by co-treatment with vit. A. These results emphasize that concurrent supplementation of vit. A significantly protects against As-induced toxicity.