Reduced Pancreatic Exocrine Function and Organellar Disarray in a Canine Model of Acute Pancreatitis

The aim of the present study was to investigate the pancreatic exocrine function in a canine model and to analyze the changes in organelles of pancreatic acinar cells during the early stage of acute pancreatitis (AP). AP was induced by retrograde injection of 5% sodium taurocholate (0.5 ml/kg) into the main pancreatic duct of dogs. The induction of AP resulted in serum hyperamylasemia and a marked reduction of amylase activity in the pancreatic fluid (PF). The pancreatic exocrine function was markedly decreased in subjects with AP compared with the control group. After the induction of AP, histological examination showed acinar cell edema, cytoplasmic vacuolization, fibroblasts infiltration, and inflammatory cell infiltration in the interstitium. Electron micrographs after the induction of AP revealed that most of the rough endoplasmic reticulum (RER) were dilated and that some of the ribosomes were no longer located on the RER. The mitochondria were swollen, with shortened and broken cristae. The present study demonstrated, in a canine model, a reduced volume of PF secretion with decreased enzyme secretion during the early stage of AP. Injury of mitochondria and dilatation and degranulation of RER may be responsible for the reduced exocrine function in AP. Furthermore, the present model and results may be useful for researching novel therapeutic measures in AP.     

Murine GFP-Mx1 forms nuclear condensates and associates with cytoplasmic intermediate filaments: Novel antiviral activity against VSV

Type I and III interferons induce expression of the “myxovirus resistance proteins” MxA in human cells and its ortholog Mx1 in murine cells. Human MxA forms cytoplasmic structures, whereas murine Mx1 forms nuclear bodies. Whereas both HuMxA and MuMx1 are antiviral toward influenza A virus (FLUAV) (an orthomyxovirus), only HuMxA is considered antiviral toward vesicular stomatitis virus (VSV) (a rhabdovirus). We previously reported that the cytoplasmic human GFP-MxA structures were phase-separated membraneless organelles (“biomolecular condensates”). In the present study, we investigated whether nuclear murine Mx1 structures might also represent phase-separated biomolecular condensates. The transient expression of murine GFP-Mx1 in human Huh7 hepatoma, human Mich-2H6 melanoma, and murine NIH 3T3 cells led to the appearance of Mx1 nuclear bodies. These GFP-MuMx1 nuclear bodies were rapidly disassembled by exposing cells to 1,6-hexanediol (5%, w/v), or to hypotonic buffer (40–50 mosm), consistent with properties of membraneless phase-separated condensates. Fluorescence recovery after photobleaching (FRAP) assays revealed that the GFP-MuMx1 nuclear bodies upon photobleaching showed a slow partial recovery (mobile fraction: ∼18%) suggestive of a gel-like consistency. Surprisingly, expression of GFP-MuMx1 in Huh7 cells also led to the appearance of GFP-MuMx1 in 20–30% of transfected cells in a novel cytoplasmic giantin-based intermediate filament meshwork and in cytoplasmic bodies. Remarkably, Huh7 cells with cytoplasmic murine GFP-MuMx1 filaments, but not those with only nuclear bodies, showed antiviral activity toward VSV. Thus, GFP-MuMx1 nuclear bodies comprised phase-separated condensates. Unexpectedly, GFP-MuMx1 in Huh7 cells also associated with cytoplasmic giantin-based intermediate filaments, and such cells showed antiviral activity toward VSV.     

MimoSA: a system for minimotif annotation

Background  

Minimotifs are short peptide sequences within one protein, which are recognized by other proteins or molecules. While there are now several minimotif databases, they are incomplete. There are reports of many minimotifs in the primary literature, which have yet to be annotated, while entirely novel minimotifs continue to be published on a weekly basis. Our recently proposed function and sequence syntax for minimotifs enables us to build a general tool that will facilitate structured annotation and management of minimotif data from the biomedical literature.     

Compatibility testing and enhancing the pulp bleaching process by hydrolases of the newly isolated thermophilic Isoptericola variabilis strain UD-6

Abstract

In the present study, Isoptericola variabilis strain UD-6 isolated from alkaline hot spring of Unapdev, Maharashtra, India was assessed for its biobleaching activity by hydrolytic enzymes on rice straw pulp. Results of primary and secondary screening manifested that it was a multi-enzyme producer, competent to produce amylase, cellulase, mannanase, pectinase, and xylanase at 9.73, 4.11, 6.26, 8.42, and 6.61?IU?ml^?1 in fermentation conditions, respectively. Maximum activity of all enzymes was gained at thermal temperature (50–55?°C), alkaline condition (pH 8–9), under 5?mM KCl and 5?mM NaCl salt concentration. In compatibility testing, activities of all enzymes were spectacularly reduced when they utilized with chemicals of pulp bleaching. Results of rice straw pulp bleaching was effectual when pulp was initially bleached with mannanase, pectinase, and xylanase enzymes (Es) for 90?min and then with diluted chemicals (DC) for further 90?min instead of their separate use. Treatment of rice straw pulp with Es?+?DC, enhanced the release of reducing sugars, hydrophobic compounds, and phenolic compounds, whereas Kappa number was reduced. Overall, the results of the present study indicated that pre-bleaching of pulp with hydrolytic enzymes obtained from I. variabilis strain UD-6 helps to minimize chemicals used in the bleaching process and make it more sustainable for pulp and paper industries as well as for the environment.     

Carbon distribution to toxic effect in toxin proteins

The role of hydrophobic force in biological function through the formation of several local macro-molecular structures is evident. Carbon is the element that contributes to biological function in living systems. We show that carbon distribution is related to protein activity using an example. The carbon distribution profile is foreseen to help undestand unfolded and misfolded regions of protein structures. The carbon distribution profile in a toxin protein that is found associated with the toxic shock syndrome is described in this study. The carbon profile provides insight to the association of specific residues responsible for toxicity.     

Development of a photosensitive, high-throughput chip-based superoxide dismutase (SOD) assay to explore the radioprotective activity of herbal plants

Appropriate pharmacological interventions and modalities are needed to protect humans against the deleterious effects of ionizing radiation. We disclose a rapid chip-based approach to elucidate the radioprotective/antioxidant potential of herbal plants using a photodiode array (PDA) microchip system. Red light absorption property of nitroblue tetrazolium (NBT) formazan was applied to chip-based superoxide dismutase (SOD) activity measurements of six herbal plant extracts in a high-throughput manner. SOD activities obtained via gel-based assays were in line with the data obtained through the chip-based assay and hence validated our approach. Compared to amifostine, all the tested herbal plant extracts, except apricot kernel, demonstrated greater radioprotective properties. Among the tested herbal extracts, pueraria root showed the highest antioxidant/radioprotective activity and can be considered a preferred radioprotector candidate. Low standard deviations and high statistical confidence obtained during the assay prove the sensitivity and consistency of this approach. The developed approach has several advantages (simplicity, rapidness and portability) over existing methods and can be applied to high-throughput screening of the radioprotective properties of various unexplored plants species.     

Temperature and Photoperiod Influence Postharvest Needle Abscission of Selected Balsam Fir (Abies balsamea L. (Mill.)) Genotypes by Modulating ABA Levels

Seasonal changes in the concentration of abscisic acid (ABA) in current-year needles of two different genotypes (AB-NSD-004 and AB-NSD-184) were monitored in branches collected from 20-year-old balsam fir (Abies balsamea L. (Mill.)) trees over a period of 11 months. A significant genotype × harvesting time interaction was observed for endogenous ABA levels and postharvest needle retention duration (NRD). A consistent pattern of seasonal variation in ABA concentration was observed in both genotypes, with the highest amount of ABA (7,887 ng g^?1 DW) accumulating in April and May. The highest levels of ABA coincided with the lowest postharvest NRD regardless of genotype. Nevertheless, genotypes differed in their ABA concentrations. Branches of genotype AB-NSD-184 sampled during August exhibited 170 days of NRD whereas those collected in May and June registered the lowest NRD of around 40 days. There was a significant negative correlation (P < 0.05) between endogenous ABA concentrations and postharvest NRD in genotype AB-NSD-184. Also, an inverse relationship was observed between the average daily photoperiod and the postharvest NRD (R ² = 0.35; P = 0.000) in the same genotype. Together with average daily temperature, the R ² value for this correlation reached the highest (0.75; P ≤ 0.00). Genotypes differed in their physiological responses to environmental cues and thus differed in their postharvest qualities. Average daily photoperiod and maximum daily temperature are strongly linked to the postharvest NRD through modulating endogenous ABA concentration.     

Screening of terrestrial and freshwater halotolerant cyanobacteria for antifungal activities

Cyanobacterial cultures tolerating 200 mmol l⁻¹ sodium chloride isolated from terrestrial and freshwater habitats of North Maharashtra region of India were evaluated for antifungal activity. Aqueous, methanol, n-propanol, and petroleum ether extracts of 40 cyanobacterial isolates belonging to nine genera were examined for inhibitory activity against five fungal pathogens. Eighteen isolates belonging to genus Oscillatoria dominated the population of halotolerant cyanobacterial cultures. Four antifungal bioassays viz. double layer agar method, disc diffusion assay, silica gel method, and minimum inhibitory concentration (MIC) were used to screen the cultures for antifungal activity. Among the solvents used, methanol extracts showed 34.9% inhibition followed by n-propanol, petroleum ether, and water exhibiting 30.2%, 18.6% and 16.2% inhibition, respectively. The double agar layer method was found to be a suitable method in preliminary screening for handling large number of cultures without extraction of compounds. However, in later screening experiments, silica gel method was seen to be advantageous over MIC and agar disc diffusion methods.