Low frequency and low intensity pulsed electromagnetic field exerts its antiinflammatory effect through restoration of plasma membrane calcium ATPase activity

Rheumatoid arthritis (RA) is a chronic inflammatory disorder affecting 1% of the population worldwide. Pulsed electromagnetic field (PEMF) has a number of well-documented physiological effects on cells and tissues including antiinflammatory effect. This study aims to explore the antiinflammatory effect of PEMF and its possible mechanism of action in amelioration of adjuvant induced arthritis (AIA). Arthritis was induced by a single intradermal injection of heat killed Mycobacterium tuberculosis at a concentration of 500 μg in 0.1 ml of paraffin oil into the right hind paw of rats. The arthritic animals showed a biphasic response regarding changes in the paw edema volume. During the chronic phase of the disease, arthritic animals showed an elevated level of lipid peroxides and depletion of antioxidant enzymes with significant radiological and histological changes. Besides, plasma membrane Ca²⁺ ATPase (PMCA) activity was inhibited while intracellular Ca²⁺ level as well as prostaglandin E₂ levels was noticed to be elevated in blood lymphocytes of arthritic rats. Exposure of arthritic rats to PEMF at 5 Hz × 4 μT × 90 min, produced significant antiexudative effect resulting in the restoration of the altered parameters. The antiinflammatory effect could be partially mediated through the stabilizing action of PEMF on membranes as reflected by the restoration of PMCA and intracellular Ca²⁺ levels in blood lymphocytes subsequently inhibiting PGE₂ biosynthesis. The results of this study indicated that PEMF could be developed as a potential therapy for RA in human beings.     

Evaluation of neuroprotective and anti-fatigue effects of sildenafil

Sildenafil, a phosphodiesterase-5 inhibitor is widely used for the treatment of erectile dysfunction. Recently, the FDA approved the use of sildenafil in the therapeutic treatment of pulmonary arterial hypertension. Sildenafil crosses the blood-brain barrier and has been shown to enhance memory. Tremor, rigidity and akinesia are the most common symptoms seen in Parkinson's disease. Fatigue and sexual dysfunction are the other prominent features seen in Parkinson's disease. Interestingly, sildenafil is used therapeutically to treat sexual dysfunction in Parkinson's disease patients. Currently research on Parkinson's disease focuses on developing novel drug therapies for retarding the nigral dopaminergic neurodegeneration. Hence, we investigated the anti-fatigue and neuroprotective effects of sildenafil. In this study, the effect of sildenafil on fatigue was evaluated using forced swim test in mice. Sildenafil had no effect on fatigue as seen by the swim time. With regard to neuroprotective effects, we investigated the effects of sildenafil using two animal models of Parkinson's disease. In this study, 6-hydroxydopamine-lesioned (unilateral) rats and MPTP-treated mice were used as the animal models of Parkinson's disease. 6-Hydroxydopamine-lesioned rats were used to determine the effect of sildenafil on rotational behavior. Ipsilateral or contralateral rotational behavior can indicate the amphetamine-like activity or apomorphine-like activity of sildenafil. Sildenafil did not induce contralateral or ipsilateral rotations in 6-hydroxydopamine-lesioned rats. Sildenafil did not protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced dopamine depletion in the striatum.     

Antifungal susceptibility profiles of <Emphasis Type="Italic">Coccidioides immitis</Emphasis> and <Emphasis Type="Italic">Coccidioides posadasii</Emphasis> from endemic and non-endemic areas

Coccidioidomycosis is a systemic fungal infection endemic in Southwestern United States, Mexico, Central and South America. The causal agents are Coccidioides immitis and C. posadasii. A large number of cases of coccidioidomycosis in New York State residents were identified. We compared susceptibility profiles of these isolates and of C. immitis isolates from California using mycelial phase inoculum and CLSI (NCCLS) M38–A broth microdilution protocol. Minimum fungicidal concentrations (MFC) were also determined. Results indicated that geometric mean MICs of amphotericin B (AMB, 0.06 μg/ml), fluconazole (FLC, 8.0 μg/ml), itraconazole (ITC, 0.07 μg/ml), ketoconazole (KTC, 0.04 μg/ml), voriconazole (VRC, 0.04 μg/ml), posaconazole (PSC, 0.17 μg/ml) and caspofungin (CSP, 0.15 μg/ml) were in susceptible range as per breakpoints published for pathogenic Candida species. However, geometric MFC for FLC was relatively higher (52.4 μg/ml). Also, no significant difference in MIC and MFC values was evident for C. immitis and C. posadasii isolates. In conclusion, current methods for antifungal susceptibility testing yield reproducible profiles for Coccidioides species, which appear to be highly susceptible to most antifungal agents.     

Comparison of mealybug (Planococcus lilacinus) and fruit fly genomes: isolation and analysis of conserved sequences and their utility in studying synteny in the mealybug

By using ligation-mediated PCR products from mealybug DNA as tester and biotinylated fly DNA as driver, we recovered a fraction of the tester that remains hybridized to driver following high-stringency washing conditions. This fraction is expected to contain mealybug sequences conserved in the fly (MCF). Reciprocal experiments enabled the isolation of fly sequences conserved in the mealybug (FCM). Coding sequences among MCF show amino acid identities >40% with fly proteins, allowing a reliable identification of orthologs. Three sequences from the fly cytogenetic positions 98-99 were hybridized onto mealybug chromosomes and the results identified differences in synteny between the two species. Taken together, our results present a method for direct isolation of sequences conserved between an 'orphan' (mealybug) genome and a 'reference' (fly) genome and showed that these sequences can be used to study chromosome synteny in the mealybug.     

An empirical approach for deriving specific inland water quality parameters from high spatio-spectral resolution image

Inland lake of Vembanad has benefited from continuous monitoring to evaluate water quality which has declined due to increased anthropogenic activities and climate change. Remote sensing techniques can be used to estimate and monitor inland water quality both spatially and temporally. An empirical model is presented in Vemaband lake that retrieves the specific water quality parameters through correlations between various spectral wavelengths of Sentinel-2MSI (S2MSI) with field-measured water quality parameters. This approach includes the combinations of various bands, band ratios, and band arithmetic computation of satellite sensors of spectral datasets. The specific inland water quality parameters such as chlorophyll-a (chl-a), total suspended solids (TSS), turbidity, and secchi disc depth (SDD) were retrieved from the developed water quality model through Sentinel-2A remote sensing reflectance. The result illustrates that Specific Inland Water Quality Parameters (SIWQP) strongly correlated with S2MSI reflection spectral wavelengths. The SIWQP models are constructed for TSS (R²?=?0.8008), Chl-a (R²?=?0.8055), Turbidity (R²?=?0.6329) and SDD (R²?=?0.7174).The spatial distribution of SIWQPs in Vembanad lake for March 2018 is mapped and shows the lake's water quality distribution. The research from Sentinel-2, MSI has potential and is appropriate in high spectral and spatial characteristics for retrieving and continuous monitoring of water quality parameters in the regional scale of inland water bodies.

     

Multiplex gene removal by two-step polymerase chain reactions

Precise DNA manipulation is critical for molecular biotechnology. Restriction enzyme-based approaches are limited by their requirement of specific enzyme sites. Restriction-free cloning has greatly improved the flexibility and speed of precise DNA assembly. Most of these approaches focus on DNA assembly rather than gene removal. Here we present a polymerase chain reaction (PCR)-based cloning method that allows removal of multiple gene segments from plasmids without using restriction enzymes and thermostable ligase. We demonstrate simultaneous removal of three gene segments from a plasmid. This approach could be beneficial to DNA library construction, genetic and protein engineering, and synthetic biology.     

Design and synthesis of 3-(azol-1-yl)phenylpropanes as microbicidal spermicides for prophylactic contraception

We designed a series of 25 3-(azol-1-yl)phenylpropanes which yielded 10 compounds (3, 4, 7, 8, 13, 14, 19, 21, 23, 26) that irreversibly immobilized 100% human sperm at 1% (w/v) concentration in 60 s; 12 compounds (8, 9, 15, 16, 19-21, 23-25, 27, 28) that showed potent microbicidal activity at 12.5-50 μg/mL against Trichomonas vaginalis; and 17 compounds (3-11, 13, 15, 19, 21, 23, 26, 28, 30) that exhibited potent anticandida activity with minimum inhibitory concentration (MIC) of 12.5-50 μg/mL. Almost all the compounds exhibited high level of safety towards normal vaginal flora (Lactobacillus) and human cervical (HeLa) cells in comparison to the marketed spermicide nonoxynol-9 (N-9). All the biological activities were evaluated in vitro. Two compounds (4, 8) with good safety profile exhibited multiple (spermicidal, antitrichomonas and anticandida) activities, warranting further lead optimization for furnishing a prophylactic vaginal contraceptive.     

Benzothiophene carboxamide derivatives as inhibitors of Plasmodium falciparum enoyl-ACP reductase

Benzothiophene derivatives like benzothiophene sulphonamides, biphenyls, or carboxyls have been synthesized and have found wide pharmacological usage. Here we report, bromo-benzothiophene carboxamide derivatives as potent, slow tight binding inhibitors of Plasmodium enoyl-acyl carrier protein (ACP) reductase (PfENR). 3-Bromo-N-(4-fluorobenzyl)-benzo[b]thiophene-2-carboxamide (compound 6) is the most potent inhibitor with an IC50 of 115 nM for purified PfENR. The inhibition constant (Ki) of compound 6 was 18 nM with respect to the cofactor and 91 nM with respect to crotonoyl-CoA. These inhibitors showed competitive kinetics with cofactor and uncompetitive kinetics with the substrate. Thus, these compounds hold promise for the development of potent antimalarials.