Regulation of Neuronal Oxygen Responses in C. elegans Is Mediated through Interactions between Globin 5 and the H-NOX Domains of Soluble Guanylate Cyclases.

Soluble guanylate cyclases (sGCs) are gas-binding proteins that control diverse physiological processes such as vasodilation, platelet aggregation, and synaptic plasticity. In the nematode Caenorhabditis elegans, a complex of sGCs, GCY-35 and GCY-36, functions in oxygen (O2) sensing. Previous studies suggested that the neuroglobin GLB-5 genetically interacts with GCY-35, and that the inhibitory effect of GLB-5 on GCY-35 function is necessary for fast recovery from prolonged hypoxia. In this study, we identified mutations in gcy-35 and gcy-36 that impact fast recovery and other phenotypes associated with GLB-5, without undermining sGC activity. These mutations, heb1 and heb3, change conserved amino acid residues in the regulatory H-NOX domains of GCY-35 and GCY-36, respectively, and appear to suppress GLB-5 activity by different mechanisms. Moreover, we observed that short exposure to 35% O2 desensitized the neurons responsible for ambient O2 sensing and that this phenomenon does not occur in heb1 animals. These observations may implicate sGCs in neuronal desensitization mechanisms far beyond the specific case of O2 sensing in nematodes. The conservation of functionally important regions of sGCs is supported by examining site-directed mutants of GCY-35, which suggested that similar regions in the H-NOX domains of O2 and NO-sensing sGCs are important for heme/gas interactions. Overall, our studies provide novel insights into sGC activity and regulation, and implicate similar structural determinants in the control of both O2 and NO sensors. Significance statement: Soluble guanylate cyclases (sGCs) control essential and diverse physiological processes, including memory processing. We used Caenorhabditis elegans to explore how a neuroglobin inhibits a complex of oxygen-sensing sGCs, identifying sGC mutants that resist inhibition. Resistance appears to arise by two different mechanisms: increased basal sGC activity or disruption of an interaction with neuroglobin. Our findings demonstrate that the inhibition of sGCs by neuroglobin is essential for rapid adaptation to either low or high oxygen levels, and that similar structural regions are key for regulating both oxygen and nitric oxide sensors. Based on our structural and functional analyses, we present the hypothesis that neuroglobin-sGC interactions may be generally important for adaptation processes, including those in organisms with more complex neurological functions.

A simple, fast and cost-effective method of synthesis of cupric oxide nanoparticle with promising antibacterial potency: Unraveling the biological and chemical modes of action.

BACKGROUND: Gradual attainment of bacterial resistance to antibiotics led us to develop a robust method of synthesis of stable, colloidal cupric oxide nanoparticle of physiological pH with potential antibacterial action. METHODS: Cu(II) oxide NP was synthesized by reduction-oxidation of CuCl2, using polyvinyl alcohol as stabilizer. Characteristics and antibacterial activity of the particles were investigated by techniques like UV-Vis spectrophotometry, DLS, AFM, TEM, EDS, FTIR, AAS, agar plating, FACS, gel electrophoresis and XPS. RESULTS: The NPs were about 50 nm in size and cubic in shape with two surface plasmon peaks at 266 and 370 nm and had semi-conducting behavior with a band gap of 3.40 and 3.96 eV. About 80% of precursor CuCl2 was converted to NP. The minimum inhibitory and the minimum bactericidal concentrations of CuO-NP were respectively 120 and 160 µg/mL for Escherichia coli and 180 and 195 µg/mL for Staphylococcus aureus in Luria-Bertani medium. In growth media, the NPs got modified by media organics with displacement of the stabilizer PVA molecules. This modified NP (around 240 nm) killed cells by generating ROS, which finally caused membrane lipid per-oxidation and chromosomal DNA degradation in NP-treated cells. CONCLUSION: Reports indicate that we are among the few who had prepared CuO-NP in colloidal form. The antibacterial potency of our particle in growth media was much promising than other reports. Our findings demonstrated that 'particle-specific' effect, not 'ion-specific' one, was responsible for the NP action. GENERAL SIGNIFICANCE: The NP may be used as a sterilizing agent in various bioprocesses and as substituent of antibiotics, after thorough toxicological study.

Excitation dependent multicolor emission and photoconductivity of Mn, Cu doped In2S3 monodisperse quantum dots.

Indium sulphide (In2S3) quantum dots (QDs) of average size 6 ± 2 nm and hexagonal nanoplatelets of average size 37 ± 4 nm have been synthesized from indium myristate and indium diethyl dithiocarbamate precursors respectively. The absorbance and emission band was tuned with variation of nanocrytal size from very small in the strong confinement regime to very large in the weak confinement regime. The blue emission and its shifting with size has been explained with the donor-acceptor recombination process. The 3d element doping (Mn(2+) and Cu(2+)) is found to be effective for formation of new emission bands at higher wavelengths. The characteristic peaks of Mn(2+) and Cu(2+) and the modification of In(3+) peaks in the x-ray photoelectric spectrum (XPS) confirm the incorporation of Mn(2+) and Cu(2+) into the In2S3 matrix. The simulation of the electron paramagnetic resonance signal indicates the coexistence of isotropic and axial symmetry for In and S vacancies. Moreover, the majority of Mn(2+) ions and sulphur vacancies (VS ) reside on the surface of nanocrystals. The quantum confinement effect leads to an enhancement of band gap up to 3.65 eV in QDs. The formation of Mn 3d levels between conduction band edge and shallow donor states is evidenced from a systematic variation of emission spectra with the excitation wavelength. In2S3 QDs have been established as efficient sensitizers to Mn and Cu emission centers. Fast and slow components of photoluminescence (PL) decay dynamics in Mn and Cu doped QDs are interpreted in terms of surface and bulk recombination processes. Fast and stable photodetctors with high photocurrent gain are fabricated with Mn and Cu doped QDs and are found to be faster than pure In2S3. The fastest response time in Cu doped QDs is an indication of the most suitable system for photodetector devices.

Mechanism of antibacterial activity of copper nanoparticles.

In a previous communication, we reported a new method of synthesis of stable metallic copper nanoparticles (Cu-NPs), which had high potency for bacterial cell filamentation and cell killing. The present study deals with the mechanism of filament formation and antibacterial roles of Cu-NPs in E. coli cells. Our results demonstrate that NP-mediated dissipation of cell membrane potential was the probable reason for the formation of cell filaments. On the other hand, Cu-NPs were found to cause multiple toxic effects such as generation of reactive oxygen species, lipid peroxidation, protein oxidation and DNA degradation in E. coli cells. In vitro interaction between plasmid pUC19 DNA and Cu-NPs showed that the degradation of DNA was highly inhibited in the presence of the divalent metal ion chelator EDTA, which indicated a positive role of Cu(2+) ions in the degradation process. Moreover, the fast destabilization, i.e. the reduction in size, of NPs in the presence of EDTA led us to propose that the nascent Cu ions liberated from the NP surface were responsible for higher reactivity of the Cu-NPs than the equivalent amount of its precursor CuCl2; the nascent ions were generated from the oxidation of metallic NPs when they were in the vicinity of agents, namely cells, biomolecules or medium components, to be reduced simultaneously.

A simple robust method for synthesis of metallic copper nanoparticles of high antibacterial potency against E. coli.

A method for preparation of copper nanoparticles (Cu-NPs) was developed by simple reduction of CuCl2 in the presence of gelatin as a stabilizer and without applying stringent conditions like purging with nitrogen. The NPs were characterized by spectrophotometry, dynamic light scattering, x-ray diffraction, transmission electron microscopy, atomic force microscopy and x-ray photoelectron spectroscopy. The particles were about 50-60 nm in size and highly stable. The antibacterial activity of this Cu-NP on Gram-negative Escherichia coli was demonstrated by the methods of agar plating, flow cytometry and phase contrast microscopy. The minimum inhibitory concentration (3.0 µg ml(-1)), minimum bactericidal concentration (7.5 µg ml(-1)) and susceptibility constant (0.92) showed that this Cu-NP is highly effective against E. coli at a much lower concentration than that reported previously. Treatment with Cu-NPs made E. coli cells filamentous. The higher the concentration of Cu-NPs, the greater the population of filamentous cells; average filament size varied from 7 to 20 µm compared to the normal cell size of ∼2.5 µm. Both filamentation and killing of cells by Cu-NPs (7.5 µg ml(-1)) also occurred in an E. coli strain resistant to multiple antibiotics. Moreover, an antibacterial effect of Cu-NPs was also observed in Gram-positive Bacillus subtilis and Staphylococcus aureus, for which the values of minimum inhibitory concentration and minimum bactericidal concentration were close to that for E. coli.

A physiological exploration on operational stance and occupational musculoskeletal problem manifestations amongst construction labourers of West Bengal, India.

BACKGROUND: A huge number of labourers work in the construction industry in India both in organized and unorganized sectors. The construction labourers most often work for an extended period of time and they are compelled to uphold altered static and dynamic operational stance in awkward positions during the complete period of work which raises the demand on the musculoskeletal system and may lead to work related musculoskeletal disorders (WRMSDs). OBJECTIVE: This study is intended to explore the operational stance and occupation related musculoskeletal manifestations amongst the construction labourers. One hundred sixty four male labourers from different construction sites in West Bengal were randomly taken for this study. METHODS: A modified Nordic questionnaire on MSD and the 12 item General Health Questionnaire (GHQ12) were administered on the construction labourers. Rapid Entire Body Assessment (REBA) and Ovako Work Analysis System (OWAS) methods were applied to analyze the operational stance. Finally, discomfort levels of the specific operational stance were calculated by the use of risk level and BPD scale. RESULTS: The study revealed that most of the construction labourers habitually worked in awkward operational stance and were affected by altering musculoskeletal manifestations like pain in low back, neck, and wrist. We also found that there is a significant (p< 0.05) association between the intensity of pain feeling, age, year of working experience and risk level of the individual working postures of the labourers. CONCLUSION: Appropriate work-rest schedule, amendments of some working techniques and use of some ergonomically designed equipment may lessen the WRMSDs and improve the health eminence of construction labourers in unorganized sectors.

Multichain models of conserved lattice gas.

Conserved lattice-gas models in one dimension exhibit absorbing state phase transition (APT) with simple integer exponents ß=1=ν=η, whereas the same on a ladder belong to directed percolation (DP) universality. We conjecture that additional stochasticity in particle transfer is a relevant perturbation and its presence on a ladder forces the APT to be in the DP class. To substantiate this we introduce a class of restricted conserved lattice-gas models on a multichain system (M×L square lattice with periodic boundary condition in both directions), where particles which have exactly one vacant neighbor are active and they move deterministically to the neighboring vacant site. We show that for odd number of chains, in the thermodynamic limit L→∞, these models exhibit APT at ρ_{c}=1/2(1+1/M) with ß=1. On the other hand, for even-chain systems transition occurs at ρ_{c}=1/2 with ß=1,2 for M=2,4, respectively, and ß=3 for M≥6. We illustrate this unusual critical behavior analytically using a transfer-matrix method.

Synergism between soluble guanylate cyclase signaling and neuropeptides extends lifespan in the nematode Caenorhabditis elegans.

Oxygen (O2 ) homeostasis is important for all aerobic animals. However, the manner by which O2 sensing and homeostasis contribute to lifespan regulation is poorly understood. Here, we use the nematode Caenorhabditis elegans to address this question. We demonstrate that a loss-of-function mutation in the neuropeptide receptor gene npr-1 and a deletion mutation in the atypical soluble guanylate cyclase gcy-35 O2 sensor interact synergistically to extend worm lifespan. The function of npr-1 and gcy-35 in the O2 -sensing neurons AQR, PQR, and URX shortens the lifespan of the worm. By contrast, the activity of the atypical soluble guanylate cyclase O2 sensor gcy-33 in these neurons is crucial for lifespan extension. In addition to AQR, PQR, and URX, we show that the O2 -sensing neuron BAG and the interneuron RIA are also important for the lifespan lengthening. Neuropeptide processing by the proprotein convertase EGL-3 is essential for lifespan extension, suggesting that the synergistic effect of joint loss of function of gcy-35 and npr-1 is mediated through neuropeptide signal transduction. The extended lifespan is regulated by hypoxia and insulin signaling pathways, mediated by the transcription factors HIF-1 and DAF-16. Moreover, reactive oxygen species (ROS) appear to play an important function in lifespan lengthening. As HIF-1 and DAF-16 activities are modulated by ROS, we speculate that joint loss of function of gcy-35 and npr-1 extends lifespan through ROS signaling.

Mechanisms of defense against products of cysteine catabolism in the nematode Caenorhabditis elegans.

Cysteine catabolism presents cells with a double-edged sword. On the one hand, cysteine degradation provides cells with essential molecules such as taurine and sulfide. The formation of sulfide in cells is thought to regulate important and diverse physiological processes including blood circulation, synaptic activity and inflammation. On the other hand, the catabolism of cysteine by gut microbiota can release high levels of sulfide that may underlie the development or relapse of ulcerative colitis, an inflammatory bowel disease affecting millions of people worldwide. Here, we have used the nematode C. elegans to explore how cells tolerate high levels of sulfide produced by cysteine degradation in bacteria. We have identified mutations in genes coding for thioredoxin family proteins, mitochondrial proteins, and collagens that confer tolerance to sulfide toxicity. Exposure to sulfide induces the unfolded protein response in the endoplasmic reticulum and mitochondria. Moreover, our results suggest that sulfide toxicity is mediated by reactive oxygen species (ROS). Indeed, pre-treatment of worms with antioxidants increases their tolerance to sulfide toxicity. Intriguingly, sub-toxic levels of the superoxide generator paraquat can also increase the tolerance of worms to sulfide. Therefore, it appears that activation of ROS detoxification pathway prior to the exposure to sulfide, can increase the tolerance to sulfide toxicity. Our results suggest that these detoxification pathways are mediated by the hypoxia inducible factor HIF-1. Finally, we show that sulfide resistance varies among wild C. elegans and other nematode species, suggesting that tolerance to sulfide was naturally selected in certain habitats.

Multicritical absorbing phase transition in a class of exactly solvable models.

We study diffusion of hard-core particles on a one-dimensional periodic lattice subjected to a constraint that the separation between any two consecutive particles does not increase beyond a fixed value n+1; an initial separation larger than n+1 can however decrease. These models undergo an absorbing state phase transition when the conserved particle density of the system falls below a critical threshold ρ_{c}=1/(n+1). We find that the ϕ_{k}, the density of 0-clusters (0 representing vacancies) of size 0≤k

Prevalence of low back pain among handloom weavers in West Bengal, India.

BACKGROUND: Handloom is one of the oldest industries in India, particularly in West Bengal, where a considerable number of rural people are engaged in weaving. OBJECTIVES: The purpose of this study was to evaluate the prevalence of low back pain among the handloom weavers in India. METHODS: A modified Nordic Musculoskeletal Disorder Questionnaire and Oswestry Low Back Pain Disability Questionnaire along with a body part discomfort scale were administered to handloom weavers (n = 175). Working posture of the participants was assessed using the Ovako Working Posture Analysis System (OWAS). RESULTS: Sixty eight per cent of the participants reported suffering from low back pain, making it the most prevalent disorder in our sample. Analysis of the Oswestry Low Back Pain Disability Questionnaire data revealed that among those with low back pain (n = 119), 2% had severe disabilities, 46% had moderate disabilities, and 52% had minimal disabilities. Statistical analyses revealed a positive significant association between the intensity of pain in the lower back and an increased number of years of work experience (P<0·05). CONCLUSIONS: The study underlines the need for further research regarding the postural strain of weavers and also suggests the implementation of ergonomic design into weaver workstations to minimize the adverse effect of their current working postures. Improving upon the weaver's work-posture could improve their quality of life.

Calcium chloride made E. coli competent for uptake of extraneous DNA through overproduction of OmpC protein.

In the standard method of transformation of Escherichia coli with extraneous DNA, cells are made competent for DNA uptake by incubating in ice-cold 100 mM CaCl(2). Analysis of the whole protein profile of CaCl(2)-treated E. coli cells by the techniques of one- and two-dimensional gel electrophoresis, MALDI-MS and immunoprecipitation revealed overproduction of outer membrane proteins OmpC, OmpA and heat-shock protein GroEL. In parity, transformation efficiency of E. coli ompC mutant by plasmid pUC19 DNA was found to be about 40 % lower than that of the wild type strain. Moreover, in E. coli cells containing groEL-bearing plasmid, induction of GroEL caused simultaneous overproduction of OmpC. On the other hand, less OmpC was synthesized in E. coli groEL mutant compared to its wild type counterpart, by CaCl(2)-shock. From these results it can be suggested that in the process of CaCl(2)-mediated generation of competence, the heat-shock chaperone GroEL has specific role in DNA entry into the cell, possibly through the overproduced OmpC and OmpA porins.