Repolarization of the action potential enabled by Na<Superscript>+</Superscript> channel deactivation in PSpice simulation of cardiac muscle propagation

Background  

In previous studies on propagation of simulated action potentials (APs) in cardiac muscle using PSpice modeling, we reported that a second black-box (BB) could not be inserted into the K⁺ leg of the basic membrane unit because that caused the PSpice program to become very unstable. Therefore, only the rising phase of the APs could be simulated. This restriction was acceptable since only the mechanism of transmission of excitation from one cell to the next was being investigated.     

Feasibility studies on the treatment of dairy wastewaters with upflow anaerobic sludge blanket reactors

The feasibility of using upflow anaerobic sludge blanket (UASB) reactors for the treatment of dairy wastewaters was explored. Two types of UASBs were used--one operating on anaerobic sludge granules developed by us from digested cowdung slurry (DCDS) and the other on the granules obtained from the reactors of M/s EID Parry treating sugar industry wastewaters. The reactors were operated at HRT of 3 and 12 h and on COD loading rates ranging from 2.4 kg per m3 of digester volume, per day to 13.5 kg m(-3) d(-1). At the 3 h HRT, the maximum COD reduction in the DCDS-seeded and the industrial sludge-seeded reactors was 95.6% and 96.3%, respectively, better than at 12 h HRT (90% and 92%, respectively). In both the reactors, the maximum, the second best, and the third best COD reduction occurred at the loading rates of 10.8, 8.6 and 7.2 kg m3 d(-1), respectively. At loading rates higher than 10.8 kg, the reactor performance dropped precipitously. Whereas in the first few months the reactors operating on sludge from EID Parry achieved better biodegradation of the waste, compared to the reactors operated on DCDS, the performance of the latter gradually improved and matched with the performance of the former.     

Overexpression, purification, and pharmacological activity of a biosynthetically derived conopeptide

A high yielding fusion protein system based on the protein cytochrome b(5) has been used for the production of novel 13-residue acyclic conopeptide. This peptide, Mo1659, can be liberated from the carrier protein using CNBr cleavage and subsequent purification using RP-HPLC methods. The yield of isotopically enriched peptides is high, ranging from 3 to 4mg of purified peptide from a 500ml culture, indicating that this system can be widely used for peptide production. Biosynthetic Mo1659 is active on non-inactivating K(+) channel much like the natural Mo1659, despite the absence of C-terminal amidation. Heteronuclear NMR studies show that the peptide exists in a conformational equilibrium involving proline-10. To our knowledge this is the first report of the production of an isotopically (15)N/(13)C-enriched conopeptide.     

A PCR-based approach to sequence the Candida tropicalis HSP90 gene

The heat shock protein 90 (hsp90) gene sequence is known to be highly conserved across the species barrier. A PCR-based method was thus utilised in an attempt to sequence the Candida tropicalis hsp90 gene. Primers for PCR were designed from conserved regions of the gene, which were identified by comparing the Saccharomyces cerevisiae and Candida albicans hsp90 gene sequences. Different sets of primers were designed to amplify and obtain overlapping DNA sequences of the C tropicalis gene. PCR was carried out on genomic DNA of Candidca tropicalis and the PCR products were cloned into suitable vector molecules for sequencing. In this way, a 2,070-basepair sequence of the C. tropicalis hsp90 gene was obtained. The PCR-based approach proved to be an easier method of obtain the sequence of a highly conserved gene, as compared to more conventional methods.     

Peripheral plasma progesterone: a marker for determining cyclicity in yaks (Poephagus grunniens L.)

An attempt was made to determine cyclicity in yaks using plasma progesterone during the breeding and non-breeding seasons. Fifteen non-lactating yaks were used in this experiment. During the breeding season (July to November), blood samples were collected from 8 yaks at least twice weekly until estrus was observed and then at 2 days interval for 30 days. During the non-breeding season (February to March), blood samples were collected from the same number of yaks at 2-day interval for 30 days. Progesterone was determined in plasma samples by radioimmunoassay. During the breeding season, plasma progesterone at estrus was basal (< or = 0.2 ng/ml). Concentrations increased thereafter with a sharp increase during the late luteal phase, typically reaching peak levels around day 15. Concentrations then declined rapidly over the following 4 days, reaching basal levels at estrus. A high proportion (66.7%) of potential estrous periods (based on progesterone concentrations) went undetected, indicating that silent or weak estrus was a prominent problem in yak cows. During the non-breeding season, three animals were found to be cycling as determined by the patterns of plasma progesterone. Yet, behavioral symptoms of estrus were not observed in any of these yak cows. We conclude that peripheral plasma progesterone concentrations can be used to monitor cyclicity in yak cows effectively.     

Lipopolysaccharide-induced alterations in oxygen consumption and radical generation in endothelial cells

Oxygen consumption rate (OCR) and generation of superoxide and nitric oxide (NO) in mouse aortic endothelial cells (MAECs) treated with lipopolysaccharide (LPS) were studied. The OCR was determined in cell suspensions at 37 °C by electron paramagnetic resonance (EPR) spectroscopy. LPS significantly altered the OCR in a dose and time-dependent fashion. The OCR was significantly elevated immediately following the treatment of MAECs with LPS (5 and 10 μg/ml) and NADPH (100 μM) whereas the same was depressed 1 h after exposure to similar conditions of incubation. Under similar experimental conditions, superoxide generation was also determined by EPR spectroscopy and cytochrome c reduction assays. A marginal increase in the superoxide production was observed when the cells were treated with LPS and NADPH alone whereas the same was further enhanced significantly when the cells were treated with LPS and NADPH together. The increase in oxygen consumption and superoxide production caused by LPS was inhibited by diphenyleneiodonium (DPI), suggesting the involvement of NAD(P)H oxidase. A significant increase in the NO production by MAECs was noticed 1 h after treatment with LPS and was inhibited by L-NAME, further suggesting the involvement of nitric oxide synthase (NOS). Thus, on a temporal scale, LPS-induced alterations in oxygen consumption by MAECs may be under the control of dual regulation by NAD(P)H oxidase and NOS. (Mol Cell Biochem 278: 119–127, 2005)     

A database for Plasmodium falciparum protein models

There is an urgent need for developing alternate strategies to combat Malaria caused by Plasmodium falciparum (P. falciparum) because of growing drug resistance and increased incidents of infection in humans. 3D models of P. falciparum annotated proteins using molecular modeling techniques will enhance our understanding about the mechanism of host parasite interactions for the identification of drug targets and malarial vaccine design. Potential structural templates for P. falciparum annotated proteins were selected from PDB (protein databank) using BLASTP (basic local alignment search tool for proteins). This exercise identified 476 Plasmodium proteins with one or more known structural templates (>or= 40 % identity) for further modeling. The pair-wise sequence alignments generated for protein modeling were manually checked for error. The models were then constructed using MODELLER (a comparative protein modelling program for modelling protein structures) followed by energy minimization in AMBER force field and checked for error using PROCHECK. AVAILABILITY: http://bioinfo.icgeb.res.in/codes/model.html.     

Advanced glycation end products and RAGE: a common thread in aging, diabetes, neurodegeneration, and inflammation

The products of nonenzymatic glycation and oxidation of proteins and lipids, the advanced glycation end products (AGEs), accumulate in a wide variety of environments. AGEs may be generated rapidly or over long times stimulated by a range of distinct triggering mechanisms, thereby accounting for their roles in multiple settings and disease states. A critical property of AGEs is their ability to activate receptor for advanced glycation end products (RAGE), a signal transduction receptor of the immunoglobulin superfamily. It is our hypothesis that due to such interaction, AGEs impart a potent impact in tissues, stimulating processes linked to inflammation and its consequences. We hypothesize that AGEs cause perturbation in a diverse group of diseases, such as diabetes, inflammation, neurodegeneration, and aging. Thus, we propose that targeting this pathway may represent a logical step in the prevention/treatment of the sequelae of these disorders.     

Composting-vermicomposting of leaf litter ensuing from the trees of mango (Mangifera indica)

Litter of the mango (Mangifera indica) tree leaves was composted and then converted into vermicast by the action of the earthworm Eudrilus eugeniae Kinberg. After over nine months of continuous operation the vermireactors with 62.5 animals l(-1) generated approximately 13.6g vermicast per litre of reactor volume (l) per day (d) whereas the reactors with 75 animals l(-1) produced approximately 14.9 g vermicast l(-1) d(-1). This difference in performance of the reactors operating in duplicate at the two different earthworm densities was statistically significant (> or = 90% confidence level) for most of the nine-month span. The animals grew well in all reactors, increasing their zoomass by approximately 103% and producing approximately 157 offspring. Not a single of the 1100 animals died during the first four months. In the subsequent five months a total of 122 worms died, representing a loss of approximately 2% per month. We attribute this to the normal process of ageing. The ability of the earthworms to survive, grow and breed in the vermireactors fed with composted mango tree leaves, and a rising trend in vermicast output inspite of the death of a few worms after four months of reactor operation, indicate the sustainability of this type of vermireactors. The studies also indicate that even better vermireactor efficiency may be possible by modifying the reactor geometry. Studies on changes in C:N ratio during composting and vermicomposting revealed that whereas composting helped in lowering the ratio due to loss of carbon in bacterial metabolism, vermicomposting had no such effect on the ratio.     

Gap-junction channels inhibit transverse propagation in cardiac muscle

The effect of adding many gap-junctions (g-j) channels between contiguous cells in a linear chain on transverse propagation between parallel chains was examined in a 5 × 5 model (5 parallel chains of 5 cells each) for cardiac muscle. The action potential upstrokes were simulated using the PSpice program for circuit analysis. Either a single cell was stimulated (cell A1) or the entire chain was stimulated simultaneously (A-chain). Transverse velocity was calculated from the total propagation time (TPT) from when the first AP crossed a V_m of -20 mV and the last AP crossed -20 mV. The number of g-j channels per junction was varied from zero to 100, 1,000 and 10,000 (R_gj of ∞, 100 MΩ, 10 MΩ, 1.0 MΩ, respectively). The longitudinal resistance of the interstitial fluid (ISF) space between the parallel chains (R_ol2) was varied between 200 KΩ (standard value) and 1.0, 5.0, and 10 MΩ. The higher the R_ol2 value, the tighter the packing of the chains. It was found that adding many g-j channels inhibited transverse propagation by blocking activation of all 5 chains, unless R_ol2 was greatly increased above the standard value of 200 KΩ. This was true for either method of stimulation. This was explained by, when there is strong longitudinal coupling between all 5 cells of a chain awaiting excitation, there must be more transfer energy (i.e., more current) to simultaneously excite all 5 cells of a chain.