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From patients with a poor prognosis of pregnancy, 1620 embryos generated in vitro and chromosomal analysis was performed on these embryos. The result was yielded in 1596 embryos, out of them 536(34%) were euploid and 1060(66%) carried chromosomal abnormalities. In addition, 92% of embryos with multinucleated cells were diagnosed mosaics whereas the 86% of chromosomal abnormalities were associated to the presence of cytoplasmic concentration. For the derivation of the normal embryonic stem cell (ESC)lines and developmental modelling aneuploid embryos have been used. Genetic diagnosis at the cleavage or blastocyst stage could be partly abnormal because during the preimplantation diploid- aneuploid mosaic embryos was most frequently observed. From a single cell of a particular embryo the chromosomal status of that embryo can be determined, thus the prevalence of mosaicism. Detection of aneuploidy in single cells have been developed recently. After conducting research methods, it was confirmed that aneuploidy is a common feature of human oocytes and preimplantation embryos. The detection of segmental aneuploidy is currently considered problematic for embryo diagnosis and patient counselling, so the data are of great relevance for preimplantation genetic testing. The first major milestone in early mammalian embryogenesis was the formation of a totipotence blastocyst which is capable of implantation. The whole chromosomal abnormalities, or aneuploidy, determines whether the human embryos will arrest or reach the blastocyst stage. Certain embryos can still form blastocyst depending on the type of chromosomal abnormalities and that can be morphologically indistinguishable from chromosomally normal embryos.

The aim of this study was to check effectiveness of drug-eluting stents for prevention of restenosis resulting from neointimal hyperplasia. Twenty-one sirolimus-eluting stents (SESs) and bare metal stents (BMSs; control) were compared by implantation into left and right balloon-injured external iliac arteries of New Zealand White rabbits, respectively. Under general anesthesia and angiography, using 3-mm coronary balloon catheters, stents were successfully deployed. Anticoagulant therapy was not given to any animal. Rabbits were allotted into three groups at 7, 14, and 21 days (7 rabbits in each group). Each stented vessel of sacrificed rabbits was subjected to a resin-embedding technique and methyl methacrylate sections were obtained using a tungsten carbide knife for staining. In histological examination, SESs showed marked reductions in almost all parameters. Morphometric results showed that arteries with SESs had a larger luminal area (P < 0.0001), lower intimal (P < 0.001) and medial (P < 0.001) thickness, and lower intimal index (P < 0.003) as opposed to arteries stented with BMSs. Luminal narrowing in SESs was 0% and reendothelialization was similar in both stented groups (90.48%). SESs showed marked reduction in nonocclusive peristrut fibrin deposition versus BMSs. Thus, the SES was more effective for prevention of restenosis than the BMS in rabbit.

Background: Mesial temporal lobe epilepsy (MTLE) is the most common medically refractory epilepsy syndrome in adults, and hippocampal sclerosis (HS) is the most frequently encountered lesion in patients with MTLE. Premature accumulation of corpora amylacea (CoA), which plays an important role in the sequestration of toxic cellular metabolites, is found in the hippocampus of 50-60% of the patients who undergo surgery for medically refractory MTLE-HS. However, the etiopathogenesis and clinical importance of this phenomenon are still uncertain. The ABCB1 gene product P-glycoprotein (P-gp) plays a prominent role as an antiapoptotic factor in addition to its efflux transporter function. ABCB1 polymorphism has been found to be associated with downregulation of P-gp expression. We hypothesized that a similar polymorphism will be found in patients with CoA deposition, as the polymorphism predisposes the hippocampal neuronal and glial cells to seizure-induced excitotoxic damage and CoA formation ensues as a buffer response. Materials and Methods: We compared five single nucleotide polymorphisms in the ABCB1 gene Ex06+139C/T (rs1202168), Ex 12 C1236T (rs1128503), Ex 17-76T/A (rs1922242), Ex 21 G2677T/A (rs2032582), Ex26 C3435T (rs1045642) among 46 MTLE-HS patients of south Indian ancestry with and without CoA accumulation. Results: We found that subjects carrying the Ex-76T/A polymorphism (TA genotype) had a five-times higher risk of developing CoA accumulation than subjects without this genotype (Odds ratio 5.0, 95% confidence intervals 1.34-18.55; P = 0.016). Conclusion: We speculate that rs1922242 polymorphism results in the downregulation of P-gp function, which predisposes the hippocampal cells to seizure-induced apoptosis, and CoA gets accumulated as a buffer response.

Gangliosides isolated from biological sources are usually detected as sodium/potassium adduct ions and these ions are easily fragmented by dissociation of labile glycosidic bonds in the positive ion mode Mass Spectrometry (MS). A large number of conditions for fragment suppression of these acidic glycolipids by using non-acidic matrices or changing metal additions have been demonstrated. Compared to sodium/potassium adduct ions, the cesium adduct ions suppress the fragmentation of these acidic glycolipids. On the contrary, lithium adduct ions induce the fragmentation, but generate more informative fragment ions of glycolipids than other alkali metal adduct ions in post-source decay, MS/MS as well as MS spectra. To suppress the fragmentation of labile glycosidic bond and generate more informative fragmentation, we have examined and established the optimal condition for the detection of [M+Li]+ of gangliosides (GM3 and GM2) using the matrix-assisted laser desorption/ionization time-of-flight MS by adjusting matrix and alkali metal salt combinations and concentrations.

The transmembrane P-glycoprotein that functions as a drug-efflux transporter coded by ATP-binding cassette, subfamily B, member 1/Multidrug Resistance 1 (ABCB1/MDR1) gene is considered relevant to drug absorption and elimination, with access to the central nervous system. Effects of three ABCB1 single nucleotide polymorphisms (SNPs) in genotypic and haplotypic combination have been evaluated in a south Indian population for risk of pediatric medically refractory epilepsy. The study included age and sex matched medically refractory (N=113) cases and drug responsive epilepsy patients (N=129) as controls, belonging to the same ethnic population recruited from a tertiary referral centre, of Karnataka, Southern India. The genotype frequencies of SNPs c.1236C>T, c.2677G>T/A, and c.3435C>T were determined from genomic DNA of the cases and controls by PCR- RFLP and confirmatory DNA sequencing. 256 normal population samples of the same ethnicity were genotyped for the three loci to check for population stratification. Results indicate that there was no statistically significant difference between allele and genotype frequencies of refractory and drug responsive epilepsy patients. The predicted haplotype frequencies of the three polymorphisms did not show significant difference between cases and controls. The results confirm earlier observations on absence of association of ABCB1 polymorphisms with medically refractory epilepsy.

Developing countries are facing severe bottlenecks in the technological advancement in biotechnology, due to restrictions imposed by patent protected products and protocols. This calls for designing of simple and cost-effective alternatives for the indispensable products like DNA molecular weight markers. We demonstrate a novel, rapid and cost-effective method of making in-house 100bp ladder for routine use. In our method we use a single forward primer and five reverse primers designed on the backbone sequence of a commonly used vector template. These primers are used at a universal annealing temperature to amplify ten DNA fragments of accurate size ranging from 100bp to 1000bp. Our PCR-based method can provide size standards for an endless usage.

BACKGROUND: Human leukocyte antigen (HLA) is comprised of a highly polymorphic set of genes which determines the histocompatibility of organ transplantation. The present study was undertaken to identify HLA class I and class II allele, genotype and haplotype frequencies in renal transplant recipients and donors from West Central India. MATERIALS AND METHODS: HLA typing was carried out using Polymerase Chain Reaction-Sequence Specific Primer in 552 live related and unrelated renal transplant recipients and donors. RESULTS: The most frequent HLA class I and class II alleles and their frequencies in recipients were HLA-AFNx0101 (0.1685) and AFNx0102 (0.1649), HLA-BFNx0135 (0.1322), and HLA-DR beta 1 (DRB 1)FNx0115 (0.2192), whereas in donors, these were HLA-AFNx0102 (0.1848) and AFNx0101 (0.1667), HLA-BFNx0135 (0.1359), and HLA-DRB1FNx0115 (0.2409). The two-locus haplotype statistical analysis revealed HLA-AFNx0102-B61 as the most common haplotype with the frequency of 0.0487 and 0.0510 in recipients and donors, respectively. Further, among the three locus haplotypes HLA-AFNx0133-BFNx0144-DRB1FNx0107 and HLA-AFNx0102-BFNx0161-DRB1FNx0115 were the most common haplotypes with frequencies 0.0362 and 0.0326, respectively in recipients and 0.0236 and 0.0323, respectively in donors. Genotype frequency revealed a high prevalence of genotype HLA-AFNx0102/AFNx0124 in recipients (0.058) compared to donors (0.0109) whereas low prevalence of HLA-AFNx0101/AFNx0102 in recipients (0.0435) than in donors (0.0797). The phylogenetic and principal component analysis of HLA allele and haplotype frequency distribution revealed genetic similarities of various ethnic groups. Further, case control analysis provides preliminary evidence of association of HLA-A genotype (P < 0.05) with renal failure. CONCLUSION: This study will be helpful in suitable donor search besides providing valuable information for population genetics and HLA disease association analysis.

Sir, We read the article “Prevalence of lower urinary tract infection in South Indian type 2 diabetic subjects” by Janifer et al. with great interest. Diabetic patients with urinary tract infection is a common scenario in the nephrourology outpatient department. Treating them is a real challenge.[1] Here, we would like to highlight few important points. Firstly, fungal urinary tract infection in diabetic patients might be asymptomatic and should be sought in every diabetic patient with urinary tract infection.[2] Secondly, we feel that ultrasound-guided suprapubic aspiration of urine is useful in those patients with repeated contamination in urine culture report.[3] Thirdly, we believe that a minimum of 2 weeks of antibiotic therapy is necessary as eradication of the organism in diabetic patients is difficult. Improper duration of antibiotic will encourage resistance.[4] Fourthly, circumcision might be beneficial, especially in those patients who cannot maintain personal hygiene. We would like to know the author’s views regarding these points.

Cancer is one of the leading causes of death in most parts of the world and is a very serious cause of concern particularly in developing countries. In this work, we prepared and evaluated the aptamer-labeled paclitaxel-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles (Apt-PTX-PLGA NPs) which can ameliorate drug bioavailability and enable accurate drug targeting to cancer cells with controlled drug release for cancer therapy. Paclitaxel-loaded PLGA nanoparticles (PTX-PLGA NPs) were formulated by a single-emulsion/solvent evaporation method and were further surface-functionalized with a chemical cross-linker bis(sulfosuccinimidyl) suberate (BS3) to enable binding of aptamer on to the surface of the nanoparticles. The prepared nanoparticles were characterized by atomic force microscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. Cytotoxicity studies were carried out using normal human mammary epithelial cells (HMEC cells) and human glial cancer cells (GI-1 cells) by methylthiazolyldiphenyl-tetrazolium bromide assay and Alamar blue assay, which confirmed that PTX-PLGA NPs with aptamer conjugation (Apt-PTX-PLGA NPs) were comparatively non-toxic to HMEC cells while toxic to GI-1 cancer cells. Cellular uptake of PTX-PLGA NPs with and without aptamer conjugation was studied using GI-1 cells and monitored by confocal microscopy and phase contrast microscopy. Our studies demonstrated significant internalization and retention of nanoparticles inside the cells, inducing apoptosis. The preferential accumulation of PTX-PLGA NPs within the cancer cells were also confirmed by flow cytometry-based uptake studies. The results indicated that Apt-PTX-PLGA NPs could be a promising targeted therapeutic delivery vehicle for cancer treatment.