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Find video protocols related to scientific articles indexed in Pubmed.
A case of adult dermatomyositis with calcinosis universalis.
Indian J Nucl Med
PUBLISHED: 11-18-2014
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Calcinosis, although frequent in juvenile dermatomyositis is a rare finding in adults. It is more common in later phases of the disease, involving sites under chronic stress and trauma. We present a 52-year-old female patient of dermatomyositis who on single-photon emission computed tomography/computed tomography hybrid images showed exclusive subcutaneous fat calcinosis, also known as calcinosis universalis - a phenomenon that is only rarely reported in adult-onset dermatomyositis.
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Enhancement of positron emission tomography-computed tomography image quality using the principle of stochastic resonance.
Indian J Nucl Med
PUBLISHED: 11-18-2014
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Acquisition of higher counts improves visual perception of positron emission tomography-computed tomography (PET-CT) image. Larger radiopharmaceutical doses (implies more radiation dose) are administered to acquire this count in a short time period. However, diagnostic information does not increase after a certain threshold of counts. This study was conducted to develop a post processing method based on principle of "stochastic resonance" to improve visual perception of the PET-CT image having a required threshold counts.
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Development of dog mammary tumor xenograft in immunosuppressed Swiss albino mice.
Indian J. Exp. Biol.
PUBLISHED: 10-28-2014
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Development and study of dog mammary tumour xenograft in immunosuppressed Swiss Albino Mice adds a new dimension in cancer research as dog tumors have many similarities with human tumors regarding progression, histopathology, molecular mechanism, immune response and therapy. Failure of the immune system to recognize and eliminate cancer cells leads to cancer progression and the fight between immune cells and cancer cells has a great role in understanding the mechanism of cancer progression and elimination. Rejection and acceptance of tumour xenograft depends on efficiency of CD4+, CD8+ and NK cell populations. In the present investigation, dog mammary tumor xenograft in cyclosporine-A and gamma-irradiated, immunosuppressed Swiss Albino mice was developed and the immune cell status of graft accepted and rejected mice was assessed. It was observed that all the major immune cells (CD4+, CD8+ and NK cells) play an equal role in tumour rejection.
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Electrophysiological evidence for enhanced representation of food stimuli in working memory.
Exp Brain Res
PUBLISHED: 10-17-2014
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Studies from our laboratory have shown that, relative to neutral objects, food-related objects kept in working memory (WM) are particularly effective in guiding attention to food stimuli (Higgs et al. in Appetite, 2012). Here, we used electrophysiological measurements to investigate the neural representation of food versus non-food items in WM. Subjects were presented with a cue (food or non-food item) to either attend to or hold in WM. Subsequently, they had to search for a target, while the target and distractor were each flanked by a picture of a food or non-food item. Behavioural data showed that a food cue held in WM modulated the deployment of visual attention to a search target more than a non-food cue, even though the cue was irrelevant for target selection. Electrophysiological measures of attention, memory and retention of memory (the P3, LPP and SPCN components) were larger when food was kept in WM, compared to non-food items. No such effect was observed in a priming task, when the initial cue was merely identified. Overall, our electrophysiological data are consistent with the suggestion that food stimuli are particularly strongly represented in the WM system.
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Activation of p53-dependent/-independent pathways of apoptotic cell death by chelerythrine in a murine T cell lymphoma.
Leuk. Lymphoma
PUBLISHED: 10-14-2014
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Abstract The p53 tumor suppressor protein has been implicated as an activator of apoptosis. In order to investigate the effect of chelerythrine and staurosporine on the activation of p53-dependent/-independent pathways of DL cells death, cells were treated with chelerythrine and staurosporine for 1hr, 3hr and 6hr respectively. It was found that treatment with chelerythrine and staurosporine increased the expression of total-p53/phospho-53 (ser-15) significantly at protein and mRNA levels which results in activation of p53-dependent apoptotic pathway in DL cells. In addition, increase activities of cyt-c, caspase-9 and caspase-3 and degradation of DNA into fragments were confirmed activation of p53-independent apoptotic pathway in p53 knockdown RNAi-DL cells. In brief, the present report focused on the activation of p53-dependent/-independent apoptotic pathways in DL cells. Therefore, targeting of p53-dependent/-independent apoptotic pathway may arose the possibility of designing and developing better therapeutic regimen to treat Dalton's lymphoma and other human cancers.
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Vitamin D status and risk of ischemic stroke in North Indian patients.
Indian J Endocrinol Metab
PUBLISHED: 10-07-2014
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Accumulating evidence suggests that vitamin D deficiency is associated with increased risk of stroke. Contributing mechanisms have been linked to the association of vitamin D deficiency with the presence of hypertension, diabetes mellitus, and atherosclerosis, however, the evidence is conflicting.
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Single-cell Western blotting after whole-cell imaging to assess cancer chemotherapeutic response.
Anal. Chem.
PUBLISHED: 10-01-2014
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Intratumor heterogeneity remains a major obstacle to effective cancer therapy and personalized medicine. Current understanding points to differential therapeutic response among subpopulations of tumor cells as a key challenge to successful treatment. To advance our understanding of how this heterogeneity is reflected in cell-to-cell variations in chemosensitivity and expression of drug-resistance proteins, we optimize and apply a new targeted proteomics modality, single-cell western blotting (scWestern), to a human glioblastoma cell line. To acquire both phenotypic and proteomic data on the same, single glioblastoma cells, we integrate high-content imaging prior to the scWestern assays. The scWestern technique supports thousands of concurrent single-cell western blots, with each assay comprised of chemical lysis of single cells seated in microwells, protein electrophoresis from those microwells into a supporting polyacrylamide (PA) gel layer, and in-gel antibody probing. We systematically optimize chemical lysis and subsequent polyacrylamide gel electrophoresis (PAGE) of the single-cell lysate. The scWestern slides are stored for months then reprobed, thus allowing archiving and later analysis as relevant to sparingly limited, longitudinal cell specimens. Imaging and scWestern analysis of single glioblastoma cells dosed with the chemotherapeutic daunomycin showed both apoptotic (cleaved caspase 8- and annexin V-positive) and living cells. Intriguingly, living glioblastoma subpopulations show up-regulation of a multidrug resistant protein, P-glycoprotein (P-gp), suggesting an active drug efflux pump as a potential mechanism of drug resistance. Accordingly, linking of phenotype with targeted protein analysis with single-cell resolution may advance our understanding of drug response in inherently heterogeneous cell populations, such as those anticipated in tumors.
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Effect of electromagnetic irradiation produced by 3G mobile phone on male rat reproductive system in a simulated scenario.
Indian J. Exp. Biol.
PUBLISHED: 09-23-2014
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Reports of declining male fertility have renewed interest in assessing the role of electromagnetic fields (EMFs). Testicular function is particularly susceptible to the radiation emitted by EMFs. Significant decrease in sperm count, increase in the lipid peroxidation damage in sperm cells, reduction in seminiferous tubules and testicular weight and DNA damage were observed following exposure to EMF in male albino rats. The results suggest that mobile phone exposure adversely affects male fertility.
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Platelet mechanosensing of substrate stiffness during clot formation mediates adhesion, spreading, and activation.
Proc. Natl. Acad. Sci. U.S.A.
PUBLISHED: 09-22-2014
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As platelets aggregate and activate at the site of vascular injury to stem bleeding, they are subjected to a myriad of biochemical and biophysical signals and cues. As clot formation ensues, platelets interact with polymerizing fibrin scaffolds, exposing platelets to a large range of mechanical microenvironments. Here, we show for the first time (to our knowledge) that platelets, which are anucleate cellular fragments, sense microenvironmental mechanical properties, such as substrate stiffness, and transduce those cues into differential biological signals. Specifically, as platelets mechanosense the stiffness of the underlying fibrin/fibrinogen substrate, increasing substrate stiffness leads to increased platelet adhesion and spreading. Importantly, adhesion on stiffer substrates also leads to higher levels of platelet activation, as measured by integrin ?IIb?3 activation, ?-granule secretion, and procoagulant activity. Mechanistically, we determined that Rac1 and actomyosin activity mediate substrate stiffness-dependent platelet adhesion, spreading, and activation to different degrees. This capability of platelets to mechanosense microenvironmental cues in a growing thrombus or hemostatic plug and then mechanotransduce those cues into differential levels of platelet adhesion, spreading, and activation provides biophysical insight into the underlying mechanisms of platelet aggregation and platelet activation heterogeneity during thrombus formation.
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Transcriptome sequencing of rhizome tissue of Sinopodophyllum hexandrum at two temperatures.
BMC Genomics
PUBLISHED: 09-17-2014
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Sinopodophyllum hexandrum is an endangered medicinal herb, which is commonly present in elevations ranging between 2,400-4,500 m and is sensitive to temperature. Medicinal property of the species is attributed to the presence of podophyllotoxin in the rhizome tissue. The present work analyzed transcriptome of rhizome tissue of S. hexandrum exposed to 15°C and 25°C to understand the temperature mediated molecular responses including those associated with podophyllotoxin biosynthesis.
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Predictors of atypical femoral fractures during long term bisphosphonate therapy: a case series & review of literature.
Indian J. Med. Res.
PUBLISHED: 09-16-2014
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Bisphosphonates (BPs) are the most widely prescribed medicines for the treatment of osteoporosis because of their efficacy and favourable safety profile. There have been, several reports on an increased incidence of atypical femoral fractures after long term treatment with BPs. The objective of this study was to evaluate the clinical presentation including prodromal symptoms, skeletal radiograph findings, type and duration of BPs received and treatment outcome of patients who developed atypical femoral fractures during bisphosphonate therapy.
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The nature and pattern of coronary stent recalls.
J Invasive Cardiol
PUBLISHED: 09-09-2014
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Each year, over 1 million percutaneous coronary interventions (PCIs) are performed in the United States. Coronary stents have been shown to reduce restenosis or abrupt vessel closure and therefore have improved the success of PCI. Rarely, manufacturers recall stents due to unanticipated problems. We sought to study the extent and pattern of stent recall.
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Bone defect repair in mice by mesenchymal stem cells.
Methods Mol. Biol.
PUBLISHED: 09-01-2014
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Adult bone marrow niche contains rare primitive but highly functional multipotent progenitors (e.g., mesenchymal stem cells; MSCs) capable of differentiating into specific mesenchymal tissues like bone, cartilage, muscle, fat tissues, ligament, dermis, bone marrow stroma, tendon, and other connective tissues. Upon in vivo transplantation, MSCs also secrete a wide range of growth factors, immunomodulatory cytokines, and important bioactive macromolecules including cell-derived exosomes to structure regenerative microenvironments. This protocol describes a mouse model to study bone formation/regeneration from adult mesenchymal stem cells.
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Engineering strategies to mimic the glioblastoma microenvironment.
Adv. Drug Deliv. Rev.
PUBLISHED: 08-29-2014
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Glioblastoma multiforme (GBM) is the most common and deadly brain tumor, with a mean survival time of only 21months. Despite the dramatic improvements in our understanding of GBM fueled by recent revolutions in molecular and systems biology, treatment advances for GBM have progressed inadequately slowly, which is due in part to the wide cellular and molecular heterogeneity both across tumors and within a single tumor. Thus, there is increasing clinical interest in targeting cell-extrinsic factors as way of slowing or halting the progression of GBM. These cell-extrinsic factors, collectively termed the microenvironment, include the extracellular matrix, blood vessels, stromal cells that surround tumor cells, and all associated soluble and scaffold-bound signals. In this review, we will first describe the regulation of GBM tumors by these microenvironmental factors. Next, we will discuss the various in vitro approaches that have been exploited to recapitulate and model the GBM tumor microenvironment in vitro. We conclude by identifying future challenges and opportunities in this field, including the development of microenvironmental platforms amenable to high-throughput discovery and screening. We anticipate that these ongoing efforts will prove to be valuable both as enabling tools for accelerating our understanding of microenvironmental regulation in GBM and as foundations for next-generation molecular screening platforms that may serve as a conceptual bridge between traditional reductionist systems and animal or clinical studies.
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Bilateral congenital absence of flexor pollicis longus with thumb hypoplasia and thenar atrophy.
Indian J Radiol Imaging
PUBLISHED: 08-13-2014
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Congenital absence of flexor pollicis longus with or without associated anomalies of thenar muscles and thumb is of rare occurrence. Inability to flex the interphalangeal joint of the thumb and absent dorsal wrinkles and flexion creases of the thumb are important clues to the diagnosis. Routine radiography and cross-sectional imaging help to confirm and document the condition. This article presents an extremely rare case of bilateral congenital absence of flexor pollicis longus tendon with thumb hypoplasia and thenar atrophy.
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Matrix regulation of tumor-initiating cells.
Prog Mol Biol Transl Sci
PUBLISHED: 08-02-2014
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The recognition that the progression of many tumors may be driven by specific subpopulations of cells with stem/progenitor-like properties (tumor-initiating cells or TICs, a.k.a. cancer stem cells) represents an important recent paradigm shift in cancer biology and therapeutics. TICs in solid tissues are expected to interface with the extracellular matrix (ECM), which can strongly influence cell behavior through a variety of biochemical and biophysical mechanisms. Understanding ECM regulation of TIC behavior is important for developing strategies to isolate, expand, and characterize TICs in a laboratory setting and for understanding the roles ECM-based inputs may play in disease progression and therapy. In this chapter, we discuss how the ECM regulates TICs, starting with a brief overview of TIC biology, isolation, and characterization, molecular mechanisms through which TICs may be regulated by ECM-based signals, and the potential importance of these signals to TIC-driven tumor progression and metastasis.
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Src-mediated post-translational regulation of endoglin stability and function is critical for angiogenesis.
J. Biol. Chem.
PUBLISHED: 07-28-2014
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Endoglin is a transforming growth factor ? (TGF-?) co-receptor essential for angiogenesis and tumor vascularization. Endoglin modulates the crucial balance between pro- and anti-angiogenic signaling by activin receptor-like kinase (ALK) 1, 5, and TGF-? type II (T?RII) receptors. Despite its established role in physiology and disease, the mechanism of endoglin down-regulation remains unknown. Here we report that the conserved juxtamembrane cytoplasmic tyrosine motif ((612)YIY(614)) is a critical determinant of angiogenesis. Src directly phosphorylates this motif to induce endoglin internalization and degradation via the lysosome. We identified epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) as Src-activators that induce endoglin turnover following (612)YIY(614) phosphorylation. Interestingly, Src phosphorylation of endoglin-(612)YIY(614) was also an important process for receptor down-regulation by TRACON105 (TRC105), an endoglin-targeting antibody currently in clinical trials. The regulation of (612)YIY(614) phosphorylation was critical for angiogenesis, as both the phosphomimetic and unphosphorylatable mutants impaired endothelial functions including proliferation, migration, and capillary tube formation. Collectively, these findings establish Src and pro-angiogenic mitogens as critical mediators of endoglin stability and function.
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Simultaneous and independent tuning of RhoA and Rac1 activity with orthogonally inducible promoters.
Integr Biol (Camb)
PUBLISHED: 07-22-2014
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The GTPases RhoA and Rac1 are key regulators of cell spreading, adhesion, and migration, and they exert distinct effects on the actin cytoskeleton. While RhoA classically stimulates stress fiber assembly and contraction, Rac1 promotes branched actin polymerization and membrane protrusion. These competing influences are reinforced by antagonistic crosstalk between RhoA and Rac1, which has complicated efforts to identify the specific mechanisms by which each GTPase regulates cell behavior. We therefore wondered whether RhoA and Rac1 are intrinsically coupled or whether they can be manipulated independently. To address this question, we placed constitutively active (CA) RhoA under a doxycycline-inducible promoter and CA Rac1 under an orthogonal cumate-inducible promoter, and we stably introduced both constructs into glioblastoma cells. We found that doxycycline addition increased RhoA activity without altering Rac1, and similarly cumate addition increased Rac1 activity without altering RhoA. Furthermore, co-expression of both mutants enabled high activation of RhoA and Rac1 simultaneously. When cells were cultured on collagen hydrogels, RhoA activation prevented cell spreading and motility, whereas Rac1 activation stimulated migration and dynamic cell protrusions. Interestingly, high activation of both GTPases induced a third phenotype, in which cells migrated at intermediate speeds similar to control cells but also aggregated into large, contractile clusters. In addition, we demonstrate dynamic and reversible switching between high RhoA and high Rac1 phenotypes. Overall, this approach represents a unique way to access different combinations of RhoA and Rac1 activity levels in a single cell and may serve as a valuable tool for multiplexed dissection and control of mechanobiological signals.
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Molecular cloning and expression of high GC-rich novel tumor suppressor gene HIC-1.
Mol. Biotechnol.
PUBLISHED: 07-09-2014
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Hypermethylated in Cancer-1 (HIC-1) is a novel tumor suppressor plays crucial role in tumor formation through loss of function by hypermethylation. HIC-1 is known as transcriptional factor whereas little known about its structure and function. Requirement felt to clone and express full coding protein and reveal various domains and binding pattern onto promoters conducting biophysical studies which lack in current scenario. Production of sufficient amounts of protein is frequent bottleneck in structural biology. Cloning full-length HIC-1 with >73 % GC content poses a daunting task with sequencing and expression adds more to the challenge. We describe the methodology for specific amplification, cloning, sequencing, and expression of HIC-1 in E. coli. Standardization using 1.5 U pfu polymerase in (NH4)2SO4 containing buffer gave specific amplification with 10 % DMSO and 1.5 mM MgCl2. Sequencing achieved using base analog 7-de aza dGTP (0.2 mM) or denaturant like DMSO (10 %) or betaine (1 M). Expression using strains of E. coli induced by different concentrations of IPTG (0.5-5.0 mM) for time points of 4, 8, 16, 20, and 24 h at different temperatures 25, 30, and 37 °C. Full-length clone successfully expressed in BL21-Codon Plus-RP using 1 mM concentration of IPTG for 8 h at 37 °C gave prominent band of 74 kDa.
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Effect of Vitamin D Replacement on Hemoglobin Concentration in Subjects with Concurrent Iron-Deficiency Anemia and Vitamin D Deficiency: A Randomized, Single-Blinded, Placebo-Controlled Trial.
Acta Haematol.
PUBLISHED: 07-08-2014
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Background: The effect of vitamin D replacement on hemoglobin (Hb) concentration in subjects with concurrent deficiencies of vitamin D and iron is not known. Methods: We report on an investigator-initiated, randomized, single-blinded, placebo-controlled, 12-week interventional trial. Thirty subjects with iron-deficiency anemia (serum ferritin <15 µg/l) were randomized to an intervention arm (cholecalciferol, i.e. vitamin D3, 0.6 million units i.m. once) or placebo. In all subjects, iron deficiency was corrected with parental iron. Other causes of anemia were excluded with appropriate investigation. The primary end point was a rise in Hb concentration. Results: Baseline parameters of age, BMI, hemogram values and levels of serum ferritin, 25-hydroxyvitamin D [25(OH)D] and parathyroid hormone (PTH) were similar in the 2 arms. Twelve weeks after vitamin D replacement, there was a significant increase in 25 (OH)D levels (57.7 ± 20.5 vs. 14.1 ± 6.2 ng/ml, p < 0.0001) and a decrease in PTH levels (32.4 ± 16.4 vs. 52.9 ± 18.4 pg/ml, p = 0.003) in subjects in the intervention arm when compared to the placebo arm. However, the increments in serum ferritin and Hb concentration in the intervention and placebo arm did not differ. Conclusion: Vitamin D replacement in subjects with iron-deficiency anemia after iron correction does not improve Hb concentration further. © 2014 S. Karger AG, Basel.
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Polychlorinated Biphenyls in Residential Soils and their Health Risk and Hazard in an Industrial City in India.
J Public Health Res
PUBLISHED: 07-02-2014
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Polychlorinated biphenyls (PCBs) have never been produced in India, but were used in industrial applications. PCBs have been detected in environmental samples since 1966, and their sources in soils come from depositions of industrial applications, incinerators and biomass combustions. PCBs adsorb to soil particles and persist for long time due to their properties. Their close proximity may also lead to human exposure through ingestion, inhalation, dermal contact, and may exert neurotoxic, mutagenic and carcinogenic health effects.
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A composite hydrogel platform for the dissection of tumor cell migration at tissue interfaces.
Biomaterials
PUBLISHED: 06-24-2014
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Glioblastoma multiforme (GBM), the most prevalent primary brain cancer, is characterized by diffuse infiltration of tumor cells into brain tissue, which severely complicates surgical resection and contributes to tumor recurrence. The most rapid mode of tissue infiltration occurs along blood vessels or white matter tracts, which represent topological interfaces thought to serve as "tracks" that speed cell migration. Despite this observation, the field lacks experimental paradigms that capture key features of these tissue interfaces and allow reductionist dissection of mechanisms of this interfacial motility. To address this need, we developed a culture system in which tumor cells are sandwiched between a fibronectin-coated ventral surface representing vascular basement membrane and a dorsal hyaluronic acid (HA) surface representing brain parenchyma. We find that inclusion of the dorsal HA surface induces formation of adhesive complexes and significantly slows cell migration relative to a free fibronectin-coated surface. This retardation is amplified by inclusion of integrin binding peptides in the dorsal layer and expression of CD44, suggesting that the dorsal surface slows migration through biochemically specific mechanisms rather than simple steric hindrance. Moreover, both the reduction in migration speed and assembly of dorsal adhesions depend on myosin activation and the stiffness of the ventral layer, implying that mechanochemical feedback directed by the ventral layer can influence adhesive signaling at the dorsal surface.
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CD44-Mediated Adhesion to Hyaluronic Acid Contributes to Mechanosensing and Invasive Motility.
Mol. Cancer Res.
PUBLISHED: 06-24-2014
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The high-molecular-weight glycosaminoglycan, hyaluronic acid (HA), makes up a significant portion of the brain extracellular matrix. Glioblastoma multiforme (GBM), a highly invasive brain tumor, is associated with aberrant HA secretion, tissue stiffening, and overexpression of the HA receptor CD44. Here, transcriptomic analysis, engineered materials, and measurements of adhesion, migration, and invasion were used to investigate how HA/CD44 ligation contributes to the mechanosensing and invasive motility of GBM tumor cells, both intrinsically and in the context of Arg-Gly-Asp (RGD) peptide/integrin adhesion. Analysis of transcriptomic data from The Cancer Genome Atlas reveals upregulation of transcripts associated with HA/CD44 adhesion. CD44 suppression in culture reduces cell adhesion to HA on short time scales (0.5-hour postincubation) even if RGD is present, whereas maximal adhesion on longer time scales (3 hours) requires both CD44 and integrins. Moreover, time-lapse imaging demonstrates that cell adhesive structures formed during migration on bare HA matrices are more short lived than cellular protrusions formed on surfaces containing RGD. Interestingly, adhesion and migration speed were dependent on HA hydrogel stiffness, implying that CD44-based signaling is intrinsically mechanosensitive. Finally, CD44 expression paired with an HA-rich microenvironment maximized three-dimensional invasion, whereas CD44 suppression or abundant integrin-based adhesion limited it. These findings demonstrate that CD44 transduces HA-based stiffness cues, temporally precedes integrin-based adhesion maturation, and facilitates invasion.
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Brunner gland hamartoma masquerading as malignancy; a rare case report.
Middle East J Dig Dis
PUBLISHED: 06-16-2014
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Brunner's gland hamartoma is a rare benign tumour of the duodenum. It was first described by Cruveilhier in 1835. Presently around 200 cases have been reported in literature. No sex predilection is seen. Patients usually present in the fifth to sixth decades of life. They may be clinically silent or may present with variable symptoms and occasionally obstructive symptoms and chronic pancreatitis. Endoscopic presentation can be nodular, polypoid or diffuse glandular proliferation with thickening of duodenal wall and hence can be misdiagnosed as malignancy. We describe a case of duodenal tumor reported outside (on biopsy) as well differentiated adenocarcinoma which out as Brunner gland hamartoma upon complete resection. Brunner gland hamartoma may sometimes have a very unusual presentation. Extensive pre-operative evaluation is necessity to avoid radical surgical procedure.
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Natural progression of low-gradient severe aortic stenosis with preserved ejection fraction.
Tex Heart Inst J
PUBLISHED: 06-01-2014
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Because the natural progression of low-gradient aortic stenosis (LGAS) has not been well defined, we performed a retrospective study of 116 consecutive patients with aortic stenosis who had undergone follow-up echocardiography at a median interval of 698 days (range, 371-1,020 d). All patients had preserved left ventricular ejection fraction (>0.50) during and after follow-up. At baseline, patients were classified by aortic valve area (AVA) as having mild stenosis (?1.5 cm(2)), moderate stenosis (?1 to <1.5 cm(2)), or severe stenosis (<1 cm(2)). Severe aortic stenosis was further classified by mean gradient (LGAS, mean <40 mmHg; high-gradient aortic stenosis [HGAS], mean ?40 mmHg). We compared baseline and follow-up values among 4 groups: patients with mild stenosis, moderate stenosis, LGAS, and HGAS. At baseline, 30 patients had mild stenosis, 54 had moderate stenosis, 24 had LGAS, and 8 had HGAS. Compared with the moderate group, the LGAS group had lower AVA but similar mean gradient. Yet the actuarial curves for progressing to HGAS were significantly different: 25% of patients in LGAS reached HGAS status significantly earlier than did 25% of patients in the moderate-AS group (713 vs 881 d; P=0.035). Because LGAS has a high propensity to progress to HGAS, we propose that low-gradient aortic stenosis patients be closely monitored as a distinct subgroup that warrants more frequent echocardiographic follow-up.
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Quercetin and gallic acid mediated synthesis of bimetallic (silver and selenium) nanoparticles and their antitumor and antimicrobial potential.
J Colloid Interface Sci
PUBLISHED: 04-28-2014
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In this study a synthetic approach for the stable, mono-dispersed high yielding bimetallic (Ag-Se) nanoparticles by quercetin and gallic acid is described. The bimetallic nanoparticles were synthesized at room temperature. Different reaction parameters (concentration of quercetin, gallic acid and Ag/Se salt, pH, temperature and reaction time) were optimized to control the properties of nanoparticles. The nanoparticles were characterized by various analytical techniques and their size was determined to be 30-35 nm. Our findings suggest that both the reduction as well as stabilization of nanoparticles were achieved by the flavonoids and phenolics. This study describes the efficacy of quercetin and gallic acid mediated synthesis of bimetallic (Ag-Se) nanoparticles and their in vitro antioxidant, antimicrobial (Gram-positive and Gram-negative bacteria) and antitumor potentials. The synthesized Ag-Se nanoparticles were used as anticancer agents for Dalton lymphoma (DL) cells and in in vitro 80% of its viability was reduced at 50 ?g/mL.
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Stimuli-sensitive intrinsically disordered protein brushes.
Nat Commun
PUBLISHED: 04-19-2014
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Grafting polymers onto surfaces at high density to yield polymer brush coatings is a widely employed strategy to reduce biofouling and interfacial friction. These brushes almost universally feature synthetic polymers, which are often heterogeneous and do not readily allow incorporation of chemical functionalities at precise sites along the constituent chains. To complement these synthetic systems, we introduce a biomimetic, recombinant intrinsically disordered protein that can assemble into an environment-sensitive brush. This macromolecule adopts an extended conformation and can be grafted to solid supports to form oriented protein brushes that swell and collapse dramatically with changes in solution pH and ionic strength. We illustrate the value of sequence specificity by using proteases with mutually orthogonal recognition sites to modulate brush height in situ to predictable values. This study demonstrates that stimuli-responsive brushes can be fabricated from proteins and introduces them as a new class of smart biomaterial building blocks.
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Age-related dysfunction in mechanotransduction impairs differentiation of human mammary epithelial progenitors.
Cell Rep
PUBLISHED: 04-15-2014
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Dysfunctional progenitor and luminal cells with acquired basal cell properties accumulate during human mammary epithelial aging for reasons not understood. Multipotent progenitors from women aged <30 years were exposed to a physiologically relevant range of matrix elastic modulus (stiffness). Increased stiffness causes a differentiation bias towards myoepithelial cells while reducing production of luminal cells and progenitor maintenance. Lineage representation in progenitors from women >55 years is unaffected by physiological stiffness changes. Efficient activation of Hippo pathway transducers YAP and TAZ is required for the modulus-dependent myoepithelial/basal bias in younger progenitors. In older progenitors, YAP and TAZ are activated only when stressed with extraphysiologically stiff matrices, which bias differentiation towards luminal-like phenotypes. In vivo YAP is primarily active in myoepithelia of younger breasts, but localization and activity increases in luminal cells with age. Thus, aging phenotypes of mammary epithelia may arise partly because alterations in Hippo pathway activation impair microenvironment-directed differentiation and lineage specificity.
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Study of incidence and prevalence of musculoskeletal anomalies in a tertiary care hospital of eastern India.
J Clin Diagn Res
PUBLISHED: 04-11-2014
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Congenital anomalies or malformations are anatomical. Structural or functional defects present at birth leads to physical and mental disabilities. With the advent of newer drugs, infectious diseases have taken the backseat and congenital defects have emerged as an important cause of neonatal morbidity and mortality. In India, anomalies of musculoskeletal system have been most commonly reported.
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Effect of gender, biochemical parameters & parathyroid surgery on gastrointestinal manifestations of symptomatic primary hyperparathyroidism.
Indian J. Med. Res.
PUBLISHED: 04-11-2014
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Information on gastrointestinal manifestations and then response after curative parathyroid surgery is scarce in symptomatic primary hyperparathyroidism (PHPT). This study was carried out to analyse gastrointestinal manifestations in patients with PHPT and their associations with biochemical parameters.
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P53 induction accompanying G2/M arrest upon knockdown of tumor suppressor HIC1 in U87MG glioma cells.
Mol. Cell. Biochem.
PUBLISHED: 03-26-2014
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Hypermethylated in cancer 1 (HIC1) is a novel tumor suppressor gene (tsg) frequently silenced by epigenetic modification, predominantly by methylation in different tumors. HIC1 functionally co-operates with p53 in cultured cells as well as in transgenic animals to suppress tumors and has binding site on its promoter. Its over expression often leads to cell cycle arrests. Although HIC1 proven to have role as tsg, its regulation to cell cycle and dependency upon p53 is grossly unknown. In this study, we investigated the role of HIC1 in cell cycle and proliferation of glioma cell line U87MG which has wild type p53, in both serum-containing and serum-deprived medium. Microscopic analysis and MTT assay showed reduced cell number and rate of proliferation upon HIC1 knock down compared to control siRNA (p = 0.025) and untreated cells (p = 0.03) in serum-containing medium and serum-free medium (p = 0.014 vs control siRNA; p = 0.018 vs untreated cells). Cell cycle analysis revealed an arrest at G2/M phase of cell cycle with no demonstrable increase in apoptosis with both medium. An increased expression of p53 concomitant with HIC1 knockdown was observed. Furthermore P21, a p53 responsive gene, along with p27 was significantly increased in comparison with controls. Our results demonstrated an important role of HIC1 for the normal progression of cell cycle, and at molecular level, it could affect the homeostasis of p53 as well as number of cell cycle-related genes, which may or may not be directly linked to p53.
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Arsenic trioxide induces oxidative stress, DNA damage, and mitochondrial pathway of apoptosis in human leukemia (HL-60) cells.
J. Exp. Clin. Cancer Res.
PUBLISHED: 03-26-2014
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Acute promyelocytic leukemia (APL) is a subtype of acute myeloid leukemia (AML), which accounts for approximately 10% of all acute myloid leukemia cases. It is a blood cancer that is formed by chromosomal mutation. Each year in the United States, APL affects about 1,500 patients of all age groups and causes approximately 1.2% of cancer deaths. Arsenic trioxide (ATO) has been used successfully for treatment of APL patients, and both induction and consolidated therapy have resulted in complete remission. Recently published studies from our laboratory have demonstrated that ATO pharmacology as an anti-leukemic drug is associated with cytotoxic and genotoxic effects in leukemia cells.
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Diastolic function improves after resolution of takotsubo cardiomyopathy.
Clin Physiol Funct Imaging
PUBLISHED: 03-11-2014
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Takotsubo cardiomyopathy (TTC) is a condition of reversible left ventricular (LV) systolic dysfunction. However, the diastolic function (DF) manifestations of TTC have not been widely investigated. We performed a bicentric study with retrospective analysis of DF in patients with TTC, during onset and at follow-up.
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Evaluation of the Efficacy of Methylprednisolone, Etoricoxib and a Combination of the Two Substances to Attenuate Postoperative Pain and PONV in Patients Undergoing Laparoscopic Cholecystectomy: A Prospective, Randomized, Placebo-controlled Trial.
Korean J Pain
PUBLISHED: 03-07-2014
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Establishment of laparoscopic cholecystectomy as an outpatient procedure has accentuated the clinical importance of reducing early postoperative pain, as well as postoperative nausea and vomiting (PONV). We therefore planned to evaluate the role of a multimodal approach in attenuating these problems.
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A stress-responsive late embryogenesis abundant protein 7 (CsLEA7) of tea [Camellia sinensis (L.) O. Kuntze] encodes for a chaperone that imparts tolerance to Escherichia coli against stresses.
Mol. Biol. Rep.
PUBLISHED: 02-15-2014
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The present study characterized CsLEA7, a group 7 late embryogenesis abundant (LEA) gene, from tea [Camellia sinensis (L.) O. Kuntze]. The gene had an open reading frame of 462 base pairs encoding 153 amino acids with calculated molecular weight of 16.63 kDa and an isoelectric point (pI) of 4.93. Analysis revealed CsLEA7 to be an intrinsically ordered protein consisting of nine ?-strands and two ?-helices. CsLEA7 expressed ubiquitously in all the tissues analyzed with highest level of transcripts in mature leaf as compared to in flower bud, younger leaves, stem and fruit. Expression was the least in root tissue. CsLEA7 exhibited up-regulation in response to low temperature, polyethylene glycol-8000, sodium chloride and hydrogen peroxide in tea. Analysis of the promoter of CsLEA7 revealed a core promoter element and distinct cis-acting regulatory elements regulating gene expression under abiotic stresses. CsLEA7 exhibited chaperonic activity as evinced by protection to malate dehydrogenase against heat denaturation assay. Recombinant Escherichia coli cells producing CsLEA7 exhibited improved tolerance against diverse cues: polyethylene glycol-8000, sodium chloride, hydrogen peroxide and low temperature signifying its role in imparting stress tolerance.
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Changes in parathyroid proteome in patients with primary hyperparathyroidism due to sporadic parathyroid adenomas.
Clin. Endocrinol. (Oxf)
PUBLISHED: 02-13-2014
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The pathogenesis of parathyroid tumours is only partially understood. A direct approach using proteomics could be a promising tool to increase our understanding of parathyroid tumorigenesis. The aim of the study was to investigate differentially expressed proteins to explore the underlying molecular basis of the disease and identify potential target proteins responsible for the genesis of adenoma.
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RNA-seq-mediated transcriptome analysis of actively growing and winter dormant shoots identifies non-deciduous habit of evergreen tree tea during winters.
Sci Rep
PUBLISHED: 02-10-2014
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Tea [Camellia sinensis (L.) O. Kuntze] is a perennial tree which undergoes winter dormancy and unlike deciduous trees, the species does not shed its leaves during winters. The present work dissected the molecular processes operating in the leaves during the period of active growth and winter dormancy through transcriptome analysis to understand a long-standing question: why should tea be a non-deciduous species? Analyses of 24,700 unigenes obtained from 57,767 primarily assembled transcripts showed (i) operation of mechanisms of winter tolerance, (ii) down-regulation of genes involved in growth, development, protein synthesis and cell division, and (iii) inhibition of leaf abscission due to modulation of senescence related processes during winter dormancy in tea. These senescence related processes exhibited modulation to favour leaf abscission (i) in deciduous Populus tremula during winters, and (ii) also in tea but under osmotic stress during which leaves also abscise. These results validated the relevance of the identified senescence related processes for leaf abscission and suggested their operation when in need in tea.
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Comparison of dexmedetomidine, propofol and midazolam for short-term sedation in postoperatively mechanically ventilated neurosurgical patients.
J Clin Diagn Res
PUBLISHED: 02-08-2014
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Effective management of analgesia and sedation in the intensive care unit depends on the needs of the patient, subjective and/or objective measurement and drug titration to achieve specific endpoints.
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Effect of cold working on biocompatibility of Ni-free high nitrogen austenitic stainless steels using Dalton's Lymphoma cell line.
Mater Sci Eng C Mater Biol Appl
PUBLISHED: 01-14-2014
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The aims of the present work are to explore the effect of cold working on in-vitro biocompatibility of indigenized low cost Ni-free nitrogen containing austenitic stainless steels (HNSs) and to compare it with conventionally used biomedical grade, i.e. AISI 316L and 316LVM, using Dalton's Lymphoma (DL) cell line. The MTT assay [3-(4,5-dimethythiazol 2-yl)-2,5-diphenyltetrazolium bromide] was performed on DL cell line for cytotoxicity evaluation and cell adhesion test. As a result, it was observed that the HNS had higher cell proliferation and cell growth and it increases by increasing nitrogen content and degree of cold working. The surface wettability of the alloys was also investigated by water contact angle measurements. The value of contact angles was found to decrease with increase in nitrogen content and degree of cold working. This indicates that the hydrophilic character increases with increasing nitrogen content and degree of cold working which further attributed to enhance the surface free energy (SFE) which would be conducive to cell adhesion which in turn increases the cell proliferation.
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Bacterial diversity dynamics associated with different diets and different primer pairs in the rumen of kankrej cattle.
PLoS ONE
PUBLISHED: 01-01-2014
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The ruminal microbiome in herbivores plays a dominant role in the digestion of lignocellulose and has potential to improve animal productivity. Kankrej cattle, a popular native breed of the Indian subcontinent, were used to investigate the effect of different dietary treatments on the bacterial diversity in ruminal fractions using different primer pairs. Two groups of four cows were assigned to two primary diets of either dry or green forages. Each group was fed one of three dietary treatments for six weeks each. Dietary treatments were; K1 (50% dry/green roughage: 50% concentrate), K2 (75% dry/green roughage: 25% concentrate) and K3 (100% dry/green roughage). Rumen samples were collected using stomach tube at the end of each dietary period and separated into solid and liquid fractions. The DNA was extracted and amplified for V1-V3, V4-V5 and V6-V8 hypervariable regions using P1, P2 and P3 primer pairs, sequenced on a 454 Roche platform and analyzed using QIIME. Community compositions and the abundance of most bacterial lineages were driven by interactions between primer pair, dietary treatment and fraction. The most abundant bacterial phyla identified were Bacteroidetes and Firmicutes however, the abundance of these phyla varied between different primer pairs; in each primer pair the abundance was dependent on the dietary treatment and fraction. The abundance of Bacteroidetes in cattle receiving K1 treatment indicate their diverse functional capabilities in the digestion of both carbohydrate and protein while the predominance of Firmicutes in the K2 and K3 treatments signifies their metabolic role in fibre digestion. It is apparent that both liquid and solid fractions had distinct bacterial community patterns (P<0.001) congruent to changes in the dietary treatments. It can be concluded that the P1 primer pair flanking the V1-V3 hyper-variable region provided greater species richness and diversity of bacterial populations in the rumen of Kankrej cattle.
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Microenvironmental stiffness enhances glioma cell proliferation by stimulating epidermal growth factor receptor signaling.
PLoS ONE
PUBLISHED: 01-01-2014
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The aggressive and rapidly lethal brain tumor glioblastoma (GBM) is associated with profound tissue stiffening and genomic lesions in key members of the epidermal growth factor receptor (EGFR) pathway. Previous studies from our laboratory have shown that increasing microenvironmental stiffness in culture can strongly enhance glioma cell behaviors relevant to tumor progression, including proliferation, yet it has remained unclear whether stiffness and EGFR regulate proliferation through common or independent signaling mechanisms. Here we test the hypothesis that microenvironmental stiffness regulates cell cycle progression and proliferation in GBM tumor cells by altering EGFR-dependent signaling. We began by performing an unbiased reverse phase protein array screen, which revealed that stiffness modulates expression and phosphorylation of a broad range of signals relevant to proliferation, including members of the EGFR pathway. We subsequently found that culturing human GBM tumor cells on progressively stiffer culture substrates both dramatically increases proliferation and facilitates passage through the G1/S checkpoint of the cell cycle, consistent with an EGFR-dependent process. Western Blots showed that increasing microenvironmental stiffness enhances the expression and phosphorylation of EGFR and its downstream effector Akt. Pharmacological loss-of-function studies revealed that the stiffness-sensitivity of proliferation is strongly blunted by inhibition of EGFR, Akt, or PI3 kinase. Finally, we observed that stiffness strongly regulates EGFR clustering, with phosphorylated EGFR condensing into vinculin-positive focal adhesions on stiff substrates and dispersing as microenvironmental stiffness falls to physiological levels. Our findings collectively support a model in which tissue stiffening promotes GBM proliferation by spatially and biochemically amplifying EGFR signaling.
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Pseudoarthrosis and fracture: interaction between severe vitamin D deficiency and primary hyperparathyroidism.
Singapore Med J
PUBLISHED: 11-27-2013
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A young woman with severe vitamin D deficiency presented with proximal muscle weakness, fragility fracture and pseudoarthrosis. On evaluation, she was found to have hypercalcaemia, a single parathyroid adenoma and an undetectable 25-hydroxyvitamin D level. She received parenteral cholecalciferol and subsequently underwent curative parathyroidectomy. Postoperatively, she had hungry bone syndrome, which she gradually recovered from with calcium and calcitriol replacement. Notably, her calcium levels were in the lower limit of normal range and associated with elevated alkaline phosphatase levels at postoperative Day 14. Follow-up for the next four years showed that the patient had remarkable symptomatic and radiological improvements. In this report, we discuss the pathophysiological interactions between vitamin D deficiency and associated primary hyperparathyroidism.
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Chordoma of skull base presenting as nasopharyngeal mass.
J Neurosci Rural Pract
PUBLISHED: 11-01-2013
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While the nasopharynx is most commonly regarded by the otolaryngologist as a primary site of neoplastic involvement, it is also an avenue of spread of base-of-the-skull tumors presenting as bulging nasopharyngeal masses. Chordoma is a relatively rare tumor of the skull base and sacrum thought to originate from embryonic remnants of the notochord. Chordomas arising from the skull base/clivus are typically locally aggressive with lytic bone destruction. The optimal treatment may be photon/proton radiotherapy alone or combined with a gross total resection, when feasible. We report a case of intracranial chordoma presenting as nasopharyngeal mass.
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Development of a radiopharmaceutical dose calculator for pediatric patients undergoing diagnostic nuclear medicine studies.
Indian J Nucl Med
PUBLISHED: 10-29-2013
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It is important to ensure that as low as reasonably achievable (ALARA) concept during the radiopharmaceutical (RPH) dose administration in pediatric patients. Several methods have been suggested over the years for the calculation of individualized RPH dose, sometimes requiring complex calculations and large variability exists for administered dose in children. The aim of the present study was to develop a software application that can calculate and store RPH dose along with patient record.
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Identification of crucial amino acids of bacterial Peptide deformylases affecting enzymatic activity in response to oxidative stress.
J. Bacteriol.
PUBLISHED: 10-18-2013
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Peptide deformylase (PDF) is an essential bacterial metalloprotease involved in deformylation of N-formyl group from nascent polypeptide chains during protein synthesis. Iron-containing variants of this enzyme from Salmonella enterica serovar Typhimurium (sPDF) and Mycobacterium tuberculosis (mPDF), although inhibited by oxidizing agents like H2O2, exhibited strikingly different 50% inhibitory concentrations (IC50s) that ranged from nanomolar (sPDF) to millimolar (mPDF) levels. Furthermore, the metal dissociation rate was higher in sPDF than mPDF. We hypothesized that a restriction in entry of environmental oxygen or oxidizing agents into the active site of mPDF might be the cause for such discrepancies between two enzymes. Since the active-site residues of the two proteins are similar, we evaluated the role of the oxidation-prone noncatalytic residue(s) in the process. Amino acid sequence analysis revealed that Cys-130 of sPDF corresponds to Met-145 of mPDF and that they are away from the active sites. Swapping methionine with cysteine in mPDF, the M145C protein displayed a drastic decrease in the IC50 for H2O2 and an increased metal dissociation rate compared to the wild type. Matrix-assisted laser desorption ionization (MALDI) analysis of a trypsin-digested fragment containing Cys-145 of the M145C protein also indicated its increased susceptibility to oxidation. To incorporate residues identical to those of mPDF, we created a double mutant of sPDF (C130M-V63C) that showed increased IC50 for H2O2 compared to the wild type. Interestingly, the oxidation state of cysteines in C130M-V63C was unaffected during H2O2 treatment. Taken together, our results unambiguously established the critical role of noncatalytic cysteine/methionine for enzymatic sensitivity to H2O2 and, thus, for conferring behavioral distinction of bacterial PDFs under oxidative stress conditions.
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HSF1-mediated regulation of tumor cell apoptosis: a novel target for cancer therapeutics.
Future Oncol
PUBLISHED: 10-11-2013
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Programmed cell death/apoptosis is a genetically conserved phenomenon involved in many biological processes including reconstruction of multicellular organisms and elimination of old or damaged cells. It is regulated by the activation/deactivation of PKC in response to exogenous and endogenous stimuli. PKC is activated under stress by a series of downstream signaling cascades, which ultimately induce HSF1 activation, which results in overexpression of heat shock proteins. Overexpression of heat shock proteins interferes in the apoptotic pathway, while their blocking results in apoptosis. Therefore, HSF1 could be a novel therapeutic target against a variety of tumors. Several pharmacological inhibitors of PKC have been demonstrated to exert inhibitory effects on the activation of HSF1 and, therefore, induce apoptosis in tumor cells. However, studies regarding the role of pharmacological inhibitors in the regulation of apoptosis and possible anti-tumor therapeutic intervention are still unknown or in their infancy. Therefore, an attempt has been made to delineate the precise role of HSF1 in the regulation of apoptosis and its prospects in cancer therapeutics.
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The therapeutic potential of GPR120: a patent review.
Expert Opin Ther Pat
PUBLISHED: 10-06-2013
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Introduction: Type 2 diabetes (T2D) is a complex metabolic disorder characterized by persistent hyperglycemia and a wide range of underlying metabolic defects. The prevalence and incidence of T2D are expected to dramatically increase in the near-future and consequently, there is a significant medical need for diabetes care. Many targets are under investigation to lower the plasma glucose levels or increase the insulin sensitivity. Despite newer drug classes emerging as viable long-term treatment options for the management of T2D, achieving an optimal glycemic control along with sufficient effectiveness over the course of the disease remains a challenge. In this regard, among several G-protein-coupled receptors (GPCRs), GPR120 and GPR40 have recently been considered as viable targets for diabetes and metabolic disorders. Areas covered: This article reviews the current literature on the discovery and development of GPR120 agonists in diabetes and metabolic disorders and updates on the published patents in this field. The patent study for this review has been carried out using multiple electronic databases including SciFinder and Thomson Reuters Integrity. Expert opinion: A paradigm shift in the treatment of diabetes is needed, wherein a single therapeutic agent could target diabetes and its associated disorders such as high plasma glucose level and inflammation, with excellent safety and tolerability profile. In this regard, agonists of GPR120 or dual-agonist GPR120 and GPR40 are highly anticipated as therapeutic approaches for the treatment of diabetes on the basis of their novel glucose-dependent mechanism of action.
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A fluorescent peroxidase probe increases the sensitivity of commercial ELISAs by two orders of magnitude.
Chem. Commun. (Camb.)
PUBLISHED: 09-27-2013
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The low detection sensitivity of enzyme linked immunosorbent assays (ELISAs) is a central problem in science and limits progress in multiple areas of biology and medicine. In this report we demonstrate that the hydrocyanines, a family of fluorescent reactive oxygen species (ROS) probes, can act as turn on fluorescent horseradish peroxidase (HRP) probes and thereby increase the sensitivity of conventional ELISAs by two orders of magnitude.
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Biosynthesis of silver nanoparticles: Elucidation of prospective mechanism and therapeutic potential.
J Colloid Interface Sci
PUBLISHED: 08-27-2013
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The synthesis of silver nanoparticles (AgNPs) was accomplished using Syzygium cumini fruit extract at room temperature. Various techniques were used to characterize the newly synthesized silver nanoparticles and their size was determined to be 10-15nm. Important findings of this study were the identification of biomolecules responsible for the synthesis of silver nanoparticles and elucidate the mechanism of biosynthesis. Flavonoids present in S. cumini were mainly responsible for the reduction and the stabilization of nanoparticles. The antioxidant properties of AgNPs were evaluated using various assays. The nanoparticles were also found to destroy Dalton lymphoma cell lines under in vitro condition. Silver nanoparticles (100?g/mL) decreased the viability of Dalton lymphoma (DL) cell lines up to 50%. The studies describing the biosynthesis of silver nanoparticles by fruit extract followed by the investigation of synthesis mechanism and anti-cancer activities may be useful for nanobiotechnology research opening a new arena in this field.
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The GATA and SORLIP motifs in the 3-hydroxy-3-methylglutaryl-CoA reductase promoter of Picrorhiza kurrooa for the control of light-mediated expression.
Funct. Integr. Genomics
PUBLISHED: 08-19-2013
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Light upregulates the expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) in Picrorhiza kurrooa, an endangered medicinal herb. Upstream sequences of HMGR of P. kurrooa (PropkHMGR) were analyzed in relation to its role in light-mediated regulation of gene expression. GATA motif in PropkHMGR exhibited stronger DNA-protein interaction with the nuclear extract of dark-exposed plants in contrast to SORLIP that exhibited stronger binding with the nuclear extract of light-exposed plants. Analysis of PropkHMGR (PropkHMGR-D1, -1,059/-1) and its deletion fragments PropkHMGR-D2 (-825/-1), PropkHMGR-D3 (-651/-1), PropkHMGR-D4 (-452/-1), and PropkHMGR-D5 (-101/-1) in Arabidopsis thaliana showed PropkHMGR to regulate gene expression [?-glucuronidase (GUS) was used as a reporter gene] at all the developmental stages but only in actively dividing tissues, excluding anthers. Whereas, PropkHMGR-D2 regulated GUS expression in relatively older seedlings but the expression was observed only in shoot apical meristem, root tips, and anthers. PropkHMGR-mediated gene expression was higher in dark as compared to that in the light in Arabidopsis across four temperatures studied. As opposed to the results in P. kurrooa, GATA motifs exhibited DNA-protein interaction with nuclear extract of light-exposed plants of Arabidopsis. SORLIP motifs in Arabidopsis also exhibited DNA-protein interaction with nuclear extract of light-exposed plants as in P. kurrooa. Data showed that (1) PropkHMGR regulated light-mediated gene expression and (2) GATA motif exhibited an inverse relationship between strength of DNA-protein interaction and the gene expression whereas the relationship was species specific for SORLIP.
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Proteomic Identification of Syzygium cumini Seed Extracts by MALDI-TOF/MS.
Appl. Biochem. Biotechnol.
PUBLISHED: 08-02-2013
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Syzygium cumini is traditionally used medicinal plant. The different part of the plant such as bark, leaves, seed and fruits are widely used as an alternative medicine in various diseases. Although the scientific community has a strong interest on S. cumini seed biochemistry focusing on metabolite composition, proteins have not yet been investigated. In the present study, we have applied a proteomic approach to study the proteome of the S. cumini seed using phenol extraction method for protein isolation, which were never analysed before. Fifteen brightly silver stained protein spots were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry after resolving on two-dimensional gel electrophoresis. These proteins have been found to involve in various functions such as antifungal, sulphur metabolism, carbohydrate metabolism, fruit ripening and softening, dormancy breaking and seed germination, hormone signalling, secondary metabolite transport, defence and stress response, nitrogen metabolism, synthesis and stabilization. Amongst the identified protein, lactoferrin was a mammalian origin protein with high nutritious and pharmaceutical value, which was purified by different types of chromatographic techniques and confirmed by western blotting. The antibacterial activity of lactoferrin was assessed by disc diffusion assay. We suggest that the protein constituents of S. cumini may have role in various functions required for plant physiology and its dietary values.
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Methylation status of the CpG islands in vitamin D and calcium-sensing receptor gene promoters does not explain the reduced gene expressions in parathyroid adenomas.
J. Clin. Endocrinol. Metab.
PUBLISHED: 08-02-2013
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The exact mechanism causing decreased expression of the vitamin D receptor (VDR) and calcium-sensing receptor (CASR) genes in parathyroid adenoma is not known, but methylation of promoter regions is often detected during epigenetic downregulation of gene expression. We investigated whether epigenetic silencing is involved in the decreased expression of VDR and CASR.
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Heparin binding carboxypeptidase E protein exhibits antibacterial activity in human semen.
Int. J. Biol. Macromol.
PUBLISHED: 07-31-2013
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Carboxypeptidase E (CPE) cleaves basic amino acid residues at the C-terminal end and involves in the biosynthesis of numerous peptide hormones and neurotransmitters. It was purified from human seminal plasma by ion exchange, heparin affinity and gel filtration chromatography followed by identification through SDS-PAGE and MALDI-TOF/MS analysis, which was further confirmed by western blotting. CPE was characterized as glycoprotein by Periodic Acid Schiff (PAS) staining and treating with deglycosylating enzyme N-glycosidase F. The interaction of CPE with heparin was illustrated by surface plasmon resonance (SPR) and in silico interaction analysis. The association constant (KA) and dissociation constant (KD) of CPE with heparin was determined by SPR and found to be 1.06×10(5)M and 9.46×10(-6)M, respectively. It was detected in human spermatozoa also by western blotting using mouse anti-CPE primary antibody. 20-100?g/ml concentration of CPE was observed as highly effective in killing Escherichia coli by colony forming unit (CFU) assay. We suggest that CPE might act not only in the innate immunity of male reproductive tract but also regulate sperm fertilization process by interacting heparin.
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A giant saphenous vein graft aneurysm causing left internal mammary artery to left anterior descending artery graft compression.
Am. J. Cardiol.
PUBLISHED: 07-16-2013
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Saphenous vein graft aneurysm is a rare complication after coronary artery bypass surgery. It is defined as a localized dilatation of the vessel to 1.5× the expected normal diameter. We report a case of 67-year-old man who presented with angina 23 years after coronary artery bypass graft. He was found to have a giant saphenous vein graft aneurysm to diagonal artery compressing the left internal mammary artery graft to the left anterior descending artery. The patient underwent aneurysm resection with satisfactory outcome.
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New aspects and strategies for methane mitigation from ruminants.
Appl. Microbiol. Biotechnol.
PUBLISHED: 07-12-2013
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The growing demand for sustainable animal production is compelling researchers to explore the potential approaches to reduce emissions of greenhouse gases from livestock that are mainly produced by enteric fermentation. Some potential solutions, for instance, the use of chemical inhibitors to reduce methanogenesis, are not feasible in routine use due to their toxicity to ruminants, inhibition of efficient rumen function or other transitory effects. Strategies, such as use of plant secondary metabolites and dietary manipulations have emerged to reduce the methane emission, but these still require extensive research before these can be recommended and deployed in the livestock industry sector. Furthermore, immunization vaccines for methanogens and phages are also under investigation for mitigation of enteric methanogenesis. The increasing knowledge of methanogenic diversity in rumen, DNA sequencing technologies and bioinformatics have paved the way for chemogenomic strategies by targeting methane producers. Chemogenomics will help in finding target enzymes and proteins, which will further assist in the screening of natural as well chemical inhibitors. The construction of a methanogenic gene catalogue through these approaches is an attainable objective. This will lead to understand the microbiome function, its relation with the host and feeds, and therefore, will form the basis of practically viable and eco-friendly methane mitigation approaches, while improving the ruminant productivity.
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Adeno-associated virus (AAV) vectors in gene therapy: immune challenges and strategies to circumvent them.
Rev. Med. Virol.
PUBLISHED: 06-04-2013
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AAV-based gene transfer protocols have shown remarkable success when directed to immune-privileged sites such as for retinal disorders like Lebers congenital amaurosis. In contrast, AAV-mediated gene transfer into liver or muscle tissue for diseases such as hemophilia B, ?1 anti-trypsin deficiency and muscular dystrophy has demonstrated a decline in gene transfer efficacy over time. It is now known that in humans, AAV triggers specific pathways that recruit immune sensors. These factors initiate an immediate reaction against either the viral capsid or the vector encoded protein as part of innate immune response or to produce a more specific adaptive response that generates immunological memory. The vector-transduced cells are then rapidly destroyed due to this immune activation. However, unlike other viral vectors, AAV is not immunogenic in murine models. Its immunogenicity becomes apparent only in large animal models and human subjects. Moreover, humans are natural hosts to AAV and exhibit a high seroprevalence against AAV vectors. This limits the widespread application of AAV vectors into patients with pre-existing neutralising antibodies or memory T cells. To address these issues, various strategies are being tested. Alternate serotype vectors (AAV1-10), efficient expression cassettes, specific tissue targeting, immune-suppression and engineered capsid variants are some approaches proposed to minimise this immune stimulation. In this review, we have summarised the nature of the immune response documented against AAV in various pre-clinical and clinical settings and have further discussed the strategies to evade them.
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Osmotin-expressing transgenic tea plants have improved stress tolerance and are of higher quality.
Transgenic Res.
PUBLISHED: 06-02-2013
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Drought is a major stress that affects the yield and quality of tea, a widely consumed beverage crop grown in more than 20 countries of the world. Therefore, osmotin gene-expressing transgenic tea plants produced using earlier optimized conditions were evaluated for their tolerance of drought stress and their quality. Improved tolerance of polyethylene glycol-induced water stress and faster recovery from stress were evident in transgenic lines compared with the normal phenotype. Significant improvements in growth under in-vitro conditions were also observed. Besides enhanced reactive oxygen species-scavenging enzyme activity, the transgenic lines contained significantly higher levels of flavan-3-ols and caffeine, key compounds that govern quality and commercial yield of the beverage. The selected transgenic lines have the potential to meet the demands of the tea industry for stress-tolerant plants with higher yield and quality. These traits of the transgenic lines can be effectively maintained for generations because tea is commercially cultivated through vegetative propagation only.
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Simultaneous presence of both open metal sites and free functional organic sites in a noncentrosymmetric dynamic metal-organic framework with bimodal catalytic and sensing activities.
Chemistry
PUBLISHED: 05-31-2013
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Assimilation of open metal sites (OMSs) and free functional organic sites (FOSs) with a framework strut has opened up a new route for the fabrication of novel metal-organic materials, thereby providing a unique opportunity to explore their multiple functionalities. A new metal-organic framework (MOF), {[Cu(ina)2 (H2 O)][Cu(ina)2 (bipy)]?2?H2 O}n (1) (ina=isonicotinate, bipy=4,4-bipyridine), has been synthesized and characterized. Complex 1 is crystallized in the orthorhombic noncentrosymmetric space group Aba2 and consists of two different 2D coordination polymers, [Cu(ina)2 (H2 O)]n and [Cu(ina)2 (bipy)]n , with entrapped solvent water molecules. Hydrogen-bonding interactions assemble these two different 2D coordination layers in a single-crystal structure with interdigitation of pendant 4,4-bipy from one layer into the groove of another. Upon removal of guest molecules, 1 undergoes a structural transformation in single-crystal-to-single-crystal fashion with expansion of the effective void space. Each metal center is five-coordinated and thus can potentially behave as an OMS, and the free pyridyl groups of pendant 4,4-bipy moieties and free ?C?O groups can act as free FOSs. Thus, owing to presence of both OMSs and free FOSs, the framework exhibits multifunctional properties. Owing to the presence of OMSs, the framework can act as a Lewis acid catalyst as well as a small-molecule sensor material, and in a similar way, owing to the presence of free FOSs, it performs as a Lewis base catalyst and a cation sensor material. Furthermore, owing to noncentrosymmetry with large polarity along a particular direction, it shows strong second-harmonic generation/nonlinear optical (SHG-NLO) activity.
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Discovery of P3971 an orally efficacious novel anticancer agent targeting HIF-1? and STAT3 pathways.
Anticancer Agents Med Chem
PUBLISHED: 05-22-2013
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Hypoxia-inducible factor-1? (HIF-1?) and signal transducer and activator of transcription 3 (STAT3) are transcription factors and are activated in response to hypoxia. Both HIF-1? and STAT3 regulate various aspects of cancer biology such as cell survival, proliferation, angiogenesis etc. and are constitutively expressed in a wide range of human cancers. In the last decade, over expression of HIF-1? and STAT3 has been demonstrated in many common human cancers, thereby emerging as highly compelling anticancer targets for drug discovery. We herein report the design and synthesis of new imidazopyridine based potent dual inhibitors of HIF-1? and STAT3 pathways. The lead compound of this series P3971 has been identified as a potent inhibitor of HIF-1? (200 nM) and STAT3 (350 nM) with significant antiproliferative activity against various cancer cell lines. Moreover, P3971 was also found to be orally efficacious in HCT116 (colon cancer) and H460 (lung cancer) xenograft mice models.
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Development of EMA-2 recombinant antigen based enzyme-linked immunosorbent assay for seroprevalence studies of Theileria equi infection in Indian equine population.
Vet. Parasitol.
PUBLISHED: 05-16-2013
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Equine piroplasmosis is a tick-transmitted protozoan disease caused by Theileria equi and/or Babesia caballi. In the present study, we expressed a 53kDa protein from the truncated EMA-2 gene of T. equi (Indian strain) and developed EMA-2ELISA using this expressed protein. This ELISA is able to detect T. equi-specific antibodies in experimentally infected animals as early as 9 days post-infection. The assay developed was validated with the OIE recommended competitive ELISA (cELISA) on 120 serum samples and significant agreement (kappa=0.93) was observed between results of both the ELISAs which indicates suitability of EMA-2ELISA for use in sero-diagnosis. Diagnostic sensitivity and specificity of EMA-2ELISA - as compared with cELISA - were 0.97 and 0.96, respectively. Analysis of 5651 equine serum samples - collected during 2007-2012 from 12 states of India representing eight agro-climatic zones - by EMA-2ELISA revealed 32.65% seroprevalence of T. equi in India. In conclusion, the EMA-2ELISA developed using the T. equi EMA-2 recombinant protein as antigen for detecting T. equi-specific antibodies has good diagnostic potential for sero-epidemiological surveys.
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Camurati-Engelmann Disease (Progressive Diaphyseal Dysplasia): Reports of an Indian Kindred.
Calcif. Tissue Int.
PUBLISHED: 01-16-2013
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Camurati-Engelmann disease (CED, OMIM 131300), or progressive diaphyseal dysplasia, is a rare autosomal dominant skeletal dysplasia, caused by mutations in the transforming growth factor-?1 (TGF?1) gene. We describe the first Indian CED family with genetic confirmation and presenting manifestations. The proband is a 17-year-old woman who presented with lower limb pain and proximal muscle weakness. Skeletal radiographs of the long bones revealed cortical, periosteal, and endosteal thickenings, predominantly affecting the diaphyses of the long bones. On detailed evaluation, there was a strong family history of bone disorder with similar symptoms of pain and radiological findings in several family members. Exon sequencing of the TGF?1 gene was performed in available family members. Based on clinical and radiographic studies and its familial nature, a diagnosis of CED was made and confirmed by mutation analysis. A heterozygous G to A transition in exon 4 of the TGF?1 gene (R218H) was detected in 5 out of 10 available family members, including 4 affecteds and 1 asymptomatic individual. Many of our affected individuals responded to glucocorticoids and cortical windowing. CED is a rare genetic disease with variable clinical manifestations and incomplete penetrance. CED needs to be considered in the differential diagnosis of nonspecific limb pain and waddling gait in all young individuals.
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A comparison between intensive and conventional cabergoline treatment of newly diagnosed patients with macroprolactinoma.
Clin. Endocrinol. (Oxf)
PUBLISHED: 01-07-2013
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Intensive treatment with cabergoline may lead to earlier reduction in prolactin and tumour volume in comparison to conventional schedule.
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Attending to the possibilities of action.
Philos. Trans. R. Soc. Lond., B, Biol. Sci.
PUBLISHED: 01-01-2013
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Actions taking place in the environment are critical for our survival. We review evidence on attention to action, drawing on sets of converging evidence from neuropsychological patients through to studies of the time course and neural locus of action-based cueing of attention in normal observers. We show that the presence of action relations between stimuli helps reduce visual extinction in patients with limited attention to the contralesional side of space, while the first saccades made by normal observers and early perceptual and attentional responses measured using electroencephalography/event-related potentials are modulated by preparation of action and by seeing objects being grasped correctly or incorrectly for action. With both normal observers and patients, there is evidence for two components to these effects based on both visual perceptual and motor-based responses. While the perceptual responses reflect factors such as the visual familiarity of the action-related information, the motor response component is determined by factors such as the alignment of the objects with the observers effectors and not by the visual familiarity of the stimuli. In addition to this, we suggest that action relations between stimuli can be coded pre-attentively, in the absence of attention to the stimulus, and action relations cue perceptual and motor responses rapidly and automatically. At present, formal theories of visual attention are not set up to account for these action-related effects; we suggest ways that theories could be expected to enable action effects to be incorporated.
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Acrogigantism and facial asymmetry: McCune-Albright syndrome.
J. Pediatr. Endocrinol. Metab.
PUBLISHED: 12-08-2011
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McCune-Albright syndrome (MAS) is characterized by a triad of poly/monoostotic fibrous dysplasia, café-au-lait macules and hyperfunctioning endocrinopathies. Association of MAS with GH excess is rare, and in most of the instances somatotropinoma has not been documented. Treatment of patients of MAS with acromegaly is difficult because of thickened calvarium and dysplastic skull bone. We report a 17-year-old girl, who presented with cranio-facial fibrous dysplasia, café-au-lait macules and also had acromegaly due to pituitary macroadenoma, and treated with gamma knife radiosurgery.
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Contributions of talin-1 to glioma cell-matrix tensional homeostasis.
J R Soc Interface
PUBLISHED: 12-07-2011
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The ability of cells to adapt their mechanical properties to those of the surrounding microenvironment (tensional homeostasis) has been implicated in the progression of a variety of solid tumours, including the brain tumour glioblastoma multiforme (GBM). GBM tumour cells are highly sensitive to extracellular matrix (ECM) stiffness and overexpress a variety of focal adhesion proteins, such as talin. While talin has been shown to play critical early roles in integrin-based force-sensing in non-tumour cells, it remains unclear whether this protein contributes to tensional homeostasis in GBM cells. Here, we investigate the role of the talin isoform talin-1 in enabling human GBM cells to adapt to ECM stiffness. We show that human GBM cells express talin-1, and we use RNA interference to suppress talin-1 expression without affecting levels of talin-2, vinculin or phosphorylated focal adhesion kinase. Knockdown of talin-1 strongly reduces both cell spreading area and random migration speed but does not significantly affect overall focal adhesion size distributions. Most strikingly, atomic force microscopy indentation reveals that talin-1 suppression compromises adaptation of cell stiffness to changes in ECM stiffness. Together, these data support a role for talin-1 in the maintenance of tensional homeostasis in GBM and suggest a functional role for enriched talin expression in this tumour.
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Human serum albumin as a new interacting partner of prolactin inducible protein in human seminal plasma.
Int. J. Biol. Macromol.
PUBLISHED: 11-23-2011
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Prolactin inducible protein (PIP) is a 17 kDa glycoprotein. It binds to many proteins including fibrinogen, actin, keratin, myosin, immunoglobulin G, CD4, and human zinc-alpha-2 glycoprotein. Its ability to bind a large array of proteins indicates its multifaceted role in various biological processes, such as fertility, immunoregulation, antimicrobial activity, apoptosis, and tumor progression. Here, we present the first report of native human serum albumin (HSA)-PIP complex formation in seminal plasma. The complex was purified by chromatographic separation techniques, analyzed by gel electrophoresis, identified by MALDI-TOF mass spectrometry and validated by co-immunoprecipitation coupled with western blotting experiments. Moreover, the behavior of complex in solution was analyzed by dynamic light scattering and interacting residues were identified by in silico protein-protein docking. The purified protein complex shows two bands (67 kDa and 17 kDa) on SDS-PAGE gel and a single band (~85 kDa) on native PAGE gel. The predicted complex structure has 13 intermolecular hydrogen bonds, which may contribute to the overall stability of the complex. As HSA has been known to preserve the motility of sperm, native HSA-PIP complex formation may point towards an important role of PIP, which can directly be correlated with male fertility/infertility.
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900-MHz microwave radiation promotes oxidation in rat brain.
Electromagn Biol Med
PUBLISHED: 11-04-2011
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Recently, there have been several reports referring to detrimental effects due to radio frequency electromagnetic fields (RF-EMF) exposure. Special attention was given to investigate the effect of mobile phone exposure on the rat brain. Since the integrative mechanism of the entire body lies in the brain, it is suggestive to analyze its biochemical aspects. For this, 35-day old Wistar rats were exposed to a mobile phone for 2 h per day for a duration of 45 days where specific absorption rate (SAR) was 0.9 W/Kg. Animals were divided in two groups: sham exposed (n = 6) and exposed group (n = 6). Our observations indicate a significant decrease (P < 0.05) in the level of glutathione peroxidase, superoxide dismutase, and an increase in catalase activity. Moreover, protein kinase shows a significant decrease in exposed group (P < 0.05) of hippocampus and whole brain. Also, a significant decrease (P < 0.05) in the level of pineal melatonin and a significant increase (P < 0.05) in creatine kinase and caspase 3 was observed in exposed group of whole brain as compared with sham exposed. Finally, a significant increase in the level of ROS (reactive oxygen species) (P < 0.05) was also recorded. The study concludes that a reduction or an increase in antioxidative enzyme activities, protein kinase C, melatonin, caspase 3, and creatine kinase are related to overproduction of reactive oxygen species (ROS) in animals under mobile phone radiation exposure. Our findings on these biomarkers are clear indications of possible health implications.
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Primary adenocarcinoma of the frontal sinus.
Ear Nose Throat J
PUBLISHED: 10-29-2011
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When tumor involvement of the frontal sinus occurs, it is usually the result of the direct spread of the mass from the nasal cavity or anterior ethmoid sinuses. Primary frontal sinus tumors are extremely rare. We describe a case of primary adenocarcinoma of the frontal sinus in a 53-year-old man. The patient refused surgery, so he was treated with chemotherapy followed by radiotherapy. At the 1-year follow-up, only a slight reduction of the swelling was noted. Thereafter, he was lost to follow-up. This case represents a typical example of a paranasal cancer extending beyond the bony margins, with a silent onset that simulated benign disease. The best hope for an early diagnosis of such a tumor lies in the greater use of computed tomography to assess chronic rhinosinusitis.
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What is Visualize?

JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

How does it work?

We use abstracts found on PubMed and match them to JoVE videos to create a list of 10 to 30 related methods videos.

Video X seems to be unrelated to Abstract Y...

In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.