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Find video protocols related to scientific articles indexed in Pubmed.
Stability indicating studies on NMITLI 118RT+ (standardized extract of withania somnifera dunal).
Pharmacogn Mag
PUBLISHED: 07-24-2014
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Withania somnifera Dunal (Ashwagandha) is an Indian medicinal plant of great medicinal value; used in many clinically proven conditions. NMITLI-118RT+ is a candidate drug under a Council of Scientific and Industrial Research (CSIR) networking project. It is a chemotype of W. somnifera's root extract, which has been used for the present study.
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Massive periocular squamous cell carcinoma engulfing the globe: a rare case report.
Case Rep Oncol Med
PUBLISHED: 07-24-2014
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The eyelid tumors are rare neoplasms in head and neck oncological practice. The maximum incidence is after sixty years and most of the tumors are situated in the lower eyelid and medial canthus. A wide range of clinical presentations contribute to a high risk of misdiagnosis. The factors such as very large lesions, incomplete excision, histopathologic features such as poor differentiation, multicentric origin, pagetoid spread, and delayed diagnosis are associated with poor prognosis. Because of different tissues at eyelid level, a variety of tumor types and subtypes can arise, but most of them are carcinomas. A rare case of eyelid carcinoma spreading and engulfing the whole of globe which was treated by orbital exenteration and postoperative radiotherapy is presented with a disease-free follow-up of 10 months which, considering its size, is extremely rare. The early diagnosis and proper treatment of such rare tumors still remain the mainstay to predict favourable prognosis.
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Modified stethoscope for auscultation of temporomandibular joint sounds.
J Int Oral Health
PUBLISHED: 04-26-2014
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Purpose of this study was to modify the stethoscope which can auscultate the temporomandibular joint (TMJ) sounds more precisely than conventional stethoscope, and fabrication of stethoscope compatible software which analyses the auscultated sound and gives documentary evidence of that analysis in the form of graph.
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Molecular cloning and biochemical characterization of a recombinant sterol 3-O-glucosyltransferase from Gymnema sylvestre R.Br. catalyzing biosynthesis of steryl glucosides.
Biomed Res Int
PUBLISHED: 02-26-2014
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Gymnema sylvestre R.Br., a pharmacologically important herb vernacularly called Gur-Mar (sugar eliminator), is widely known for its antidiabetic action. This property of the herb has been attributed to the presence of bioactive triterpene glycosides. Although some information regarding pharmacology and phytochemical profiles of the plant are available, no attempts have been made so far to decipher the biosynthetic pathway and key enzymes involved in biosynthesis of steryl glucosides. The present report deals with the identification and catalytic characterization of a glucosyltransferase, catalyzing biosynthesis of steryl glycosides. The full length cDNA (2572 bp) contained an open reading frame of 2106 nucleotides that encoded a 701 amino acid protein, falling into GT-B subfamily of glycosyltransferases. The GsSGT was expressed in Escherichia coli and biochemical characterization of the recombinant enzyme suggested its key role in the biosynthesis of steryl glucosides with catalytic preference for C-3 hydroxyl group of sterols. To our knowledge, this pertains to be the first report on cloning and biochemical characterization of a sterol metabolism gene from G. sylvestre R.Br. catalyzing glucosylation of a variety of sterols of biological origin from diverse organisms such as bacteria, fungi, and plants.
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Expression and activity of Rac1 is negatively affected in the dehydroepiandrosterone induced polycystic ovary of mouse.
J Ovarian Res
PUBLISHED: 02-22-2014
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Polycystic ovarian syndrome (PCOS) is characterized by the presence of multiple follicular cysts, giving rise to infertility due to anovulation. This syndrome affects about 10% of women, worldwide. The exact molecular mechanism leading to PCOS remains obscure. RhoGTPase has been associated with oogenesis, but its role in PCOS remains unexplored. Therefore, we attempted to elucidate the Vav-Rac1 signaling in PCOS mice model.
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Comparative Growth Characteristics and Yield Attributes of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Higher Basidiomycetes) on Different Substrates in India.
Int J Med Mushrooms
PUBLISHED: 11-26-2013
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The present study investigated the effects of four forestry byproducts (sawdust of oak, mango, khair, and tuni) and three agricultural residues (paddy straw, wheat straw, and soybean waste) along with four supplements (wheat bran, rice bran, corn flour, and gram powder) on growth characteristics (spawn run and primordial formation) and yield of Ganoderma lucidum. There were significant differences (P=0.05) in yield regardless of substrates and supplements used in experimentation. Among substrates, agriculture residues supported better yield and biological efficiency of G. lucidum compared to forestry byproducts irrespective of the supplements. The highest yield (82.5 g) and biological efficiency (27.5%) were recorded from paddy straw supplemented with wheat bran, which invariably resulted in significantly higher yield compared to the unsupplemented check(s) or other supplements used in this study.
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ANN-QSAR model for virtual screening of androstenedione C-skeleton containing phytomolecules and analogues for cytotoxic activity against human breast cancer cell line MCF-7.
Comb. Chem. High Throughput Screen.
PUBLISHED: 11-19-2013
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The present study deals with the development of an artificial neural network based quantitative structure activity relationship (QSAR) model for virtual screening of active compounds which contain androstenedione carbonskeleton or their similar skeleton at the core. An empirical data modeling (with fitted data mapping) has been performed on the basis of bioassay record for human breast cancer cell line MCF7. The whole experimental data set was considered as test set. Standard feed-forward back-propagation neural network technique was applied to build the model. Leave-One- Out (LOO) cross-validation was performed to evaluate the performance of the model. The mapped model became the basis for selection best mapped compounds followed by development of Pharmacophore specific secondary QSAR model. In the present study, two best mapped molecules 4beta-hydroxy Withanolide-E and 7, 8-Dehydrocalotropin were used for development of the secondary QSAR model. These secondary-QSAR models were resulted with R2 LOOCV value 0.9845 and 0.9666 respectively. Docking studies, in silico phamacokinetic and toxicity analysis was also done for selected compounds. The screened compounds CID_73621, CID_16757497, CID_301751, CID_390666 and CID_46830222 were found with promising binding affinity value with aromatase with reference to the co-crystallized control compound androstenedione. Due to excellent extent of variance coverage in ANN based QSAR map model, it can be used as a robust non-linear QSAR model for androstenedione carbon-skeleton containing molecules and the protocol can be used to derive secondary QSAR models for other compounds set.
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L712V mutation in the androgen receptor gene causes complete androgen insensitivity syndrome due to severe loss of androgen function.
Steroids
PUBLISHED: 07-24-2013
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Inability to respond to the circulating androgens is named as androgen insensitivity syndrome (AIS). Mutations in the androgen receptor (AR) gene are the most common cause of AIS. A cause and effect relationship between some of these mutations and the AIS phenotype has been proven by in vitro studies. Several other mutations have been identified, but need to be functionally validated for pathogenicity. Screening of the AR mutations upon presumptive diagnosis of AIS is recommended. We analyzed a case of complete androgen insensitivity syndrome (CAIS) for mutations in the AR gene. Sequencing of the entire coding region revealed C>G mutation (CTT-GTT) at codon 712 (position according to the NCBI database) in exon 4 of the gene, resulting in replacement of leucine with valine in the ligand-binding domain of the AR protein. No incidence of this mutation was observed in 230 normal male individuals analyzed for comparison. In vitro androgen binding and transactivation assays using mutant clone showed approximately 71% loss of ligand binding and about 76% loss of transactivation function. We conclude that CAIS in this individual was due to L712V substitution in the androgen receptor protein.
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Platform switching: a narrative review.
Implant Dent
PUBLISHED: 07-12-2013
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Bone loss around the implant is 1 of the important factors affecting its success. Fitting an abutment of smaller circumference in comparison with the implant is known as platform switching. The concept gained importance as investigations found reduced crestal bone loss around such implants. Several studies have been conducted to understand its efficacy, mechanism of action, and the extent of switching that would provide best results.
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Resection and Reconstruction of Maxillary Class IIIc Defect in a Case of Adenoid Cystic Carcinoma: Cost-Sensitive Technique without Microvascular Grafts.
Case Rep Dent
PUBLISHED: 06-30-2013
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ACC is a rare malignant tumor that affects most commonly the major and minor salivary glands and rarely the paranasal sinuses, lacrimal gland, larynx, ear, vulva, and so forth. The maxillary sinus when affected is considered having a poor prognosis due to delayed diagnosis and delayed treatment credited to its slow spread, late symptoms, and complex anatomy which hampers surgical resection. The expressions of tumor markers too have a significant role in determining the prognosis. The treatment of choice consists of wide radical resection of the tumor followed by radiotherapy. Rehabilitation options in cases with huge maxillary defects still need further exploration.
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SRD5A2 gene polymorphisms affect the risk of breast cancer.
Breast
PUBLISHED: 06-07-2013
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Androgens in breast cancer have been studied alone and in correlation with estrogens as estrogen to testosterone ratio. 5-?-reductase is one of the important enzymes participating in androgen metabolism, which affects androgen activity by affecting conversion of testosterone to dihydrotestosterone. We hypothesized that polymorphisms in the SRD5A2 gene (encoding 5-?-reductase) may affect breast cancer risk by affecting total androgen activity. Complete coding region of the SRD5A2 gene was sequenced in a group of 628 patients and 244 control samples from three southern states (Tamil Nadu, Andhra Pradesh, and Karnataka) of India. We observed three common polymorphisms in this gene; namely, A49T, V89L, and (TA)n repeats. A49T locus was monomorphic in the study population, but V89L showed a strong correlation with breast cancer (P = 0.03, OR = 1.40, CI = 1.02-1.91). (TA)0/(TA)9 and (TA)9/(TA)9 genotypes were at a lower risk of breast cancer (P = 0.01, OR = 0.64, CI = 0.46-0.90). We conclude that SRD5A2 genotypes significantly affect breast cancer risk in the South Indian populations.
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Non-Hodgkins lymphoma of maxillary sinus: An unusual presentation.
Ann Maxillofac Surg
PUBLISHED: 05-11-2013
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Non-Hodgkins Lymphoma (NHL) are a group of neoplasms that originate from the cells of the lymphoreticular system. Forty percent of Non-Hodgkins lymphoma arises from extranodal sites. The nasal cavities and paranasal sinuses are rarely affected by primary NHL. Common primary extranodal sites of lymphomas include stomach, liver, soft tissue, dura, bone, intestine and bone marrow. Most patients present with rapidly enlarging masses, often with symptoms both locally and systemically (fever, recurrent night sweats, or weight loss). The vast majority of patients with localized disease are curable with combined modality therapy or combination chemotherapy alone. About 50% patients are cured with doxorubicin based combination chemotherapy and rituximab. An atypical case of extranodal Non-Hodgkins lymphoma of maxillary sinus is discussed.
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A mitochondrial DNA variant 10398G>A in breast cancer among South Indians: an original study with meta-analysis.
Mitochondrion
PUBLISHED: 05-09-2013
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The m.10398G>A polymorphism in the MT-ND3 gene has been linked to the manifestation of several neurodegenerative disorders and cancers. Several research groups have analyzed the association between m.10398G>A polymorphism and breast cancer; however, the results do not follow a consensus. We have studied this polymorphism in three Dravidian populations from South India. Analysis on 716 cases and 724 controls found no association between m.10398G>A polymorphism and breast cancer [OR = 0.916 (0.743-1.128); P = 0.409]. Menopausal stratification also revealed no significant association in either pre-menopausal or post-menopausal breast cancer groups. In addition, we undertook a meta-analysis on 16 study groups, comprising a total of 7202 cases and 7490 controls. The pooled odds ratio suggested no significant association of m.10398G>A substitution with breast cancer [OR = 1.016 (0.85-1.22); P = 0.86]. In conclusion, there is no evidence of association between m.10398G>A polymorphism and breast cancer risk among South Indian women. Meta-analysis suggested no overall correlation between this polymorphism and breast cancer risk.
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Purification and physicokinetic characterization of a gluconolactone inhibition-insensitive ?-glucosidase from Andrographis paniculata nees. Leaf.
Prep. Biochem. Biotechnol.
PUBLISHED: 04-16-2013
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A gluconolactone inhibition-insensitive ?-glucosidase from Andrographis paniculata (Acanthaceae) leaves has been isolated, homogeneity purified, and characterized for its physicokinetic properties. The purified enzyme appeared to be a monomeric structure with native molecular weight about 60 kD. The enzyme exhibited optimum pH 5.5 and pI 4.0, meso-thermostability and high temperature optimum (55°C) for catalytic activity, with activation energy of 6.8 kcal Mol(-1). The substrate saturation kinetics studies of the enzyme revealed a Michaelis-Menten constant (Km) of 0.25 mM for pNPG and catalytic efficiency (Kcat/Km) of 52,400 M (-1) s(-1), respectively. Substrate specificity of the enzyme was restricted to ?-linked gluco-, manno- and fuco-conjugates. The gluconolactone inhibition insensitivity was evident from its very low inhibition at millimolar inhibitor concentrations. Interestingly, the enzyme showed geraniol transglucosylating activity with pNPG as glucosyl donor but not with cellobiose. The catalytic activity of the enzyme has been reported to be novel with respect to its activity and preferences from a medicinal plant resource.
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Activated charcoal-mediated RNA extraction method for Azadirachta indica and plants highly rich in polyphenolics, polysaccharides and other complex secondary compounds.
BMC Res Notes
PUBLISHED: 03-20-2013
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High quality RNA is a primary requisite for numerous molecular biological applications but is difficult to isolate from several plants rich in polysaccharides, polyphenolics and other secondary metabolites. These compounds either bind with nucleic acids or often co-precipitate at the final step and many times cannot be removed by conventional methods and kits. Addition of vinyl-pyrollidone polymers in extraction buffer efficiently removes polyphenolics to some extent, but, it failed in case of Azadirachta indica and several other medicinal and aromatic plants.
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Significance of obesity markers and adipocytokines in high grade and high stage prostate cancer in North Indian men - a cross-sectional study.
Cytokine
PUBLISHED: 03-15-2013
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Prostate cancer (CaP) in India is the 10th most common malignancy affecting men. CaP incidence in India is low, but rising like other countries. The reasons for this racial disparity are uncertain. The foremost reasons that may underlie regional/ethnic differences are genetic polymorphisms, altered hormonal status, socioeconomic status, and obesity. This study aimed at investigating the role of adipocytokines in stimulating the promotion and progression of CaP.
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Association of maternal and fetal MTHFR A1298C polymorphism with the risk of pregnancy loss: a study of an Indian population and a meta-analysis.
Fertil. Steril.
PUBLISHED: 01-26-2013
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To study the genetic association between methylenetetrahydrofolate reductase (MTHFR) A1298 polymorphism and recurrent pregnancy loss (RPL).
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Mucuna pruriens and its major constituent L-DOPA recover spermatogenic loss by combating ROS, loss of mitochondrial membrane potential and apoptosis.
PLoS ONE
PUBLISHED: 01-22-2013
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The Ayurvedic medicinal system claims Mucuna pruriens (MP) to possess pro-male fertility, aphrodisiac and adaptogenic properties. Some scientific evidence also supports its pro-male fertility properties; however, the mechanism of its action is not yet clear. The present study aimed at demonstrating spermatogenic restorative efficacy of MP and its major constituent L-DOPA (LD), and finding the possible mechanism of action thereof in a rat model.
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A rare non-synonymous c.102C>G SNP in the IFNB1 gene might be a risk factor for cerebral malaria in Indian populations.
Infect. Genet. Evol.
PUBLISHED: 01-17-2013
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Interferon beta1 (IFNB1) is a type I interferon that is mainly known for its antiviral activity, but it also regulates a number of anti-inflammatory and immunomodulatory functions. Studies on mouse models of cerebral malaria have established that IFNB1 regulates severe malaria pathogenesis and increases overall survival against malaria. It down-regulates pro-inflammatory cytokines: TNF, IFNG and ICAM-1, resulting in decreased adherence of Plasmodium falciparum parasitized RBC to capillary wall, entry into the brain and delayed onset of death. Therefore, we hypothesized that variations in IFNB1 gene could regulate malarial pathogenesis. We re-sequenced the complete IFNB1 gene along with 900bp of 5 up-stream and 500bp of 3-UTR in 437 individuals from malaria endemic regions of the Orissa and Chhattisgarh states of India. The subjects comprised of 173 cases of severe malaria, 101 of mild malaria, and 156 ethnically matched asymptomatic controls. Data were statistically compared between cases and controls for their possible association with P. falciparum malarial outcome. Two single nucleotide polymorphisms (SNPs): a synonymous c.153C>T (rs1051922) and a non-synonymous substitution c.102C>G (rs139262191, p.Ser34Arg) were identified. The genotype and allele distribution of c.153C>T did not differ significantly between the study groups [mild, ?(2)2=4.10, p-value<0.13 and severe ?(2)2=0.06, p-value<0.97]. Interestingly, the rare non-synonymous SNP (rs139262191) was observed only in malaria patients. The differences between all cases and controls did not reach statistical significance, however, a statistically significant difference was observed between the asymptomatic control group and the cerebral malaria group [OR=20.32, 95% CI=1.08-382.63, p-value=0.044]. Moreover, the genotypes between cerebral malaria positive and negative groups were not significantly different [OR=5.58, 95% CI=0.61-50.97, p-value=0.123]. Our findings suggest that the IFNB1 variant, p.Ser34Arg, might be a risk factor for cerebral malaria in Indian populations.
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Androgen insensitivity syndrome: ten years of our experience.
Front Biosci (Elite Ed)
PUBLISHED: 01-02-2013
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Abnormalities of secondary sexual differentiation manifest in varying degrees depending upon the severity of the underlying cause. Primary amenorrhea in phenotypic females is caused by several different factors, including hormonal imbalance, nutritional deficiency and sex differentiation abnormalities. Androgen insensitivity syndrome (AIS) accounts for a large proportion of such cases in phenotypic females but genetically male individuals. Over the past 10 years, we have collected data related to androgen insensitivity from more than 150 cases. The research identified several important but neglected facts about this syndrome; including the identification of mutations in 39% of the cases and the establishment of the cause of pathogenesis in 60% of them. The most intriguing facts were uncovered in relation to late presentation of the AIS cases, little awareness among patients and family members, no consensus on the age of performing gonadectomy, and reluctance of the patients to undergo recommended surgery. These issues need immediate attention to improve healthcare and management of AIS cases. This article summarizes our observations about AIS with an aim to spread awareness among patients and clinicians.
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Strong Impact of TGF-?1 Gene Polymorphisms on Breast Cancer Risk in Indian Women: A Case-Control and Population-Based Study.
PLoS ONE
PUBLISHED: 01-01-2013
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TGF-?1 is a multi-functional cytokine that plays an important role in breast carcinogenesis. Critical role of TGF-?1 signaling in breast cancer progression is well documented. Some TGF-?1 polymorphisms influence its expression; however, their impact on breast cancer risk is not clear.
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Androgen receptor CAG repeats length polymorphism and the risk of polycystic ovarian syndrome (PCOS).
PLoS ONE
PUBLISHED: 01-01-2013
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Polycystic ovarian syndrome (PCOS) refers to an inheritable androgen excess disorder characterized by multiple small follicles located at the ovarian periphery. Hyperandrogenism in PCOS, and inverse correlation between androgen receptor (AR) CAG numbers and AR function, led us to hypothesize that CAG length variations may affect PCOS risk.
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Tropine Forming Tropinone Reductase Gene from Withania somnifera (Ashwagandha): Biochemical Characteristics of the Recombinant Enzyme and Novel Physiological Overtones of Tissue-Wide Gene Expression Patterns.
PLoS ONE
PUBLISHED: 01-01-2013
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Withania somnifera is one of the most reputed medicinal plants of Indian systems of medicine synthesizing diverse types of secondary metabolites such as withanolides, alkaloids, withanamides etc. Present study comprises cloning and E. coli over-expression of a tropinone reductase gene (WsTR-I) from W. somnifera, and elucidation of biochemical characteristics and physiological role of tropinone reductase enzyme in tropane alkaloid biosynthesis in aerial tissues of the plant. The recombinant enzyme was demonstrated to catalyze NADPH-dependent tropinone to tropine conversion step in tropane metabolism, through TLC, GC and GC-MS-MS analyses of the reaction product. The functionally active homodimeric ?60 kDa enzyme catalyzed the reaction in reversible manner at optimum pH 6.7. Catalytic kinetics of the enzyme favoured its forward reaction (tropine formation). Comparative 3-D models of landscape of the enzyme active site contours and tropinone binding site were also developed. Tissue-wide and ontogenic stage-wise assessment of WsTR-I transcript levels revealed constitutive expression of the gene with relatively lower abundance in berries and young leaves. The tissue profiles of WsTR-I expression matched those of tropine levels. The data suggest that, in W. somnifera, aerial tissues as well possess tropane alkaloid biosynthetic competence. In vivo feeding of U-[(14)C]-sucrose to orphan shoot (twigs) and [(14)C]-chasing revealed substantial radiolabel incorporation in tropinone and tropine, confirming the de novo synthesizing ability of the aerial tissues. This inherent independent ability heralds a conceptual novelty in the backdrop of classical view that these tissues acquire the alkaloids through transportation from roots rather than synthesis. The TR-I gene expression was found to be up-regulated on exposure to signal molecules (methyl jasmonate and salicylic acid) and on mechanical injury. The enzymes catalytic and structural properties as well as gene expression profiles are discussed with respect to their physiological overtones.
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Significant impact of the MTHFR polymorphisms and haplotypes on male infertility risk.
PLoS ONE
PUBLISHED: 01-01-2013
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Methylenetetrahydrofolate reductase (MTHFR) converts 5,10-methylene tetrahydrofolate to 5-methyl tetrahydrofolate and affects the activity of cellular cycles participating in nucleotide synthesis, DNA repair, genome stability, maintenance of methyl pool, and gene regulation. Genetically compromised MTHFR activity has been suggested to affect male fertility. The objective of the present study was to find the impact on infertility risk of c.203G>A, c.1298A>C, and c.1793G>A polymorphisms in the MTHFR gene.
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De novo assembly, functional annotation and comparative analysis of Withania somnifera leaf and root transcriptomes to identify putative genes involved in the withanolides biosynthesis.
PLoS ONE
PUBLISHED: 01-01-2013
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Withania somnifera is one of the most valuable medicinal plants used in Ayurvedic and other indigenous medicine systems due to bioactive molecules known as withanolides. As genomic information regarding this plant is very limited, little information is available about biosynthesis of withanolides. To facilitate the basic understanding about the withanolide biosynthesis pathways, we performed transcriptome sequencing for Withania leaf (101L) and root (101R) which specifically synthesize withaferin A and withanolide A, respectively. Pyrosequencing yielded 8,34,068 and 7,21,755 reads which got assembled into 89,548 and 1,14,814 unique sequences from 101L and 101R, respectively. A total of 47,885 (101L) and 54,123 (101R) could be annotated using TAIR10, NR, tomato and potato databases. Gene Ontology and KEGG analyses provided a detailed view of all the enzymes involved in withanolide backbone synthesis. Our analysis identified members of cytochrome P450, glycosyltransferase and methyltransferase gene families with unique presence or differential expression in leaf and root and might be involved in synthesis of tissue-specific withanolides. We also detected simple sequence repeats (SSRs) in transcriptome data for use in future genetic studies. Comprehensive sequence resource developed for Withania, in this study, will help to elucidate biosynthetic pathway for tissue-specific synthesis of secondary plant products in non-model plant organisms as well as will be helpful in developing strategies for enhanced biosynthesis of withanolides through biotechnological approaches.
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Laparoscopic female sterilisation by a single port through monitor--a better alternative.
J Indian Med Assoc
PUBLISHED: 12-23-2011
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Female sterilisation by tubal occlusion method by laparocator is most widely used and accepted technique of all family planning measures all over the world. After the development of laparoscopic surgery in all faculties of surgery by monitor, now laparoscopic female sterilisation has been developed to do under monitor control by two ports--one for laparoscope and second for ring applicator. But the technique has been modified using single port with monitor through laparocator in which camera is fitted on the eye piece of laparocator (the same laparocator which is commonly used in camps without monitor since a long time in India). In this study over a period of about 2 years, a total 2011 cases were operated upon. In this study, I used camera and monitor through a single port by laparocator to visualise as well as to apply ring on fallopian tubes. The result is excellent and is a better alternative to conventional laparoscopic sterilisation and double puncture technique through camera--which give two scars and an extra assistant is required. However, there was no failure and the strain on surgeons eye was minimum. Single port is much easier, safe, equally effective and better acceptable method.
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Mining and survey of simple sequence repeats in wheat rust Puccinia sp.
Bioinformation
PUBLISHED: 11-02-2011
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The abundance and inherent potential for extensive allelic variations in simple sequence repeats (SSRs) or microsatellites resulted in valuable source for genetic markers in eukaryotes. In this study, we analyzed and compared the abundance and organisation of SSR in the genome of two important fungal pathogens of wheat, brown or leaf rust (Puccinia triticina) and black or stem rust (Puccinia graminis f. sp. tritici). P. triticina genome with two fold genome size as compared to P. graminis tritici has lower relative abundance and SSR density. The distribution pattern of different SSR motifs provides the evidence of greater accumulation of dinucleotide followed by trinucleotide repeats. More than two-hundred different types of repeat motifs were observed in the genomes. The longest SSR motifs varied in both genomes and some of the repeat motifs are found in higher frequency. The information about survey of relative abundance, relative density, length and frequency of different repeat motifs in Puccinia sp. will be useful for developing SSR markers that could find several applications in analysis of fungal genome such as genetic diversity, population genetics, race identification and acquisition of new virulence.
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Secular changes in the semen quality in India during the past 33 years.
J. Androl.
PUBLISHED: 10-20-2011
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Several studies have described a change in semen quality over last 60 years in many regions of the world, but such studies are lacking for the Indian population. In the present study, we aimed at investigating if semen quality has changed during the last 3 decades in normal healthy Indians. We retrieved data on semen quality for Indian men without a history of infertility from published studies and analyzed it for trends in individual semen quality parameters using regression modeling. The semen parameters of 19,734 normal, healthy men from studies published over the past 33 years (from 1978 onward) were used for this purpose. Linear regression analysis weighted by sample size and controlling for age of the subjects revealed a significant decline in sperm motility and a significant increase in sperm concentration. Other semen parameters did not change significantly over this period. In conclusion, there has been a genuine decline in sperm motility and an increase in sperm concentration, which is corroborated by a recent study on Indian subjects.
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Thyroid, spermatogenesis, and male infertility.
Front Biosci (Elite Ed)
PUBLISHED: 05-31-2011
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Since the identification of thyroid hormone receptors on the testes, thyroid has been suggested to have a significant impact on the male reproductive tract, spermatogenesis, and male fertility. Several research articles on the role of thyroid in spermatogenesis or male infertility have been published in the last three decades. We conducted an exhaustive literature search was conducted in order to create an up-to-date review of literature. This review aims to discuss the impact of thyroid on testicular development, spermatogenesis, hypo- or hyper- thyroidism and male infertility, and the management of thyroid related abnormal semen profile. The literature revealed that thyroid significantly impacts testicular development and that abnormal thyroid profile affects semen quality and male fertility by compromising testicular size, sperm motility and ejaculate volume. A clear link exists between thyroid hormones, testicular development and spermatogenesis. Thyroid disease negatively affects spermatogenesis and consequently may cause male infertility. In such cases, infertility is reversible, but more studies need to be conducted, especially in post-pubertal males to cement the current findings.
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Role of genetic polymorphisms of CYP1A1, CYP3A5, CYP2C9, CYP2D6, and PON1 in the modulation of DNA damage in workers occupationally exposed to organophosphate pesticides.
Toxicol. Appl. Pharmacol.
PUBLISHED: 05-10-2011
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Organophosphate pesticides (OPs) are primarily metabolized by several xenobiotic metabolizing enzymes (XMEs). Very few studies have explored genetic polymorphisms of XMEs and their association with DNA damage in pesticide-exposed workers. The present study was designed to determine the role of genetic polymorphisms of CYP1A1, CYP3A5, CYP2C9, CYP2D6, and PON1 in the modulation of DNA damage in workers occupationally exposed to OPs. We examined 284 subjects including 150 workers occupationally exposed to OPs and 134 normal healthy controls. The DNA damage was evaluated using the alkaline comet assay and genotyping was done using PCR-RFLP. The results revealed that the PONase activity toward paraoxonase and AChE activity was found significantly lowered in workers as compared to control subjects (p<0.001). Workers showed significantly higher DNA damage compared to control subjects (14.37±2.15 vs. 6.24±1.37 tail% DNA, p<0.001). Further, the workers with CYP2D6*3PM and PON1 (QQ and MM) genotypes were found to have significantly higher DNA damage when compared to other genotypes (p<0.05). In addition, significant increase in DNA damage was also observed in workers with concomitant presence of certain CYP2D6 and PON1 (Q192R and L55M) genotypes which need further extensive studies. In conclusion, the results indicate that the PON1 and CYP2D6 genotypes can modulate DNA damage elicited by some OPs possibly through gene-environment interactions.
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Influence of CYP2C9, GSTM1, GSTT1 and NAT2 genetic polymorphisms on DNA damage in workers occupationally exposed to organophosphate pesticides.
Mutat. Res.
PUBLISHED: 05-03-2011
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Previous studies have revealed that organophosphate pesticides (OPs) are primarily metabolized by xenobiotic metabolizing enzymes (XMEs). Very few studies have explored genetic polymorphisms of XMEs and their association with DNA damage in pesticides-exposed workers. Present study was designed to determine the influence of CYP2C9, GSTM1, GSTT1 and NAT2 genetic polymorphisms on DNA damage in workers occupationally exposed to OPs. We examined 268 subjects including 134 workers occupationally exposed to OPs and an equal number of normal healthy controls. The DNA damage was evaluated using alkaline comet assay and genotyping was done using individual polymerase chain reaction (PCR) or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Acetylcholinesterase and paraoxonase activity were found to be significantly lowered in workers as compared to control subjects which were analyzed as biomarkers of toxicity due to OPs exposure (p<0.001). Workers showed significantly higher DNA tail moment (TM) compared to control subjects (14.32±2.17 vs. 6.24±1.37 tail % DNA, p<0.001). GSTM1 null genotype was found to influence DNA TM in workers (p<0.05). DNA TM was also found to be increased with concomitant presence of NAT2 slow acetylation and CYP2C9*3/*3 or GSTM1 null genotypes (p<0.05). DNA TM was found increased in NAT2 slow acetylators with mild and heavy smoking habits in control subjects and workers, respectively (p<0.05). The results of this study suggest that GSTM1 null genotypes, and an association of NAT2 slow acetylation genotypes with CYP2C9*3/*3 or GSTM1 null genotypes may modulate DNA damage in workers occupationally exposed to OPs.
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Strong association of 677 C>T substitution in the MTHFR gene with male infertility--a study on an indian population and a meta-analysis.
PLoS ONE
PUBLISHED: 04-23-2011
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Methylenetetrahydrofolate reductase (MTHFR) is an important enzyme of folate and methionine metabolism, making it crucial for DNA synthesis and methylation. The objective of this study was to analyze MTHFR gene 677C>T polymorphism in infertile male individuals from North India, followed by a meta-analysis on our data and published studies.
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Epigenetics, spermatogenesis and male infertility.
Mutat. Res.
PUBLISHED: 04-07-2011
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Epigenetic modifications characterized by DNA methylation, histone modifications, and chromatin remodeling are important regulators in a number of biological processes, including spermatogenesis. Several genes in the testes are regulated through epigenetic mechanisms, indicating a direct influence of epigenetic mechanisms on the process of spermatogenesis. In the present article, we have provided a comprehensive review of the epigenetic processes in the testes, correlation of epigenetic aberrations with male infertility, impact of environmental factors on the epigenome and male fertility, and significance of epigenetic changes/aberrations in assisted reproduction. The literature review suggested a significant impact of epigenetic aberrations (epimutations) on spermatogenesis, and this could lead to male infertility. Epimutations (often hypermethylation) in several genes, namely MTHFR, PAX8, NTF3, SFN, HRAS, JHM2DA, IGF2, H19, RASGRF1, GTL2, PLAG1, D1RAS3, MEST, KCNQ1, LIT1, and SNRPN, have been reported in association with poor semen parameters or male infertility. Environmental toxins/drugs may affect fertility via epigenetic modifications. For example, 5-aza-2-deoxycytidine, an anticancer agent, causes a decrease in global DNA methylation that leads to altered sperm morphology, decreased sperm motility, decreased fertilization capacity, and decreased embryo survival. Similarly, Endocrine disruptors, such as methoxychlor (an estrogenic pesticide) and vinclozolin (an anti-androgenic fungicide) have been found by experiments on animals to affect epigenetic modifications that may cause spermatogenic defects in subsequent generations. Assisted reproduction procedures that have been considered rather safe, are now being implicated in inducing epigenetic changes that could affect fertility in subsequent generations. Techniques such as intracytoplasmic sperm injection (ICSI) and round spermatid injection (ROSI) may increase the incidence of imprinting disorders and adversely affect embryonic development by using immature spermatozoa that may not have established proper imprints or global methylation. Epigenetic changes, in contrast to genetic aberrations, may be less deleterious because they are potentially reversible. Further research could identify certain drugs capable of reversing epigenetic changes.
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Modeling and phylogeny analysis of bread wheat MnSOD.
Bioinformation
PUBLISHED: 03-11-2011
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Superoxide dismutase (SOD) acts as first line of defense against oxidative and genetic stress. Manganese superoxide dismutase (MnSOD), found in mitochondria or peroxisomes, contains Mn(III) at the active site. Therefore, it is of interest to study MnSOD from bread wheat (a grain crop). However, a structure model is not yet solved for bread wheat MnSOD. Hence, we describe the structure model of bread wheat MnSOD developed using homology model. The model provides molecular insight to metal binding molecular function towards the understanding of oxidative stress resistance in plants. The distinction of bread wheat (a monocot) MnSOD from dicots is also shown using phylogenetic analysis.
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A novel Arg615Ser mutation of androgen receptor DNA-binding domain in three 46,XY sisters with complete androgen insensitivity syndrome and bilateral inguinal hernia.
Fertil. Steril.
PUBLISHED: 03-01-2011
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To present clinical, genetic, biochemical, and molecular findings in a family with three sisters with complete androgen insensitivity syndrome (CAIS) and bilateral inguinal hernia.
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Genetic polymorphisms of GSTM1, GSTT1 and GSTP1 and susceptibility to DNA damage in workers occupationally exposed to organophosphate pesticides.
Mutat. Res.
PUBLISHED: 02-07-2011
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GSTM1, T1 and P1 are important enzymes of glutathione S-transferases (GSTs), involved in the metabolism of many endogenous and exogenous compounds. Individual genetic variation in these metabolizing enzymes may influence the metabolism of their substrates. The present study was designed to determine the genotoxic effects using DNA damage and its association with GSTM1, GSTT1, and GSTP1 (Ile105Val) genetic polymorphisms in workers occupationally exposed to organophosphate pesticides (OPs). We examined 230 subjects including 115 workers occupationally exposed to OPs and an equal number of normal healthy controls. The DNA damage was evaluated using the alkaline comet assay and genotyping was done using individual PCR or PCR-RFLP. Significantly higher DNA tail moment (TM) was observed in workers as compared to control subjects (14.41 ± 2.25 vs. 6.36 ± 1.41 tail % DNA, p<0.001). The results revealed significantly higher DNA TM in workers with GSTM1 null genotype than those with GSTM1 positive (15.18 vs. 14.15 tail % DNA, p=0.03). A significantly higher DNA TM was also observed in workers with homozygous Ile-Ile GSTP1 genotype than heterozygous (Ile-Val) and mutant (Val-Val) GSTP1 genotype (p=0.02). In conclusion, the results show that null deletion of GSTM1 and homozygote wild GSTP1 genotype could be related to inter-individual differences in DNA damage arises from the gene-environment interactions in workers occupationally exposed to OPs.
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Insights of interaction between small and large subunits of ADP-glucose pyrophosphorylase from bread wheat (Triticum aestivum L.).
Bioinformation
PUBLISHED: 01-29-2011
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Lack of knowledge of three dimensional structures of small and large subunits of ADP- glucose pyrophosphorylase (AGPase) in wheat has hindered efforts to understand the binding specifities of substrate and catalytic mechanism. Thus, to understand the structure activity relationship, 3D structures were built by homology modelling based on crystal structure of potato tuber ADP-glucose pyrophosphorylase. Selected models were refined by energy minimization and further validated by Procheck and Prosa-web analysis. Ramachandran plot showed that overall main chain and side chain parameters are favourable. Moreover, Z-score of the models from Prosa-web analysis gave the conformation that they are in the range of the template. Interaction analysis depicts the involvement of six amino acids in hydrogen bonding (AGP-SThr422-AGP-LMet138, AGP- SArg420-AGP-LGly47, AGP-SSer259-AGP-LSer306, AGP-SGlu241-AGP-LIle311, AGPSGln113- AGP-LGlu286 and AGP-SGln70-AGP-LLys291). Fifteen amino acids of small subunit were able to make hydrophobic contacts with seventeen amino acids of large subunit. Furthermore, decrease in the solvent accessible surface area in the amino acids involved in interaction were also reported. All the distances were formed in between 2.27 to 3.78?. The present study focussed on heterodimeric structure of (AGPase). This predicted complex not only enhance our understanding of the interaction mechanism between these subunits (AGP-L and AGP-S) but also enable to further study to obtain better variants of this enzyme for the improvement of the plant yield.
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Evaluation of Volvariella volvacea Strains for Yield and Diseases/Insect-Pests Resistance Using Composted Substrate of Paddy Straw and Cotton Mill Wastes.
Indian J. Microbiol.
PUBLISHED: 01-25-2011
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Out of the 3 parent strains and 4 single spore isolates of Volvariella volvacea evaluated, strain, OE-274 gave earliest yield in 11.25-11.50 days post-spawning in all 4 trials. The yield varied in different strains in different trials and it was highest in strain, OE-272 in trial 1, SSI, OE-55-08 in trial 2, and strain, OE-274 in trial 3 and 4. In overall average, highest yield was in strain, OE-272, closely followed by strain, OE-274. The number of fruiting bodies per q substrate also varied in different strains in different trials. Highest numbers were in strain, OE-272, SSIs, OE-55-08 and OE-12-22, and strain, OE-210 in trial 1, 2, 3 and 4, respectively. Highest fruiting body wt was in strain, OE-274 in all 4 trials. The yield during different weeks of cropping varied in different strains but invariably it was highest in first week, which accounted for 60-70% of the total yield. The fruiting bodies of strain, OE-274 were of bigger size, brownish, toughest and with least tendency of veil opening, while that of strain, OE-272 and SSI, OE-55-08 were whitish to grayish-white, oblong, medium size, delicate and lesser tendency of veil opening. The strain, OE-274 and SSI, OE-55-08 exhibited higher resistance against the growth of competitor moulds and infestations of insect-pests, while strain, OE-272 exhibited highest susceptibility to insect-pests infestation.
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High fat diet prevents over-crowding induced decrease of sex ratio in mice.
PLoS ONE
PUBLISHED: 01-25-2011
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Adaptive theory predicts that mothers would be advantaged by adjusting the sex ratio of their offspring in relation to their offsprings future reproductive success. In the present study, we tested the effect of housing mice under crowded condition on the sex ratio and whether the fat content of the diet has any influence on the outcome of pregnancies. Three-week-old mice were placed on the control diet (NFD) for 3 weeks. Thereafter the mice were allotted randomly to two groups of 7 cages each with 4, 6, 8, 10, 12, 14, and 16 mice in every cage to create increasing crowding gradient and fed either NFD or high fat diet (HFD). After 4 weeks, dams were bred and outcomes of pregnancy were analyzed. The average dam body weight (DBW) at conception, litter size (LS) and SR were significantly higher in HFD fed dams. Further, male biased litters declined with increasing crowding in NFD group but not in HFD. The LS and SR in NFD declined significantly with increasing crowding, whereas only LS was reduced in HFD group. We conclude that female mice housed under overcrowding conditions shift offspring SR in favor of daughters in consistent with the TW hypothesis and high fat diet reduces this influence of overcrowding.
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Paraoxonase-1 genetic polymorphisms and susceptibility to DNA damage in workers occupationally exposed to organophosphate pesticides.
Toxicol. Appl. Pharmacol.
PUBLISHED: 01-24-2011
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Human paraoxonase 1 (PON1) is a lipoprotein-associated enzyme involved in the detoxification of organophosphate pesticides (OPs) by hydrolyzing the bioactive oxons. Polymorphisms of the PON1 gene are responsible for variation in the expression and catalytic activity of PON1 enzyme. In the present study, we have determined (a) the prevalence of two common PON1 polymorphisms, (b) the activity of PON1 and acetylcholinesterase enzymes, and (c) the influence of PON1 genotypes and phenotypes variation on DNA damage in workers exposed to OPs. We examined 230 subjects including 115 workers exposed to OPs and an equal number of normal healthy controls. The results revealed that PON1 activity toward paraoxon (179.19±39.36 vs. 241.52±42.32nmol/min/ml in controls) and phenylacetate (112.74±17.37 vs. 134.28±25.49?mol/min/ml in controls) was significantly lower in workers than in control subjects (p<0.001). No significant difference was observed in the distribution of genotypes and allelic frequencies of PON1(192)QR (Gln/Arg) and PON1(55)LM (Leu/Met) in workers and control subjects (p>0.05). The PON1 activity toward paraoxonase was found to be significantly higher in the R/R (Arg/Arg) genotypes than Q/R (Gln/Arg) and lowest in Q/Q (Gln/Gln) genotypes in both workers and control subjects (p<0.001). For PON1(55)LM (Leu/Met), PON1 activity toward paraoxonase was observed to be higher in individuals with L/L (Leu/Leu) genotypes and lowest in individuals with M/M (Met/Met) genotypes in both groups (p<0.001). No influence of PON1 genotypes and phenotypes was seen on the activity of acetylcholinesterase and arylesterase. The DNA damage was observed to be significantly higher in workers than in control subjects (p<0.05). Further, the individuals who showed least paraoxonase activity i.e., those with (Q/Q [Gln/Gln] and M/M [Met/Met]) genotypes showed significantly higher DNA damage compared to other isoforms in workers exposed to OPs (p<0.05). The results indicate that the individuals with PON1 Q/Q and M/M genotypes are more susceptible toward genotoxicity. In conclusion, the study suggests wide variation in enzyme activities and DNA damage due to polymorphisms in PON1 gene, which might have an important role in the identification of individual risk factors in workers occupationally exposed to OPs.
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Withania somnifera improves semen quality by combating oxidative stress and cell death and improving essential metal concentrations.
Reprod. Biomed. Online
PUBLISHED: 01-17-2011
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This study investigated the effect of a 3-month treatment with Withania somnifera on apoptosis and intracellular reactive oxygen species (ROS) concentration of spermatozoa and the metal ions copper, zinc, iron and gold in seminal plasma from infertile men (normozoospermic, n=25; oligozoospermic, n=25; and asthenozoospermic, n=25). The apoptotic and necrotic cell distribution were analysed by annexin-V binding and propidium iodide uptake using flow cytometry. ROS generation was measured by fluorescence intensity and metal ions were analysed by atomic absorption spectrophotometry. The results demonstrated that, prior to treatment, sperm apoptosis and intracellular ROS concentrations were significantly higher in all groups of infertile men compared with controls (P<0.01 to P<0.001). Similarly, the concentrations of the essential metal ions Cu(2+), Zn(2+), Fe(2+) and Au(2+) in seminal plasma were lower. Treatment with W. somnifera significantly reduced apoptosis in normozoospermic and oligozoospermic men and ROS concentrations in oligozoospermic and asthenozoospermic men (all P<0.05). Treatment also significantly improved metal ion concentrations in infertile men (P<0.01). It is concluded that W. somnifera improves semen quality by reducing oxidative stress and cell death, as well as improving essential metal ion concentrations. The aim of this study was to investigate the effect of 3-month treatment with Withania somnifera on apoptosis and intracellular reactive oxygen species (ROS) concentration in spermatozoa from infertile men. Before and following treatment, sperm apoptosis and concentrations of intracellular ROS and the metal ions copper, zinc, iron, and gold in seminal plasma were measured. The apoptotic and necrotic cell distribution were analysed by annexin-V binding and propidium iodide uptake using flow cytometry. ROS generation was measured by fluorescence intensity and metal ions were analysed by atomic absorption spectrophotometry. The results demonstrated that prior, to treatment, apoptosis and intracellular ROS concentrations were significantly higher in all groups of infertile men compared with controls. Similarly, the concentrations of the essential metal ions Cu(2+), Zn(2+), Fe(2+) and Au(2+) in seminal plasma were lower. Treatment with W. somnifera significantly reduced apoptosis and ROS concentrations and improved metal ion concentrations in infertile subjects. It is concluded that W. somnifera improves semen quality by reducing oxidative stress and cell death and improving essential metal ion concentrations.
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Analysis of simple sequence repeats (SSRs)dynamics in fungus Fusarium graminearum.
Bioinformation
PUBLISHED: 01-06-2011
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The abundance and inherent potential for variations in simple sequence repeats (SSRs) or microsatellites resulted in valuable source for genetic markers in eukaryotes. We describe the organization and abundance of SSRs in fungus Fusarium graminearum (causative agent for Fusarium head blight or head scab of wheat). We identified 1705 SSRs of various nucleotide repeat motifs in the sequence database of F. graminearum. It is observed that mononucleotide repeats (62%) were most abundant followed by di- (20%) and trinucleotide repeats (14%). It is noted that tetra-, penta- and hexanucleotide repeats accounted for only 4% of SSRs. The estimated frequency of Class I SSRs (perfect repeats ?20 nucleotides) was one SSR per 124.5 kb, whereas the frequency of Class II (perfect repeats >10 nucleotides and ?20 nucleotides) was one SSR per 25.6 kb. The dynamics of SSRs will be a powerful tool for taxonomic, phylogenetic, genome mapping and population genetic studies as SSR based markers show high levels of allelic variation, codominant inheritance and ease of analysis.
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G708E mutation in the androgen receptor results in complete loss of androgen function.
J. Androl.
PUBLISHED: 07-29-2010
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End-organ resistance to androgens, called androgen insensitivity syndrome (AIS), is a rare disorder. The most common cause of AIS is mutations(s) in the androgen receptor (AR) gene; however, a significant number of these mutations have not been functionally analyzed. In the present study, we analyzed a case of complete AIS for mutations in the AR gene. Sequencing of the entire coding region of the AR gene revealed a 2650G>A mutation (mRNA sequence reference) in exon 4 of the gene, resulting in replacement of glycine with glutamate at codon 708 in the ligand-binding domain of the AR protein. The mutation was absent in 200 normal male individuals analyzed to look at its occurrence in general population. In vitro androgen-binding and transactivation assays showed that the mutation resulted in approximately 65% loss of ligand binding and almost complete loss of transactivation function. Complete AIS in this individual was due to a G708E substitution in the AR protein.
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Role of ethnic variations in TNF-? and TNF-? polymorphisms and risk of breast cancer in India.
Breast Cancer Res. Treat.
PUBLISHED: 07-28-2010
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TNF-? and -?, the multi-functional pro-inflammatory cytokines, are known to play important roles in both tumor progression and destruction based on their concentrations. Growth factors and various stimuli such as cytokines regulate proliferation of the breast epithelial cells. Therefore, the polymorphisms in the genes encoding these signaling molecules could affect the risk of breast cancer. We have investigated selected genetic polymorphisms in TNF-? promoter (rs1800629, -308 G>A and rs361525, -238 G>A) and TNF-? intron 1 (rs909253, +252 A>G) in ethnically two different case-control groups from India. The study included 200 cases and 200 controls from an Indo-European (North Indian) group, and 265 cases and 237 controls from a Dravidian (South Indian) group. Genotyping of a total of 902 individuals was done by direct DNA sequencing. None of the polymorphisms showed significant association with breast cancer in the Indo-European group; however, all the three polymorphisms showed strong association with breast cancer in the Dravidian group. Further, sub-group analysis in the Indo-European group showed no significant difference between pre-menopausal cases and controls or between post-menopausal cases and controls at any of the loci analyzed. However, all the polymorphisms in the Dravidian group were significantly associated with pre-menopausal but not with post-menopausal breast cancer. In conclusion, TNF-? and -? polymorphisms are strongly associated with breast cancer in the Dravidian but not in the Indo-European group.
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Nature versus nurture--plant resources in management of male infertility.
Front Biosci (Elite Ed)
PUBLISHED: 06-03-2010
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Male infertility, apart from being a multi-factorial disorder, has no defined etiology in almost half of the infertile men. The complex etiology demands a complex remedy which can heal several ailments together. Currently available specific treatments are largely inefficient in infertility treatment. Medicinal plants present a repertoire capable of providing varied constituents which could be helpful in infertility management. However, the literature on the same is scanty and we have not explored even 1 percent of the available plant resources. Herein, we present a systematic review of clinical and experimental data on the use of Indian medicinal herbs in the treatment of male infertility. Literature suggests that most of the medicinal herbs exhibit a three dimensional effect of reducing oxidative/psychological stress, fatigue and promoting libido. This review is oriented to identify and highlight aphrodisiac, adaptogenic, anti-oxidant and nutritional properties of these plants and aims at promoting exploration of these valuable medicinal resources.
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CAG repeat length polymorphism in the androgen receptor gene and breast cancer risk: data on Indian women and survey from the world.
Breast Cancer Res. Treat.
PUBLISHED: 05-10-2010
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We analyzed the length of the CAG repeats of the androgen receptor gene in Indian women with breast cancer, and compared the data with that of other populations across the world in an attempt to find a potential pattern of association. The study was undertaken on 1,408 individuals comprising 747 breast cancer patients and 661 control individuals recruited from three southern states of India: Andhra Pradesh, Tamil Nadu, and Karnataka. The comparison revealed no difference in mean length of the repeat between cases and controls in any of the three groups or in the analysis of pooled data. No significant difference between pre- and post-menopausal cases in any of the three groups or in the analysis of pooled data was observed. Most of the studies to date support either positive association (longer repeats--increased disease risk) or no association, and only 2 out of 20 studies reported negative association (inverse correlation between repeat length and disease risk). Comparison of these data with those from other populations revealed several interesting facts. Particularly notable is that repeat length shows association with breast cancer risk in a population-specific manner with most of the studies on American and Canadian women showing positive association, whereas those on Australian and Israeli women showing no association. Only one study had been conducted on other populations including Asians/South Asians; this restricted us from finding any patterns of association in these populations.
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DNA damage and cholinesterase activity in occupational workers exposed to pesticides.
Environ. Toxicol. Pharmacol.
PUBLISHED: 04-27-2010
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The present study was designed to evaluate genotoxicity, acetyl cholinesterase (AChE) activity, hepatic and renal toxicity in occupational workers exposed to mixture of pesticides (n=70) with same number of healthy subjects as controls. The mean comet tail DNA % (TD %) and tail moment (TM) were used to measure DNA damage, while AChE activity and other biochemical parameters such as markers of nephrotoxicity (urea and creatinine) and hepatotoxicity (AST, ALT and ALP) were measured as biomarkers for toxicity due to exposure of pesticides. The occupational workers were continuously exposed to mixture of pirimiphos methyl, chlorpyrifos, temephos and malathion on a regular interval as per usage and activity. The comet assay using lymphocytes of exposed workers showed significantly higher TD percentage value (60.43% vs. 31.86%, p<0.001) and TM value (14.48 ?m vs. 6.42 ?m, p<0.001) in occupational workers as compared to controls. AChE activity in erythrocytes was found to be decreased (3.45 KAU/L vs. 9.55 KAU/L in controls, p<0.001) and associated with the duration of exposure to pesticides used by the workers. Enzyme levels for hepatic and renal functions were also found significantly different in occupational workers than healthy controls (p<0.001). These results suggest that the exposure to mixture of pirimiphos methyl, chlorpyrifos, temephos and malathion may induce DNA damage, decrease in AChE activity, hepatotoxicity as well as nephrotoxicity. Periodic biomonitoring of these biomarkers along with imparting education and training to occupational workers for safe application of pesticides is recommended for its potential hazards.
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C601S mutation in the androgen receptor results in partial loss of androgen function.
J. Steroid Biochem. Mol. Biol.
PUBLISHED: 04-21-2010
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The present study was undertaken on a case of partial androgen insensitivity syndrome to look at the etiology of the disorder. The patient exhibited a female phenotype despite 46,XY chromosome complement. Direct DNA sequencing of coding region of the androgen receptor gene in this case revealed a 2329G>C substitution (cDNA sequence reference) in exon 3 of the gene. The substitution resulted in replacement of Cys with Ser at codon 601 of the ligand-binding domain of the protein. Analyses on 200 control samples revealed absence of this substitution(s). In vitro assays were done using COS-1 cells. The mutation resulted in partial (?40%) loss of ligand-binding and significant (?70%) loss of downstream transactivation function. The mutation was absent in the controls. The findings are particularly interesting since another substitution at the same codon (TGC-TTC) has been reported in association with complete androgen insensitivity syndrome.
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R659X mutation in the MLH1 gene in hereditary non-polyposis colorectal cancer(HNPCC) in an Indian extended family.
Indian J. Med. Res.
PUBLISHED: 02-20-2010
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Hereditary non-polyposis colorectal cancer (HNPCC or Lynch syndrome), is a genetically heterogeneous disorder that is believed to account for 2-10 per cent of all the colorectal cancer cases. The disease follows autosomal dominant inheritance pattern with high penetrance (85%) and younger age of onset when compared to patients with sporadic tumours. HNPCC is associated with germ-line mutations in the DNA mismatch repair (MMR) genes namely MLH1, MSH2, MSH6, and PMS2. The present study was aimed at analyzing mismatch repair gene(s) in an extended Indian family satisfying the Amsterdam criteria, and extending the analysis to general population to estimate frequency of the mutations/polymorphisms observed.
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Mitochondria, spermatogenesis and male infertility.
Mitochondrion
PUBLISHED: 02-15-2010
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Mitochondria and its role in male reproduction has remained an enigma since long. Similarly, etiology of male infertility in a large percentage of individuals, mainly primary infertility, has evaded concrete conclusions. Oxidative metabolism, energy production and free radical generation are the principal biological reactions occurring inside mitochondria. In addition to the above, mitochondria participates in an important process of apoptosis. Mitochondrial causes of infertility have triggered interest because of its presence in the tail of sperm and immense need of energy for sperm motility. Several studies on mitochondria have strongly suggested its role in fertility, some of which support mitochondrial role presenting numerous hypotheses, whereas others deny its very existence as a causative factor. We have, in the present review of existing literature, covered the role of mitochondria right from spermatogenesis to male infertility.
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Comprehensive metabolic fingerprinting of Withania somnifera leaf and root extracts.
Phytochemistry
PUBLISHED: 02-12-2010
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Profiling of metabolites is a rapidly expanding area of research for resolving metabolic pathways. Metabolic fingerprinting in medicinally important plants is critical to establishing the quality of herbal medicines. In the present study, metabolic profiling of crude extracts of leaf and root of Withania somnifera (Ashwagandha), an important medicinal plant of Indian system of medicine (ISM) was carried out using NMR and chromatographic (HPLC and GC-MS) techniques. A total of 62 major and minor primary and secondary metabolites from leaves and 48 from roots were unambiguously identified. Twenty-nine of these were common to the two tissues. These included fatty acids, organic acids, amino acids, sugars and sterol based compounds. Eleven bioactive sterol-lactone molecules were also identified. Twenty-seven of the identified metabolites were quantified. Highly significant qualitative and quantitative differences were noticed between the leaf and root tissues, particularly with respect to the secondary metabolites.
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Withania somnifera Improves Semen Quality in Stress-Related Male Fertility.
Evid Based Complement Alternat Med
PUBLISHED: 10-01-2009
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Stress has been reported to be a causative factor for male infertility. Withania somnifera has been documented in Ayurveda and Unani medicine system for its stress-combating properties. However, limited scientific literature is available on this aspect of W. somnifera. We undertook the present study to understand the role of stress in male infertility, and to test the ability of W. somnifera to combat stress and treat male infertility. We selected normozoospermic but infertile individuals (N = 60), further categorized in three groups: normozoospermic heavy smokers (N = 20), normozoospermics under psychological stress (N = 20) and normozoospermics with infertility of unknown etiology (N = 20). Normozoospermic fertile men (N = 60) were recruited as controls. The subjects were given root powder of W. somnifera at a rate of 5 g/day for 3 months. Measuring various biochemical and stress parameters before and after treatment, suggested a definite role of stress in male infertility and the ability of W. somnifera to treat stress-related infertility. Treatment resulted in a decrease in stress, improved the level of anti-oxidants and improved overall semen quality in a significant number of individuals. The treatment resulted in pregnancy in the partners of 14% of the patients.
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Mucuna pruriens improves male fertility by its action on the hypothalamus-pituitary-gonadal axis.
Fertil. Steril.
PUBLISHED: 08-28-2009
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To understand the mechanism of action of Mucuna pruriens in the treatment of male infertility.
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Longer CAG repeat length in the androgen receptor gene is associated with premature ovarian failure.
Hum. Reprod.
PUBLISHED: 08-14-2009
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Premature ovarian failure (POF) is a disorder characterized by lack of ovulation and elevated serum gonadotrophin levels before the age of 40 years. The cause of POF in most cases is unknown. As mice lacking the Androgen receptor (Ar) gene reportedly have a POF-like phenotype, we hypothesize that, variations in the AR gene maybe one of the causative factors for POF in humans. Thus the objective of the study is to evaluate the number of CAG repeats in exon 1 of the AR gene in non-familial, non-syndromic cases of POF.
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Longer (TA)n repeat but not A49T and V89L polymorphisms in SRD5A2 gene may confer prostate cancer risk in South Indian men.
J. Androl.
PUBLISHED: 05-14-2009
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Testosterone is converted to 5 alpha-dihydrotestosterone (DHT) by 5 alpha-reductase enzyme, which is encoded by the SRD5A2 gene. DHT is the main androgen responsible for prostate growth. We have analyzed the complete coding region of the SRD5A2 gene in 87 histologically confirmed prostate cancer (PC) patients, 40 benign prostatic hyperplasia (BPH) cases, and 96 control samples from southern parts of India. The study revealed the A49T site to be monomorphic, the V89L site to be highly polymorphic, and the (TA)(n) repeat site to be polymorphic with only 2 alleles in our populations. The distribution of V89L alleles between PC cases and controls was not significantly different; however, (TA)(9) alleles distributed differently between the 2 groups. BPH cases exhibited alleles similar to controls at all polymorphic sites. The sequencing of the whole coding region did not reveal any other known or novel polymorphism in this gene. Our study emphasizes that the (TA)(9) allele might confer certain PC risk but that A49T and V89L polymorphisms do not confer PC risk in South Indian men.
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Hormonal treatment of male infertility: promises and pitfalls.
J. Androl.
PUBLISHED: 05-01-2009
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Approximately 50% of infertility issues are attributable to male factors. A number of different factors may result in similar reductions of sperm count or motility and affect sperm morphology. Not only is the etiology of male infertility difficult to understand, but it is equally challenging to treat male infertility because of its etiological heterogeneity. Because of complex and incomplete knowledge of the underlying causes, most infertile men are described as idiopathically oligozoospermic and/or asthenozoospermic. Different hormonal treatments have been attempted, aiming to improve mainly endogenous follicle-stimulating hormone and/or androgen levels and subsequent spermatogenesis. Various studies have tried to treat infertility through natural pregnancies or increased sperm retrieval for in vitro fertilization techniques, or by treating spermatozoa in vitro to improve its fertilizing potential. The present review focuses on all of the aspects of male infertility treatment by hormone supplementation.
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Withania somnifera improves semen quality by regulating reproductive hormone levels and oxidative stress in seminal plasma of infertile males.
Fertil. Steril.
PUBLISHED: 04-03-2009
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To investigate the impact of Withania somnifera roots on semen profile, oxidative biomarkers, and reproductive hormone levels of infertile men.
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Ala 586 Asp mutation in androgen receptor disrupts transactivation function without affecting androgen binding.
Fertil. Steril.
PUBLISHED: 03-20-2009
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To understand the pathogenesis of the androgen insensitivity syndrome.
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Courveilhier baumgarten syndrome: a rare syndrome revisited.
Int J Health Sci (Qassim)
PUBLISHED: 01-01-2009
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A 25 year old male had presented with history of abdominal distension of 20 days duration and on examination he had prominent abdominal vein and a venous hum with Ascites. He had no history of altered sensorium, melena. The clinical scenario was suggestive of Cruveilhier Baumgarten syndrome.
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A case of oligoasthenoteratozoospermia with AZFc deletion and persistent oxidative stress.
Indian J Hum Genet
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Y-chromosomal microdeletions are associated with severe oligozoospermia or azoospermia. AZFc microdeletions have been always associated with severe oligozoospermia or azoospermia with a rare occurrence in individuals with other infertility phenotypes. We report here a rare case of an infertile man carrying AZFc deletion, whose semen picture is oligoasthenoteratozoospermia complexed with seminal oxidative stress. Anti-oxidant therapy could make no change in either oxidative stress biomarker levels of semen, seminal parameters or serum hormone levels. Therefore, oligoasthenoteratozoospermia in the present case correlates with AZFc deletion, and high content of abnormal sperm eventually might be responsible for persistently elevated reactive oxygen species levels. Understanding the function of genes in AZFc region could help decipher the exact cause of the phenotype in such cases.
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IL-4 haplotype -590T, -34T and intron-3 VNTR R2 is associated with reduced malaria risk among ancestral indian tribal populations.
PLoS ONE
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Interleukin 4 (IL-4) is an anti-inflammatory cytokine, which regulates balance between T(H)1 and T(H)2 immune response, immunoglobulin class switching and humoral immunity. Polymorphisms in this gene have been reported to affect the risk of infectious and autoimmune diseases.
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Identification of the potential of microbial combinations obtained from spent mushroom cultivation substrates for use in textile effluent decolorization.
Bioresour. Technol.
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The study presents variation in microbial population of Agaricus bisporus, Pleurotus sajor-caju and Volvariella volvacea spent substrates (SMS) along with ligninolytic enzymes activity and textile effluent decolorization potential of microorganisms isolated from these. The effect of temperature, pH, carbon sources and immobilizing agents on effluent decolorization using different combinations of these microorganisms has also been studied. SMS of P. sajor-caju harbored highest population and diversity of bacteria and fungi compared to other SMSs. Schizophyllum commune and Pezizomycotina sp. from P. sajor-caju SMS, exhibited highest activities of laccase (11.8 and 8.32U mL(-1)) and lignin peroxidase (339 and 318 UL(-1)), while Pseudomonas fluorescens of Manganese peroxidase. Highest decolorization was in presence of glucose and sucrose at 30°C, and microbial consortium comprised of the immobilized forms of S. commune and Pezizomycotina sp. on wheat straw and broth cultures of P. fluorescens, Bacillus licheniformis and Bacillus pumilus.
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Cloning and functional characterization of 3-hydroxy-3-methylglutaryl coenzyme A reductase gene from Withania somnifera: an important medicinal plant.
Protoplasma
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Withania somnifera (L.) Dunal is one of the most valuable medicinal plants synthesizing a large number of pharmacologically active secondary metabolites known as withanolides, the C28-steroidal lactones derived from triterpenoids. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the biosynthetic pathway and genes responsible for biosynthesis of these compounds. In this study, we have characterized the gene encoding 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR; EC 1.1.1.34) catalyzing the key regulatory step of the isoprenoid biosynthesis. The 1,728-bp full-length cDNA of Withania HMGR (WsHMGR) encodes a polypeptide of 575 amino acids. The amino acid sequence homology and phylogenetic analysis suggest that WsHMGR has typical structural features of other known plant HMGRs. The relative expression analysis suggests that WsHMGR expression varies in different tissues as well as chemotypes and is significantly elevated in response to exposure to salicylic acid, methyl jasmonate, and mechanical injury. The functional color assay in Escherichia coli showed that WsHMGR could accelerate the biosynthesis of carotenoids, establishing that WsHMGR encoded a functional protein and may play a catalytic role by its positive influence in isoprenoid biosynthesis.
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Polymorphic variations in IL-1?, IL-6 and IL-10 genes, their circulating serum levels and breast cancer risk in Indian women.
Cytokine
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Cytokines are known as important regulators of the entire gamut of cancer from initiation, invasion and metastasis. This fact and plethora of gene polymorphism data prompted us to investigate cytokine gene polymorphisms in breast cancer (BC) patients.
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Null genotypes at the GSTM1 and GSTT1 genes and the risk of benign prostatic hyperplasia: a case-control study and a meta-analysis.
Prostate
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Benign prostatic hyperplasia (BPH) is an age related non-malignant disease diagnosed as lower urinary tract symptoms and prostatic enlargement. Null genotypes in drug detoxification glutathione-S-transferase genes/enzymes, such as GSTT1 and GSTM1 have been reported to increase risk of several cancers including prostate. Meta-analysis on PC also suggested significant impact of GSTM1 null genotype but not that of GSTT1; however, BPH data have not been subjected to meta-analysis.
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Small RNA in spermatogenesis and male infertility.
Front Biosci (Schol Ed)
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Small RNA has become a crucial regulator of protein synthesis during spermatogenesis. Alterations in small RNA function prove to be detrimental to proper spermatogenesis. As many patients suffer from idiopathic infertility, understanding the molecular mechanisms of small RNA identifies possible causes of certain types of infertility. With a comprehensive review of the history of miRNA and piRNA function and specificity in the testis from a wide range of studies offers a view of detrimental defects of small RNA. By combining a concise overview of small RNA mechanism and recent research we explain how some cases of male infertility can be a product of complications in specific small RNA functions. The future direction section offers insight into how infertility treatment may be approached with a novel perspective.
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Cloning and characterization of 2-C-methyl-D-erythritol-4-phosphate pathway genes for isoprenoid biosynthesis from Indian ginseng, Withania somnifera.
Protoplasma
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Withania somnifera (L.) is one of the most valuable medicinal plants used in Ayurvedic and other indigenous medicines. Pharmaceutical activities of this herb are associated with presence of secondary metabolites known as withanolides, a class of phytosteroids synthesized via mevalonate (MVA) and 2-C-methyl-D-erythritol-4-phosphate pathways. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized two genes encoding 1-deoxy-D-xylulose-5-phosphate synthase (DXS; EC 2.2.1.7) and 1-deoxy-D-xylulose-5-phosphate reductase (DXR; EC 1.1.1.267) enzymes involved in the biosynthesis of isoprenoids. The full-length cDNAs of W. somnifera DXS (WsDXS) and DXR (WsDXR) of 2,154 and 1,428 bps encode polypeptides of 717 and 475 amino acids residues, respectively. The expression analysis suggests that WsDXS and WsDXR are differentially expressed in different tissues (with maximal expression in flower and young leaf), chemotypes of Withania, and in response to salicylic acid, methyl jasmonate, as well as in mechanical injury. Analysis of genomic organization of WsDXS shows close similarity with tomato DXS in terms of exon-intron arrangements. This is the first report on characterization of isoprenoid biosynthesis pathway genes from Withania.
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Ion channels in sperm physiology and male fertility and infertility.
J. Androl.
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Ion channels regulate the membrane potential and intracellular ionic concentration and thus serve a central role in various cellular processes. Several ion channels have been identified in the germ cells, including sperm, emphasizing their importance in male fertility and reproduction. The molecular mechanism of ion transport and the nature of the ion channels involved have begun to emerge only recently despite the fact that several ligand-gated and voltage-gated channels have been identified and localized on sperm. The presence of the sperm-associated cation channel (CatSper1-4) gene family, proton voltage-gated ion channel (Hv1), potassium voltage-gated ion channel (SLO3/KCNU1), sodium voltage-gated channel (NaV1.1-1.9), and the members of the transient receptor potential (TRP) channel family suggest an indispensable role for ion channels in sperm physiology and fertility potential. Ion channels are the key players in very important processes such as capacitation and the acrosome reaction, which are critical steps in sperm physiology preparing for fertilization. For example, CatSper, Hv1, SLO3, and TRP channel family members have been proposed to participate in the acrosome reaction, thereby making them most important for sperm fertility. Similarly, NaV channels could play a crucial role in noncapacitated sperm and in the initial capacitation steps. The role of ion channels seems indispensable for sperm fertility as evidenced by studies on animal models; however, the functional defects in infertile human males await further exploration. This article represents an update on the role of ion channels in sperm physiology, male fertility, and infertility.
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Physical deformities relevant to male infertility.
Nat Rev Urol
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Infertile men are frequently affected by physical abnormalities that might be detected on routine general and genital examinations. These structural abnormalities might damage or block the testes, epididymis, seminal ducts or other reproductive structures and can ultimately decrease fertility. Physical deformities are variable in their pathological impact on male reproductive function; some render men totally sterile, such as bilateral absence of the vasa deferentia, while others cause only mild alterations in semen parameters. Concise and up-to-date information regarding the contemporary epidemiological characteristics, clinical features and pathophysiological impacts of these common abnormalities on male fertility is crucial for the practicing urologist to identify the best treatment option.
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