Search

Macromolecular complexes are essential to conserved biological processes, but their prevalence across animals is unclear. By combining extensive biochemical fractionation with quantitative mass spectrometry, here we directly examined the composition of soluble multiprotein complexes among diverse metazoan models. Using an integrative approach, we generated a draft conservation map consisting of more than one million putative high-confidence co-complex interactions for species with fully sequenced genomes that encompasses functional modules present broadly across all extant animals. Clustering reveals a spectrum of conservation, ranging from ancient eukaryotic assemblies that have probably served cellular housekeeping roles for at least one billion years, ancestral complexes that have accrued contemporary components, and rarer metazoan innovations linked to multicellularity. We validated these projections by independent co-fractionation experiments in evolutionarily distant species, affinity purification and functional analyses. The comprehensiveness, centrality and modularity of these reconstructed interactomes reflect their fundamental mechanistic importance and adaptive value to animal cell systems., Introduction Elucidating the components, conservation and functions of multiprotein complexes is essential to understand cellular processes (1,2), but mapping physical association networks on a proteome-wide scale is challenging. The development [...]

In India, the cashew nut shelling (CNS) is an important task in all small-scale cashew nut processing industries. Workers engaged in CNS task commonly operate pedal-operated cashew nut sheller (CNSR) in awkward standing posture that can contribute to work-related musculoskeletal disorders (MSDs). However, investigations from an ergonomics perspective have barely been conducted in Indian small-scale cashew nut processing industries. Therefore, the present study aims to evaluate the postural workload in CNS task to propose an ergonomic design intervention in small-scale cashew nut processing industries. A combination of methods, including posture recording, postural assessment, interviews, questionnaire study, virtual prototyping, and digital human modeling and simulation, were used. Using the pedal-operated CNSR resulted in high-risk postures of MSDs. Based on these preliminary findings, a simple, improved, and adjustable hand-operated CNSR was designed and evaluated ergonomically. It was found that newly designed CNSR improved postures noticeably and provided comfort for workers. [ABSTRACT FROM AUTHOR]

Colour of light modulates the appearance of displayed artifacts in exhibition. To understand the effect of light on appearance of paintings, few studies have experimented to establish a relation between pleasantness with the correlated colour temperature (CCT) of light. However, some studies have found that CCT has no significant effect on pleasant appearance of painting. Therefore, in this article, an experiment with a qualitative (questionnaire on semantic differential scale, N = 30) approach was designed to understand the effect of CCT on appearance of paintings exhibited under LED lights. The findings from the experimental result depict that the appearance of paintings changes due to different CCTs of LEDs having the same illuminance. In addition, the result reveals that for both mediums of paintings considered in this study, in comparison to warm white LED and artificial daylight LED, cool white LED has appeared to be more pleasant having moderately warm feelings to the viewers. [ABSTRACT FROM AUTHOR]

Summary Cerebral cavernous malformations (CCMs) are vascular defects of the CNS that arise from loss of integrity of the endothelial cells lining blood capillaries, causing leakage of blood into the brain [ 1 ]. This results in headaches, seizures, and/or hemorrhagic stroke, depending on the location of the lesion. CCM affects 0.5% of the population and follows an autosomal dominant inheritance pattern caused by mutations in one of the three genes: CCM1 (gene name KRIT1 ), CCM2 (also known as malcavernin or OSM ), and CCM3 (gene name PDCD10 ) [ 2, 3 ], with the earliest onset and most severe prognosis occurring in CCM3 patients [ 4 ]. The three CCM genes encode structurally distinct scaffold proteins that function in multiple complexes [ 5–9 ]. Using the C. elegans germline as a model of multicellular tube development, we show here that CCM-3 is enriched at the luminal membrane of the germline and the contractile ring of dividing cells in the embryo. Loss of ccm-3 results in defective RAB-11-mediated endocytic recycling, which in turn is necessary for gonadal lumen (rachis) formation, completion of cytokinesis, and localization of cell-surface receptors. CCM-3-mediated localization of anillin and non-muscle myosin to the lateral surfaces of germ cells is required for proper cytoskeletal organization, subsequent oocyte growth, and localization of polarity proteins. Biochemical analysis reveals conservation of the STRIPAK complex and distinct roles for GCK-1 (germinal center kinase III family protein) and striatin/CASH-1 in controlling the localization and function of CCM-3. Taken together, our data establish CCM-3 as a novel regulator of rachis lumenization and polarity establishment during embryogenesis. [ABSTRACT FROM AUTHOR]

Medical Tourism is a concept that encompasses and adjoins two types of service sector viz. Medical treatment along with Tourism to various destinations of the host country. Medical tourism is the act of travelling to another nation for availing treatment of international standards at a cost which is otherwise very costly while having leisure tours to various attractive sites of the host country at the same time. Medical tourism is a sector which has not yet been explored to its optimum level in India. Kolkata or Calcutta, the capital of West Bengal, popularly known as 'The City of Joy' being remembered for its rich cultural heritage, is an important commercial, cultural, and educational centre of East India. Considering its proximity to Bangladesh, Nepal, and Bhutan as well as making its mark in Top 10 States of receiving foreign and domestic tourists, the pertinent study presents the scope and opportunities of Medical Tourism in Kolkata. The authors present a comparative analysis between Kolkata and other metropolitan cities with respect to different parameters like Foreign and Domestic Tourist arrivals (FTAs and DTAs), number of hospitals, number of Joint Commission International (JCI) accredited hospitals, number of National Accreditation Board for Hospitals & Health Care Providers(NABH) accredited hospitals, number of blood banks, cost of living and so on. Based on various literatures available, this paper focuses on identifying the scope and opportunities of medical tourism in Kolkata through SWOT (Strength, Weakness, Opportunity, Threat) analysis. [ABSTRACT FROM AUTHOR]

Though e-commerce and online shopping concepts are almost a couple of decades old, it is the advancement of information and communication technology and globalization, that has accelerated the shift of a large share of customer base from brick and mortar to its electronic counterpart. Various studies and statistics show that people are spending more time in different online activities – both financial and nonfinancial. The convenience of online services, especially of e-commerce has attracted consumers towards adapting this newer way of shopping. The paper highlights the major online activities performed by consumers. It also focuses on different factors affecting adoption of online shopping. [ABSTRACT FROM AUTHOR]

Significance: Reactive oxygen species (ROS) are produced as natural byproducts of metabolism and respiration. While physiological levels of ROS are required for vital cellular functions (e.g., development and proliferation), a living organism is faced with constant challenges due to accumulation or overproduction of ROS throughout its life. The life cycle of Leishmania parasite has led it to confront the highly oxidizing environment in the macrophage phagosomes, necessitating ROS homeostasis and signaling as key strategies for successful survival and pathogenicity. Recent Advances: Ascorbate peroxidase from Leishmania major (LmAPX) is the only heme peroxidase identified so far in Leishmania. Structural analysis and functional characterization of LmAPX have yielded interesting and novel insight on this enzyme. The protein has been found to be a hybrid of cytochrome c peroxidase and ascorbate peroxidase. This enzyme is colocalized with cytochrome c in the inner mitochondrial membrane facing the intermembrane space and shows higher activity toward cytochrome c oxidation. Critical Issues: Overexpression of LmAPX in L. major cells confers tolerance to oxidative stress-mediated cardiolipin oxidation and consequently protects cells from extensive protein damage. LmAPX-/- mutants show higher intracellular hydrogen peroxide (H2O2), which might signal for cellular transformation from noninfective procyclic to infective metacyclic form and ultimately apoptosis. Future Directions: Manipulation of LmAPX expression has significantly added to the present understanding of the parasite's defense network against oxidative damage caused by H2O2. The future investigations will address more exactly the signaling pathways involved in redox homeostasis. [ABSTRACT FROM AUTHOR]

The paper focuses on the behavioural aspect of undergraduate students who want to pursue higher education especially professional courses. The paper deals with how students perceive full time and part time courses along with course fee structure. Also the paper shows the perception of students regarding job guarantee along with course duration. The paper gives a clear idea that job guarantee is an important factor while going for professional course irrespective of gender. Not only the paper highlights important factors while selecting a professional course but also focuses on their perception towards migrating to other cities for fulfilling their dreams. The paper also comes up with the various opinions towards different sectors like FMCG/CD, Retail, Manufacturing, IT, and BPO/KPO and in which order they are preferred most [ABSTRACT FROM AUTHOR]

Abstract: The endogenous cation in peroxidases may contribute to the type of heme coordination. Here a series of ferric and ferrous derivatives of wild-type Leishmania major peroxidase (LmP) and of engineered K+ site mutants of LmP, lacking potassium cation binding site, has been examined by electronic absorption spectroscopy at 25°C. Using UV–visible spectrophotometry, we show that the removal of K+ binding site causes substantial changes in spin states of both the ferric and ferrous forms. The spectral changes are interpreted to be, most likely, due to the formation of a bis-histidine coordination structure in both the ferric and ferrous oxidation states at neutral pH 7.0. Stopped flow spectrophotometric techniques revealed that characteristics of Compound I were not observed in the K+ site double mutants in the presence of H2O2. Similarly electron donor oxidation rate was two orders less for the K+ site double mutants compared to the wild type. These data show that K+ functions in preserving the protein structure in the heme surroundings as well as the spin state of the heme iron, in favor of the enzymatically active form of LmP. [Copyright &y& Elsevier]

Abstract: The active site architecture of Leishmania major peroxidase (LmP) is very similar with both cytochrome c peroxidase and ascorbate peroxidase. We utilized point mutagenesis to investigate if the conserved proximal methionine residues (Met248 and Met249) in LmP help in controlling catalysis. Steady-state kinetics of methionine mutants shows that ferrocytochrome c oxidation is <2% of wild type levels without affecting the second order rate constant of first phase of Compound I formation, while the activity toward a small molecule substrate, guaiacol or iodide, increases. Our diode array stopped-flow spectral studies show that the porphyrin π-cation radical of Compound I in mutant LmP is more stable than wild type enzyme. These results suggest that the electronegative sulfur atoms of the proximal pocket are critical factors for controlling the location of a stable Compound I radical in heme peroxidases and are important in the oxidation of ferrocytochrome c. [Copyright &y& Elsevier]

Numerous reports have shown that mitochondrial dysfunctions play a major role in apoptosis of Leishmania parasites, but the endoplasmic reticulum (ER) stress-induced apoptosis in Leishmania remains largely unknown. In this study, we investigate ER stress-induced apoptotic pathways in Leishmania major using tunicamycin as an ER stress inducer. ER stress activates the expression of ER-localized chaperone protein BIP/GRP78 (binding protein/identical to the 78-kDa glucose-regulated protein) with concomitant generation of intracellular reactive oxygen species. Upon exposure to ER stress, the elevation of cytosolic Ca2+ level is observed due to release of Ca2+ from internal stores. Increase in cytosolic Ca2+ causes mitochondrial membrane potential depolarization and ATP loss as ablation of Ca2+ by blocking voltage-gated cation channels with verapamil preserves mitochondrial membrane potential and cellular ATP content. Furthermore, ER stress-induced reactive oxygen species (ROS)-dependent release of cytochrome c and endonuclease G from mitochondria to cytosol and subsequent translocation of endonuclease G to nucleus are observed. Inhibition of caspase-like proteases with the caspase inhibitor benzyloxycarbonyl-VAD-fluoromethyl ketone or metacaspase inhibitor antipain does not prevent nuclear DNA fragmentation and phosphatidylserine exposure. Conversely, significant protection in tunicamycin-induced DNA degradation and phosphatidylserme exposure was achieved by either pretreatment of antioxidants (N-acetyl-L-cysteine, GSH, and L-cysteine), chemical chaperone (4-phenylbutyric acid), or addition of Ca2+ chelator (1,2-bis(2-aminophenoxy)ethane-N,JV,N,N-tetraacetic acidacetoxymethyl ester). Taken together, these data strongly demonstrate that ER stress-induced apoptosis in L. major is dependent on ROS and Ca2+-induced mitochondrial toxicity but independent of caspase-like proteases. [ABSTRACT FROM AUTHOR]

Background: Peroxidase represents a heterogeneous group of distinct enzyme family that plays extremely diverse biological functions. Ascorbate peroxidase from Leishmania major (LmAPX) has been shown to be central to the redox defense system of Leishmania. To investigate further its exact physiological role in Leishmania, we attempted to create LmAPX -knockout mutants by gene replacement in L. major strains. Methodology/Principal Findings: The null mutant cell culture contains a higher percentage of metacyclic and apoptotic cells compared to both wild type and LmAPX overexpressing cells. Flowcytometric analysis reveals the presence of a higher concentration of intracellular H2O2, indicative of increased oxidative stress in parasites lacking LmAPX. IC50 value for exogenously added H2O2 shows that deletion of LmAPX in L. major renders the cell more susceptible to H2O2. Real time PCR studies demonstrate an elevated mRNA level of non-selenium glutathione peroxidase in LmAPX null mutant cell line, suggesting that these enzymes were induced to compensate the LmAPX enzyme. The null mutant cells exhibit hypervirulence after infection with macrophages as well as inoculation into BALB/c mice; in contrast, overexpressing cells show avirulence. Conclusions/Significance: Collectively, these data provide strong evidence that LmAPX is an important factor for controlling parasite differentiation and survival within macrophages. [ABSTRACT FROM AUTHOR]

Summary Cerebral cavernous malformations (CCMs) are neurovascular lesions caused by mutations in one of three genes (CCM1–3). Loss of CCM3 causes the poorest prognosis, and little is known about how it regulates vascular integrity. The C. elegans ccm-3 gene regulates the development of biological tubes that resemble mammalian vasculature, and in a genome-wide reverse genetic screen, we identified more than 500 possible CCM-3 pathway genes. With a phenolog-like approach, we generated a human CCM signaling network and identified 29 genes in common, of which 14 are required for excretory canal extension and membrane integrity, similar to ccm-3. Notably, depletion of the MO25 ortholog mop-25.2 causes severe defects in tube integrity by preventing CCM-3 localization to apical membranes. Furthermore, loss of MO25 phenocopies CCM3 ablation by causing stress fiber formation in endothelial cells. This work deepens our understanding of how CCM3 regulates vascular integrity and may help identify therapeutic targets for treating CCM3 patients. Graphical Abstract Highlights • Over 500 cerebral cavernous malformation (CCM) modifier genes discovered in C. elegans • Bioinformatics methods predict 29 conserved genes in core CCM-3/CCM3 network • Many genes exhibit ccm-3 phenotypes, including mop-25.2 • Loss of m op-25.2 homolog MO25 causes stress fiber formation in endothelial cells Lant et al. use C. elegans genetics to better understand the human disease cerebral cavernous malformation (CCM). Through a whole-genome screen, and bioinformatics, they uncover a set of conserved genes that exhibit ccm-3 phenotypes. These targets may be able to inform therapeutic studies. [ABSTRACT FROM AUTHOR]

Abstract: Architecture of hemoprotein is solely responsible for different nature of heme coordination. Here we report that substitution of the acidic surface residue Glu226 to Ala in ascorbate peroxidase from Leishmania major alters the 5 coordinate high spin (5cHS) to a 6 coordinate low spin (6cLS) form at pH 7.5. Using UV–visible spectrophotometry, we show that the sixth ligand of heme in Glu226Ala at pH 7.5 is hydroxo. When the pH is decreased to 5.5, a new species of Glu226Ala appeared that had a spectrum characteristic of a 6cHS derivative. Stopped flow spectrophotometric techniques revealed that characteristics of Compound I was not seen in the Glu226Ala in presence of H2O2. Similarly guaiacol, ascorbate and ferrocytochrome c oxidation rate was 103 orders less for the Glu226Ala mutants compared to the wild type. These data suggested that surface acidic residue Glu226 might play role in proper maintenance of active site conformation. [Copyright &y& Elsevier]

In this manuscript we demonstrate that a modification principally directed toward the improvement of the aqueous solubility (i.e., introduction a P3 pyridine N-oxide) of the previous lead compound afforded a new series of potent orally bioavailable P1 N-benzylamide thrombin inhibitors. An expedited investigation of the P1 SAR with respect to oral bioavailability, plasma half-life, and human liver microsome stability revealed 5 as the best candidate for advanced evaluation. [Copyright &y& Elsevier]