Biological Control of <Emphasis Type="Italic">Fusarium</Emphasis> sp. NBRI-PMSF12 Pathogenic to Cultivated Betelvine by <Emphasis Type="Italic">Bacillus</Emphasis> sp. NBRI-W9, a Potential Biological Control Agent

Betelvine is prone to several fungal diseases including leaf spots, foot and root rot caused by Fusarium spp. due to humid conditions prevailing in fields. In the present study, a potent antagonistic bacterial endophyte and a virulent fungal pathogen were selected after rigorous screening of isolates from different betelvine varieties to provide an efficient biocontrol strategy in cultivation of betelvine. Wild varieties of crops are a rich source of untapped endophytes. Of the four betelvine varieties used for isolations and screening, the wild variety was richest in endophytic populations. Using 16S rRNA sequencing, the selected antagonist was identified as Bacillus sp. (NBRI-W9). The pathogen, virulent against cultivated varieties, was identified as Fusarium sp. (NBRI-PMSF12) using ITS 1 and 2 region sequencing. Under in vitro and field conditions, NBRI-W9 was able to induce early rooting, provide plant growth promotion, increase leaf size and yield (leaf number) and provide biocontrol against the Fusarium sp. infection. NBRI-W9 treatments showed bacterial colonization on the leaf surface preferably in the vicinity of pearl glands and the collenchyma region in scanning electron microscope (SEM) studies. NBRI-W9 was observed to directly enter the leaf by degrading cell walls and colonize the subcellular layers. SEM analysis showed direct confrontation of NBRI-W9 with Fusarium on the leaf surface and in the collenchyma region as one of the probable modes of biocontrol.     

Trace Mineral Micronutrients and Chronic Periodontitis—a Review

Trace mineral micronutrients are imperative for optimum host response. Populations worldwide are prone to their insufficiency owing to lifestyle changes or poor nutritional intake. Balanced levels of trace minerals like iron (Fe), zinc (Zn), selenium (Se) and copper (Cu) are essential to prevent progression of chronic conditions like periodontitis. Their excess as well as deficiency is detrimental to periodontal health. This is specifically true in relation to Fe. Furthermore, some trace elements, e.g. Se, Zn and Cu are integral components of antioxidant enzymes and prevent reactive oxygen species induced destruction of tissues. Their deficiency can worsen periodontitis associated with systemic conditions like diabetes mellitus. With this background, the present review first focusses on the role of four trace minerals, namely, Fe, Zn, Se and Cu in periodontal health followed by an appraisal of the data from case control studies related to their association with chronic periodontitis.     

Topological and epitope mapping of the cellular retinaldehyde-binding protein from retina.

Cellular retinaldehyde-binding protein (CRALBP) carries 11-cis-retinol or 11-cis-retinaldehyde as endogenous ligands and may function as a substrate carrier protein that modulates interaction of these retinoids with visual cycle enzymes. As a first approach to identifying functional domains and protein recognition sites in CRALBP, a low resolution topological and epitope map has been developed using monoclonal and polyclonal antibodies and limited proteolysis. Fifteen peptides of 8-31 residues spanning 99% of the 316-residue bovine CRALBP were synthesized and used to prepare 13 anti-peptide polyclonal antibodies. Using a competitive ELISA procedure, peptide epitopes were classified as either accessible or inaccessible in the native protein based on the extent of their recognition by these site-specific antibodies. Use of the synthetic peptides to map the epitopes of a polyclonal antibody to intact CRALBP confirmed that the amino terminus and carboxyl terminus are immunodominate regions and hence likely to be exposed, at least in part. Limited tryptic proteolysis of native CRALBP produced three major fragments which were shown by microsequence and Western analysis to be derived from sequential loss of short peptides from the amino terminus. None of these major fragments reacted with four monoclonal antibodies (mAbs) to intact CRALBP although each mAb immunoprecipitated native CRALBP. These results and the lack of mAb recognition of any of the synthetic peptides indicates that the amino terminus of the protein is exposed and contains part of an assembly epitope recognized by the mAbs. Overall this study indicates that residues 1-30, 100-124, and 257-285 contain highly exposed segments in the native protein and therefore constitute potential interaction domains for CRALBP and visual cycle enzymes. Residues 30-99 and 176-229 are inaccessible in the native structure and may be involved with retinoid binding. These results provide a basis for a systematic higher resolution mutagenesis study directed toward correlating CRALBP structural domains with function.     

Ocular distribution of 70-kDa heat-shock protein in rats with normal and dystrophic retinas

Summary Stress proteins are thought to play an important role in cellular development and in survival mechanisms. We compared the immunolocalization of the 70-kDa stress protein (SP70) in the ocular tissue of the normal Sprague-Dawley (SD) rat with that in the Royal College of Surgeons (RCS) rat with retinal dystrophy. SP70 was present in the maturing ocular tissues of both rat strains. However, once retinal degeneration began in the RCS rat, the retinal pigment epithelium and photoreceptor cells showed increased immunostaining for SP70 over that observed in age-matched SD rats. In late stages of retinal degeneration, immunostaining for SP70 was considerably reduced in the RCS retina, whereas normal distribution of immunostaining for SP70 in the SD retina was preserved, albeit decreased, through postnatal day 180. The optic nerve, ciliary body, and corneal epithelium were also influenced by the dystrophic disease condition, although the pattern of changes in SP70 immunostaining differed for each tissue. These results suggest that the genetic defect in the RCS rat produces a state of metabolic stress in all ocular tissues as the degeneration progresses, but that the subsequent rise in ocular SP70 is insufficient to prevent progression of the disease.     

Ecological studies on Ulva lactuca L. from Veraval,India

Ulva lactuca L., at Veraval (20° 54N and 70° 22E) on the western coast of India, grows in the intertidal belt from June to late February. During the summer months of March, April and May, the species dries up, leaving rhizoidal fragments for perennation. High values of density, phytomass, frequency, weight, and volume have been recorded during September and January. Accumulation of phytomass appears to be chiefly density dependent. High dissolved oxygen content and low temperatures of the surface sea water favoured enhanced growth and, consequently, more phytomass.     

Management of polycystic ovary syndrome among Indian women using myo‐inositol and D-chiro-inositol

Myo-Inositol and D-chiro-inositol (MI-DCI) are used in the treatment of polycystic Ovary syndrome (PCOS) due to their insulin-sensitizing actions. Therefore, it is of interest to evaluate the treatment patterns, clinical effectiveness and safety of MI-DCI combination in anagement of PCOS in Indian women. Data from 50 healthcare centers across India was collected between September 2019 and February 2020 and was used in the study. Women aged 12-45 years diagnosed with PCOS, who had received MI-DCI (550-150 mg) were included. The outcome parameters were change in weight, luteinizing hormone (LH)/follicle stimulating hormone (FSH) ratio, hirsutism, blood glucose and insulin levels, HOMA-IR, and lipid profile. A total of 283 women were included (mean age: 27.74 years; body mass index: 26.89 kg/m2); of which 197 (69.61%) reported reduction in weight after treatment with MI-DCI. The hirsutism scores considerably improved after treatment and the proportion of patients with, no hirsutism increased from 31.07% to 50.51% and moderate hirsutism reduced from 32.52% to 6.12% while, there were no patients with severe hirsutism after the treatment. There was a significant reduction in LH:FSH ratio (mean difference: 0.25 mg/dL; p=0.021), free testosterone (mean difference: 1.49; p<0.001) and dehydroepiandrosterone (mean difference: 21.49; P<0.001) levels after regular use of MI-DCI tablets. Treatment with MI-DCI resulted in significant improvement in insulin levels, HOMA-IR score, Fasting plasma glucose (FPG), post-prandial plasma glucose (PPG) and lipid profile. The therapy restored menstruation and spontaneous ovulation and significantly attenuated the LH/FSH ratio. Thus, MI-DCI (550-150 mg) has shown multidimensional benefits in improving the hormonal, glycemic, and lipid profile of women with PCOS with considerable efficacy and tolerability.     

An innovative and cost-effective way to estimate alkaline phosphatase activity in in vitro cellular model systems

Alkaline phosphatase is an enzyme that converts para-nitrophenyl phosphate to para-nitrophenol (yellow coloured) in 2-amino, 2-methyl, 1-propanol buffer at pH 10.5. However, when this protocol is applied to the in vitro cellular model systems to estimate alkaline phosphatase activity, it tends to generate clumps of genomic DNA, leading to inaccurate pipetting for protein estimation. The aim of the study was to introduce minor modifications in the existing protocol to make it simple, cost-effective, with minimal labor-intensive procedures while estimating alkaline phosphatase activity in cellular model systems. The genomic DNA clumps were dissolved by depurination (adding 0.2 N HCl) and fragmentation (adding 0.2 N NaOH) during enzyme estimation. Moreover, these minor modifications have been standardized and optimized extensively by using serum samples (rich source of alkaline phosphatase), hFOB/ER9 (human Fetal osteoblastic cell) and HepG2 cells. Our results suggest that the modification incorporated in previously published method was robust enough to estimate ALP activity and protein concentration accurately. There was no significant variation in ALP activity estimated after modification (P > 0.05). This innovative approach could be beneficial for a researcher by providing an easy, cost effective and less labor-intensive solution for estimation of enzymatic activity in cellular model systems.