Developmental dynamics of mitochondrial fission and fusion proteins in functionally divergent skeletal muscles of goat.

During skeletal muscle development, the intricate mitochondrial network formation relies on continuous fission and fusion. This process in larger mammals differs from rodents, the most used animal models. However, the expression pattern of proteins regulating mitochondrial dynamics in developing skeletal muscle remains unexplored in larger mammals. Therefore, we characterized the cellular expression and tissue-level distribution of these proteins during development taking goat as a model. We have performed histological and immunohistochemical analyses to study metabolic features in various muscles. Neonatal muscles display uniform distribution of mitochondrial activity. In contrast, adult muscles exhibit clear distinctions based on their function, whether dedicated for posture maintenance or facilitating locomotion. Mitochondrial fission proteins like DRP-1, MFF, and fusion proteins like MFN-1 and 2 are abundantly expressed in neonatal muscles. Fission proteins exhibit drastic downregulation with limited peripheral expression, whereas fusion proteins continue to express in a fiber-specific manner during adulthood. Locomotory muscles exhibit different fibers based on mitochondrial activity and peripheralization with high SDH activity. The proximity ligation assay between MFN1 and MFN2 demonstrates that their interaction is restricted to subsarcolemmal mitochondria in adult fibers while distributed evenly in neonatal fibers. These differences between postural and locomotory muscles suggest their physiological and metabolic properties are different.

l-Asparaginase producing novel Streptomyces sp. HB2AG: optimization of process parameters and whole genome sequence analysis.

l-asparaginase (ASNase) is a key enzyme widely used as an anti-cancer drug and is also used in the pharmaceutical and food processing industries. This enzyme's applications are determined by its source and nature. The production of the enzyme through the fermentation process is also crucial for economic feasibility. Searching for a new potent microbial strain is necessary for increased ASNase synthesis. In this work, a potent strain was isolated from the sediment of Chilika Lake and selected for its high ASNase production potential. It was recognized following Bergey's manual of determinative and phylogenetic analysis was carried out by 16S rDNA sequencing. The isolated organism was Streptomyces sp. HB2AG. Additionally, a genome-wide analysis of HB2AG was performed. The result showed that the HB2AG genome possesses a chromosome with 6,099,956 bp and GC content of 74.0%. The whole genome analysis of the strain HB2AG revealed the presence of ASNase (ansA, ansB) and Asparagine synthase (asnB) in the HB2AG genome. Optimization of media composition is crucial for microbial growth and obtaining the desired end product. The current effort focuses on the Taguchi orthogonal design to determine optimum factor combinations that would allow the strain to produce maximum ASNase enzyme. Results showed that compared to unoptimized media, approximately 1.76-fold higher ASNase production was observed in Sea Water Luria Bertani (SWLB) media, pH-5, 0.5% (w/v) of lactose, 0.5% (w/v) of casein, 2.5% (w/v) NaCl, 1 mM Ca2+ and 0.1% Tween 80. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-023-03620-0.

High Tregs and systemic IL-10 expressions linked to the absence of sheath antibodies in lymphatic filariasis: implications on the persistence of residual infection.

The persistence of residual infection is one of the major factors in failure of the Global Programme to Eliminate Lymphatic Filariasis (GPELF). The present study aims to explore the status of sheath antibody and regulatory T cells (Tregs) known to play key roles in clearance of parasite and patent filarial infection, in individuals with residual infection after MDA. A total of 61 microfilaremic (Mf) individuals were followed up after at least 6 rounds of MDA. Infection status of subjects was assessed through the detection of Mf and circulating filarial antigen (CFA). Antibodies to Mf sheath were determined by immuno-peroxidase assay (IPA). The expression of Tregs was measured by a flow cytometer. IL-10 and IFN-γ were evaluated using the commercially available ELISA kit. The sheath antibody was present in subjects who have cleared both Mf and CFA and absent in individuals who were found to be Mf /CFA positive. Further individuals carrying infection have significantly high levels of Tregs and IL-10. A positive correlation was observed between Tregs, IL-10, and CFA in infected individuals. In contrast, a negative correlation was observed between IFN-γ and IL-10 in both infected and uninfected subjects. Our study reveals that the absence of a sheath antibody and a high level of Tregs and IL-10 are the hallmarks of the persistence of residual filarial infection.

Development of a colloidal gold strip-based immunochromatographic assay for rapid detection of Fusarium oxysporum in ginger.

BACKGROUND: Rhizome rot, caused primarily by Fusarium oxysporum, is one of the most destructive diseases leading to significant loss in ginger worldwide. The loss can be greatly reduced by proper disease management practices steered by accurate and early diagnosis of pathogens. Pathogen detection at an early stage of infection can also reduce the incidence of disease epidemics. Classical methods are often time consuming, relying on culturing the putative pathogens and the availability of expert taxonomic skills for accurate identification, which leads to the delayed application of control measures. The development of a simple, rapid, sensitive and cost-effective point-of-care diagnostic tool is thus one of the major research priorities for rhizome rot. RESULTS: The 65 kDa, immunoreactive protein band was selected as a diagnostic marker and was subjected to MS analysis followed by blastp. Based on blast result, a synthetic antigenic peptide was synthesized, and used to generate pAbs. The peptide-specific antibodies were used to develop a colloidal gold immunochromatographic assay (ICA). The sensitivity, specificity, and accuracy of ICA were 92.59%, 81.25%, and 90%, respectively. The ICA has a visual detection limit of 2.122 µg mL-1 for infected rhizome samples and 5.065 µg mL-1 for leaf samples with optimal detection time within 5 min. Moreover, the ICA also detected early stage infected samples, of which 71.42% (50/70) were true positives. CONCLUSION: Findings from this study indicated that the assay can be utilized as a tool for the investigation of rhizome rot infection in field samples. © 2019 Society of Chemical Industry.

Impact of novel N-aryl piperamide NO donors on NF-κB translocation in neuroinflammation: rational drug-designing synthesis and biological evaluation.

NO donor drugs showed a significant therapeutic effect in the treatment of many diseases, such as arteriopathies, various acute and chronic inflammatory conditions, and several degenerative diseases. NO-releasing anti-inflammatory drugs are the prototypes of a novel class of compounds, combining the pharmacological activities of anti-inflammatory and anti-nociceptive of drugs with those of NO, thus possessing potential therapeutic applications in a great variety of diseases. In this study, we designed and predicted biological activity by targeting cyclooxygenase type 2 (COX-2) and NF-κB subunits and pharmacological profiling along with toxicity predictions of various N-aryl piperamides linked via an ester bond to a spacer that is bound to a NO-releasing moiety (-ONO2). The result of absorption, distribution, metabolism and excretion and Docking studies indicated that among 51 designed molecules PA-3'K showed the best binding potential in both the substrate and inhibitory binding pocket of the COX-2 enzyme with affinity values of -9.33 and -5.12 for PDB ID 1CVU and 3LN1, respectively, thereby having the potential to be developed as a therapeutic agent. The results of cell viabilities indicated that PA-3'k possesses the best cell viability property with respect to its dose (17.33 ng/ml), with 67.76% and 67.93% viable cells for CHME3 and SVG cell lines, respectively.

Fungal disease detection in plants: Traditional assays, novel diagnostic techniques and biosensors.

Fungal diseases in commercially important plants results in a significant reduction in both quality and yield, often leading to the loss of an entire plant. In order to minimize the losses, it is essential to detect and identify the pathogens at an early stage. Early detection and accurate identification of pathogens can control the spread of infection. The present article provides a comprehensive overview of conventional methods, current trends and advances in fungal pathogen detection with an emphasis on biosensors. Traditional techniques are the "gold standard" in fungal detection which relies on symptoms, culture-based, morphological observation and biochemical identifications. In recent times, with the advancement of biotechnology, molecular and immunological approaches have revolutionized fungal disease detection. But the drawback lies in the fact that these methods require specific and expensive equipments. Thus, there is an urgent need for rapid, reliable, sensitive, cost effective and easy to use diagnostic methods for fungal pathogen detection. Biosensors would become a promising and attractive alternative, but they still have to be subjected to some modifications, improvements and proper validation for on-field use.

Maternal Filarial Infection Influences the Development of Regulatory T Cells in Children from Infancy to Early Childhood.

BACKGROUND: Children born from filarial infected mothers are comparatively more susceptible to filarial infection than the children born to uninfected mothers. But the mechanism of such increased susceptibility to infection in early childhood is not exactly known. Several studies have shown the association of active filarial infection with T cell hypo-responsiveness which is mediated by regulatory T cells (Tregs). Since the Tregs develop in the thymus from CD4+ CD25hi thymocytes at an early stage of the human fetus, it can be hypothesized that the maternal infection during pregnancy affects the development of Tregs in children at birth as well as early childhood. Hence the present study was designed to test the hypothesis by selecting a cohort of pregnant mothers and children born to them subsequently in a filarial endemic area of Odisha, India. METHODOLOGY AND PRINCIPAL FINDING: A total number of 49 pregnant mothers and children born to them subsequently have been followed up (mean duration 4.4 years) in an area where the microfilariae (Mf) rate has come down to <1% after institution of 10 rounds of annual mass drug administration (MDA). The infection status of mother, cord and children were assessed through detection of microfilariae (Mf) and circulating filarial antigen (CFA). Expression of Tregs cells were measured by flow cytometry. The levels of IL-10 were evaluated by using commercially available ELISA kit. A significantly high level of IL-10 and Tregs have been observed in children born to infected mother compared to children of uninfected mother at the time of birth as well as during early childhood. Moreover a positive correlation between Tregs and IL-10 has been observed among the children born to infected mother. SIGNIFICANCE: From these observations we predict that early priming of the fetal immune system by filarial antigens modulate the development of Tregs, which ultimately scale up the production of IL-10 in neonates and creates a milieu for high rate of acquisition of infection in children born to infected mothers. The mechanism of susceptibility and implication of the results in global elimination programme of filariasis has been discussed.

Emergence and spread of tetracycline resistant Vibrio cholerae O1 El Tor variant during 2010 cholera epidemic in the tribal areas of Odisha, India.

OBJECTIVES: The epidemics of cholera were reported in the Kashipur, K.singhpur, B cuttack blocks of Rayagada district and Mohana block of Gajapati district of Odisha during 2010. The present study was carried out to isolate the bacterial pathogen, its drug sensitivity pattern and to describe the spread of the disease in those areas. METHODS: A total of 68 rectal swabs collected from patients with severe diarrhea, admitted to different health centers and diarrhea affected villages were bacteriologically analyzed. Similarly 22 water samples collected from different villages from nala, chua, etc were tested for the presence of V cholerae. RESULTS: Out of 68 rectal swabs tested 35 (51.5%) were V cholerae O1 Ogawa and 14(20.6%) were E coli; which might be commensals. All water samples were negative for V cholerae. The V cholerae strains were sensitive to gentamicin, norfloxacin, ciprofloxacin, azithromycin and ofloxacin; but were resistant to ampicillin, tetracycline, nalidixic acid, furazolidone, streptomycin, erythromycin, co-trimoxazole, neomycin and chloramphenicol. All V cholerae strains were 100% resistant to tetracycline and they were El Tor variants harboring ctxB gene of classical strain. CONCLUSIONS: The present study indicated the emergence and spread of tetracycline resistant V cholerae O1 El Tor variant in the tribal areas which needs close monitoring.