Expression of Aspergillus nidulans phy Gene in Nicotiana benthamiana Produces Active Phytase with Broad Specificities

A full-length phytase gene (phy) of Aspergillus nidulans was amplified from the cDNA library by polymerase chain reaction (PCR), and it was introduced into a bacterial expression vector, pET-28a. The recombinant protein (rPhy-E, 56 kDa) was overexpressed in the insoluble fraction of Escherichia coli culture, purified by Ni-NTA resin under denaturing conditions and injected into rats as an immunogen. To express A. nidulans phytase in a plant, the full-length of phy was cloned into a plant expression binary vector, pPZP212. The resultant construct was tested for its transient expression by Agrobacterium-infiltration into Nicotiana benthamiana leaves. Compared with a control, the agro-infiltrated leaf tissues showed the presence of phy mRNA and its high expression level in N. benthamiana. The recombinant phytase (rPhy-P, 62 kDa) was strongly reacted with the polyclonal antibody against the nonglycosylated rPhy-E. The rPhy-P showed glycosylation, two pH optima (pH 4.5 and pH 5.5), an optimum temperature at 45~55 °C, thermostability and broad substrate specificities. After deglycosylation by peptide-N-glycosidase F (PNGase-F), the rPhy-P significantly lost the phytase activity and retained 1/9 of the original activity after 10 min of incubation at 45 °C. Therefore, the deglycosylation caused a significant reduction in enzyme thermostability. In animal experiments, oral administration of the rPhy-P at 1500 U/kg body weight/day for seven days caused a significant reduction of phosphorus excretion by 16% in rat feces. Besides, the rPhy-P did not result in any toxicological changes and clinical signs.     

Polyaniline–13X zeolite composite‐supported platinum electrocatalysts for direct methanol fuel cell applications

We successfully synthesized 13X zeolite using a hydrothermal method. Then, composites of polyaniline (PANI) with 13X zeolite and PANI–13X with platinum were prepared by chemical oxidative polymerization and chemical reduction, respectively. Field emission scanning electron microscopy, X‐ray diffraction, Raman spectroscopy and Brunauer–Emmett–Teller techniques were used to characterize the PANI–Pt and PANI–Pt–13X composites. Further, the electrocatalytic activity towards methanol oxidation of the synthesized catalysts was explored using cyclic voltammetry in 1 mol L^?1 CH₃OH + 0.5 mol L^?1 H₂SO₄ solution. From the obtained results, PANI–Pt–13X shows superior performance compared to PANI–Pt towards methanol oxidation and electrical conductivity. Hence, the 13X zeolite‐incorporated PANI–Pt composite could be an efficient catalyst for direct methanol fuel cell applications. © 2019 Society of Chemical Industry     

Fabrication, characterization and invitro bioactivity evaluation of QPSU/TiO2 composite membranes

Quaternized Polysulfone (QPSU) is a widely investigated material in the industry because of its unique properties such as resistance to corrosion and high mechanical properties. The ionic nature of the compound can be exploited for medical applications such as in haemodialysis, drug delivery and tissue engineering. In this study, composite membranes of QPSU with varying concentrations of Titanium oxide (TiO₂) were prepared and characterized using FT-IR, ¹H-NMR, XRD, TGA and SEM. The bioactivity of the membranes was studied by immersing them in simulated body fluid (SBF) for 7 days and subsequently observing under SEM for the formation of calcium-phosphate (Ca–PO₄) layer on the surface of the membranes. The formation of Ca–PO₄ on the samples was confirmed using FT-IR and EDAX. The results were compared with those obtained for QPSU membranes and the effect of TiO₂ concentration on the membrane properties was analyzed. It was observed that the percentage crystallinity of the composites increased upto a filler concentration of 5 wt% beyond which it decreased. TGA studies revealed an increase in the thermal stability of the composites with increasing filler concentrations. While optimum bioactivity was observed in the samples containing 5 wt% of TiO₂, higher filler content resulted in the formation of denser calcium—phosphate layer on the surface of the composites. The study shows that quaternized polysulphone/TiO₂ composites are promising bio composites having great potential for application in health care.     

Hypoglycaemic and Hypolipidaemic Effects of Withania somnifera Root and Leaf Extracts on Alloxan-Induced Diabetic Rats

Withania somnifera is an important medicinal plant, which is used in traditional medicine to cure many diseases. Flavonoids were determined in the extracts of W. somnifera root (WSREt) and leaf (WSLEt). The amounts of total flavonoids found in WSREt and WSLEt were 530 and 520 mg/100 g dry weight (DW), respectively. Hypoglycaemic and hypolipidaemic effects of WSREt and WSLEt were also investigated in alloxan-induced diabetic rats. WSREt and WSLEt and the standard drug glibenclamide were orally administered daily to diabetic rats for eight weeks. After the treatment period, urine sugar, blood glucose, haemoglobin (Hb), glycosylated haemoglobin (HbA1C), liver glycogen, serum and tissues lipids, serum and tissues proteins, liver glucose-6-phosphatase (G6P) and serum enzymes like aspartate transaminase (AST), alanine transaminase (ALT), acid phosphatase (ACP) and alkaline phosphatase (ALP) levels were determined. The levels of urine sugar, blood glucose, HbA1C, G6P, AST, ALT, ACP, ALP, serum lipids except high density lipoprotein-bound cholesterol (HDL-c) and tissues like liver, kidney and heart lipids were significantly (p < 0.05) increased, however Hb, total protein, albumin, albumin:globulin (A:G) ratio, tissues protein and glycogen were significantly (p < 0.05) decreased in alloxan-induced diabetic rats. Treatment of the diabetic rats with WSREt, WSLEt and glibenclamide restored the changes of the above parameters to their normal level after eight weeks of treatment, indicating that WSREt and WSLEt possess hypoglycaemic and hypolipidaemic activities in alloxan-induced diabetes mellitus (DM) rats.     

Down syndrome associated malformations

This paper reports the associated malformations and the clinical findings that were observed in 417 cytogenetically confirmed Down Syndrome patients. Among them congenital heart defects have occurred more frequently [75; 17.98%] than osteoarticular malformations [23; 5.52]; eye anomalies [22; 5.27%]; and gastroenterological malformations [n 16; 3.84%]. With regard to prognosis and treatment appropriate counselling has been given to Down Syndrome patients and their families.     

Characterization of paper mill sludge as a renewable feedstock for sustainable hydrogen and biofuels production

Paper and pulp mills generate substantial quantities of cellulose-rich sludge materials that are disposed in landfills at a large scale. For sustainability purposes, sludge materials can be bioprocessed to produce renewable fuels and useful chemicals. The enzymatic hydrolysis of cellulose is the process bottleneck that affects the conversion economics directly by using zero-cost raw materials. In order to study and optimize the process, the characteristics of the sludge raw materials should be first evaluated. In this work, sludge samples were obtained from paper mills located at different locations in Wisconsin and Minnesota. Part of the sludge samples was washed (de-ashed) with hydrochloric acid while the other part remained unwashed. The samples were subjected to multiple spectroscopic analyses techniques to evaluate the morphological properties of cellulose fibers and to estimate the total structural carbohydrate content. The results showed that the de-ashing process changed some fiber characteristics and cellulose crystallinity structure in all sludge samples. Sludge sample A (obtained from Kraft pulp and recycled paper mill region) showed a high percentage of fiber, with crystalline cellulose, compared to the other two sludge samples suggesting that sludge A is a valuable source to make value-added products. Aspen Plus mass and energy calculations performed in view of the ‘zero’ cost and the reliable supply of sludge raw materials producing 2 mol H₂/mol glucose. Moreover, the results showed that extracting crystalline cellulose from these sludge samples is more profitable than crystalline cellulose made from the other lignocellulosic feedstocks. The results reported here showed that the utilization of these sludge materials would be an economically attractive and promising alternative for the production of hydrogen.     

Techno-economic analysis and a novel assessment technique of paper mill sludge conversion to bioethanol toward sustainable energy production

A novel bioprocess design to convert paper mill sludge (PMS) to biofuels is proposed in this work. The design utilizes cellulosic fiber recovered from the PMS via optimized de-ashing (HCl washing) step. This work specifically provided a technical and economic analysis of paper mill sludge conversion into biofuel production using a novel protocol. The protocol is based on scanning electron microscopy (SEM) analysis to assess the quality of the contained cellulose prior to further processing. The results are crucially important to determine the suitability of the PMS feedstock to produce biofuels. SEM analysis was employed as a preliminary screening tool to evaluate sludge digestibility and conversion. The SEM characterization technique established a direct relationship between the fiber morphology, presence of crystals salts and sugar yield after enzymatic hydrolysis. Substantial structural changes were observed before and after de-ashing the sludge samples, leading to a correlation between the surface morphology and the washing step. The results suggested that de-ashing changes the surface morphology and upon analysis, increased the sugar yield up to about 86% as opposed to 2.2% in sludge sample A as an example. The PMS conversion into biofuel was simulated using Aspen PLUS and compared to a similar process using corn stover as feedstock. The simulation results showed it is 20% cheaper to produce bioethanol from PMS compared to corn stover. The simulation revealed less energy demand by around 13 320 MJ/h compared to that when corn stover was used.     

Dry colour prediction of leather from its wet state

The present investigation describes the development of a regression model for dry colour prediction based on the colour of wet leather. Such a prediction will be useful in the colour matching of leather – the time delay in drying of wet leather and determination of the final colour can be avoided. The use of computer technology interfaced with a reflectance spectrophotometer provides a novel approach to the prediction of dry colour from wet colour. In this study, five different regression models, i.e. linear fit, log fit, quadratic fit, log quadratic fit and cubic fit, were developed, with the dry reflectance of the leather sample as a dependent variable and the wet reflectance of the leather as an independent variable. The relationship between moisture content and surface leather colour, a prerequisite for enabling prediction, was studied. The obtained log quadratic model can be used for accurate prediction of dry colour from wet colour.     

Carbon supported silver (Ag/C) electrocatalysts for alkaline membrane fuel cells

This study reports on the performance of activated carbon supported silver catalyst (Ag/C) as electrocatalyst in an alkaline membrane fuel cell (AMFC), using alkalized poly (styrene ethylene butylene poly styrene) [APSEBS] as the electrolyte membrane. Carbon supported silver catalyst (Ag/C) with different metal loading was synthesized by means of wet impregnation method. The prepared electrocatalyst was characterized by X-ray diffraction pattern (XRD), thermogravimetric analysis (TGA), UV–Visible diffuse reflectance spectra (DRS-UV) and Raman spectroscopy. Surface morphology analysis of the prepared electrocatalyst using scanning electron microscopy (SEM) revealed the heterogeneous distribution of Ag on carbon support. The performance of the prepared electrocatalyst was evaluated with a home made AMFC using a novel anion exchange membrane (APSEBS). A maximum cell voltage and power density of 0.69 V and 109 mW/cm², respectively, was achieved at 60 °C for the home made 10 wt% (Ag/C) cathode catalyst and anion exchange membrane. Further, the prepared electrocatalyst was subjected to cyclic voltammetry studies to evaluate the methanol oxidation for Direct methanol alkaline fuel cell (DMAFC) applications.     

The telemetry system

This paper describes the telemetry system employed inAryabhata. The telemetry link provides a means for monitoring diagnostic and other parameters for efficient and controlled operation of the satellite besides transmitting data pertaining to the three scientific experiments. The design specifications and details, qualification tests and in-orbit performance of the telemetry system are also described in this paper.