Continuous production of galacto-oligosaccharides from lactose using immobilized β-galactosidase from Bacillus circulans

Summary -Galactosidase-2 (-d-galactoside galactohydrolase, EC 3.2.1.23) from Bacillus circulans was purified using hydroxyapatite gel chromatography and immobilized onto Duolite ES-762 (phenolformaldehyde resin) and Merckogel (controlled pore silica gel) for continuous production of galacto-oligosaccharides using lactose as the substrate. The maximum amount of ologosaccharides produced by the immobilized enzyme was 35–40% of the total sugar during hydrolysis of 4.56% lactose. Partially purified -galactosidase from B. circulans was also immobilized onto various supports for the same purpose. The stability of the immobilized -galactosidase-2 or partially purified enzyme during a continuous reaction depended on their supports and specific activity. Of the supports tested, Merckogel was best for operational stability. With this support, the enzyme was quite stable with specific activity up to 15 units/g of wet gel; it was reversibly inactivated with more.     

Slow development of ALS-like spinal cord pathology in mutant valosin-containing protein gene knock-in mice

Pathological features of amyotrophic lateral sclerosis (ALS) include, in addition to selective motor neuron (MN) degeneration, the occurrence of protein aggregates, mitochondrial dysfunction and astrogliosis. SOD1 mutations cause rare familial forms of ALS and have provided the most widely studied animal models. Relatively recent studies implicating another protein, TDP-43, in familial and sporadic forms of ALS have led to the development of new animal models. More recently, mutations in the valosin-containing protein (VCP) gene linked to the human genetic disease, Inclusion Body Myopathy associated with Paget''s disease of bone and frontotemporal dementia (IBMPFD), were found also to be associated with ALS in some patients. A heterozygous knock-in VCP mouse model of IBMPFD (VCP^R155H/+) exhibited muscle, bone and brain pathology characteristic of the human disease. We have undertaken studies of spinal cord pathology in VCP^R155H/+ mice and find age-dependent degeneration of ventral horn MNs, TDP-43-positive cytosolic inclusions, mitochondrial aggregation and progressive astrogliosis. Aged animals (∼24–27 months) show electromyography evidence of denervation consistent with the observed MN loss. Although these animals do not develop rapidly progressive fatal ALS-like disease during their lifespans, they recapitulate key pathological features of both human disease and other animal models of ALS, and may provide a valuable new model for studying events preceding onset of catastrophic disease.     

Microbial rRNA gene expression and co‐occurrence profiles associate with biokinetics and elemental composition in full‐scale anaerobic digesters

This study examined whether the abundance and expression of microbial 16S rRNA genes were associated with elemental concentrations and substrate conversion biokinetics in 20 full‐scale anaerobic digesters, including seven municipal sewage sludge (SS) digesters and 13 industrial codigesters. SS digester contents had higher methane production rates from acetate, propionate and phenyl acetate compared to industrial codigesters. SS digesters and industrial codigesters were distinctly clustered based on their elemental concentrations, with higher concentrations of NH₃‐N, Cl, K and Na observed in codigesters. Amplicon sequencing of 16S rRNA genes and reverse‐transcribed 16S rRNA revealed divergent grouping of microbial communities between mesophilic SS digesters, mesophilic codigesters and thermophilic digesters. Higher intradigester distances between Archaea 16S rRNA and rRNA gene profiles were observed in mesophilic codigesters, which also had the lowest acetate utilization biokinetics. Constrained ordination showed that microbial rRNA and rRNA gene profiles were significantly associated with maximum methane production rates from acetate, propionate, oleate and phenyl acetate, as well as concentrations of NH₃‐N, Fe, S, Mo and Ni. A co‐occurrence network of rRNA gene expression confirmed the three main clusters of anaerobic digester communities based on active populations. Syntrophic and methanogenic taxa were highly represented within the subnetworks, indicating that obligate energy‐sharing partnerships play critical roles in stabilizing the digester microbiome. Overall, these results provide new evidence showing that different feed substrates associate with different micronutrient compositions in anaerobic digesters, which in turn may influence microbial abundance, activity and function.     

Multi-domain network infrastructure based on P4 programmable devices for Digital Data Marketplaces

Cluster Computing - There are many organizations interested in sharing data with others, and they can do this only if a multi-domain secure platform is available. Such platforms, often referred to...     

Change in turnover capacity of crude recombinant dye-decolorizing peroxidase (rDyP) in batch and fed-batch decolorization of Remazol Brilliant Blue R

Decolorization of the representative anthraquinone dye, Remazol Brilliant Blue R (RBBR) was assessed to determine the practical potential of crude recombinant dye-decolorizing peroxidase generated by Aspergillus oryzae (rDyP) in term of turnover capacity of rDyP. The turnover capacity, defined as the milligram of RBBR decolorized per unit of rDyP inactivated over the catalytic life time of rDyP, was quantified under condition by H₂O₂ -mediated rDyP inactivation. In batch culture, equimolar batch addition of H₂O₂ and RBBR yielded complete decolorization of RBBR by rDyP, with a turnover capacity of 4.75. In stepwise fed-batch addition of H₂O₂, the turnover capacity increased to 5.76, and by increasing dye concentration, it reached 14.3. When H₂O₂ was added in continuous fed-batch to minimize rDyP inactivation and 1.6 mM dye was added in stepwise fed-batch mode, the turnover capacity increased to 20.4. At this turnover capacity, 1 l of crude rDyP solution containing 5,000 U could decolorize up to 102 g RBBR in 650 min.     

Functional Characterization of CaVα2δ Mutations Associated with Sudden Cardiac Death

L-type Ca²⁺ channels play a critical role in cardiac rhythmicity. These ion channels are oligomeric complexes formed by the pore-forming Ca_Vα1 with the auxiliary Ca_Vβ and Ca_Vα2δ subunits. Ca_Vα2δ increases the peak current density and improves the voltage-dependent activation gating of Ca_V1.2 channels without increasing the surface expression of the Ca_Vα1 subunit. The functional impact of genetic variants of CACNA2D1 (the gene encoding for Ca_Vα2δ), associated with shorter repolarization QT intervals (the time interval between the Q and the T waves on the cardiac electrocardiogram), was investigated after recombinant expression of the full complement of L-type Ca_V1.2 subunits in human embryonic kidney 293 cells. By performing side-by-side high resolution flow cytometry assays and whole-cell patch clamp recordings, we revealed that the surface density of the Ca_Vα2δ wild-type protein correlates with the peak current density. Furthermore, the cell surface density of Ca_Vα2δ mutants S755T, Q917H, and S956T was not significantly different from the cell surface density of the Ca_Vα2δ wild-type protein expressed under the same conditions. In contrast, the cell surface expression of Ca_Vα2δ D550Y, Ca_Vα2δ S709N, and the double mutant D550Y/Q917H was reduced, respectively, by ≈30–33% for the single mutants and by 60% for the latter. The cell surface density of D550Y/Q917H was more significantly impaired than protein stability, suggesting that surface trafficking of Ca_Vα2δ was disrupted by the double mutation. Co-expression with D550Y/Q917H significantly decreased Ca_V1.2 currents as compared with results obtained with Ca_Vα2δ wild type. It is concluded that D550Y/Q917H reduced inward Ca²⁺ currents through a defect in the cell surface trafficking of Ca_Vα2δ. Altogether, our results provide novel insight in the molecular mechanism underlying the modulation of Ca_V1.2 currents by Ca_Vα2δ.     

Evaluation of Crocus sativus L. (saffron) on male erectile dysfunction: A pilot study

In this study, the effect of Crocus sativus (saffron) was studied on male erectile dysfunction (ED). Twenty male patients with ED were followed for ten days in which each morning they took a tablet containing 200 mg of saffron. Patients underwent the nocturnal penile tumescence (NPT) test and the international index of erectile function questionnaire (IIEF-15) at the start of the treatment and at the end of the ten days. After the ten days of taking saffron there was a statistically significant improvement in tip rigidity and tip tumescence as well as base rigidity and base tumescence. ILEF-15 total scores were significantly higher in patients after saffron treatment (before treatment 22.15±1.44; after treatment 39.20±1.90, p<0.001). Saffron showed a positive effect on sexual function with increased number and duration of erectile events seen in patients with ED even only after taking it for ten days.     

Isolation of copper oxide (CuO) nanoparticles resistant Pseudomonas strains from soil and investigation on possible mechanism for resistance

The present study deals with isolation and characterization of copper oxide nanoparticles resistant Pseudomonas strains that were isolated from the soil collected from mining and refining sites of Sarcheshmeh copper mine in the Kerman Province of Iran. The three isolates were selected based on high level of copper oxide nanoparticles (CuO NPs) resistance. The isolates were authentically identified as Pseudomonas fluorescens CuO-1, Pseudomonas fluorescens CuO-2 and Pseudomonas sp. CuO-3 by morphological, biochemical and 16S rDNA gene sequencing analysis. The growth pattern of these isolates with all the studied CuO NPs concentrations was similar to that of control (without CuO NPs) indicating that CuO NPs would not affect the growth of isolated strains. A reduction in the amount of exopolysaccharides was observed after CuO NPs—P. fluorescens CuO-1 culture supernatant interaction. The Fourier transform infrared spectroscopy (FT-IR) peaks for the exopolysaccharides extracted from the bacterial culture supernatant and the interacted CuO NPs were almost similar. The exopolysaccharide capping of the CuO NPs was confirmed by FT-IR and X-ray diffraction analysis. The study of bacterial exopolysaccharides capped CuO NPs with E. coli PTCC 1338 and S. aureus PTCC 1113 showed less toxicity compared to uncoated CuO NPs. Our study suggests that the capping of nanoparticles by bacterially produced exopolysaccharides serve as the probable mechanism of tolerance.