Green synthesis of isoamyl acetate via silica immobilized novel thermophilic lipase from <Emphasis Type="Italic">Bacillus aerius</Emphasis>

Isoamyl acetate, a pear or banana flavor, is widely used in food, beverage, cosmetic, and pharmaceutical industries. In the present work, lipase from Bacillus aerius was immobilized on silica gel matrix in the presence of a cross-linking agent, glutaraldehyde, and its efficiency in synthesizing isoamyl acetate using esterification reaction was studied. The esterification of acetic acid and isoamyl alcohol by silica-bound lipase was studied as a function of time and temperatures. The incubation time of 10 h, temperature of 55°C, substrate molar ratio 1: 1, and the amount of lipase as 1% were found to be optimal for the esterification reaction. The bound lipase catalyzed the esterification of acetic acid by isoamyl alcohol with the yield of about 68% under the optimized reaction conditions. The product was identified as isoamyl acetate using gas-liquid chromatography, nuclear magnetic resonance, and Fourier transform IR spectroscopy analysis by the presence of an ester group at the wavenumber of 1720.5 cm^–1.     

Outbreak of Abortions and Infertility in Thoroughbred Mares Associated with Waterborne <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

At a thoroughbred equine breeding farm near Hissar (Haryana), three mares aborted in their seventh month of pregnancy. The vaginal swabs of all aborted mares, and stomach contents, heart blood, liver, spleen and placenta of aborted fetuses yielded pure culture of Aeromonas hydrophila. In addition, A. hydrophila was also isolated from the vaginal swabs of three repeat breeding mares and faecal sample of a diarrheic foal. The source of infection was possibly water supply as all the water samples collected from taps, mother tank and storage tank were found to be positive for A. hydrophila. The antibiogram of all the isolates was similar showing resistance to ampicillin, carbenicillin, gentamicin, kanamycin and amikacin but sensitive to chloramphenicol, ciprofloxacin, cefuroxime, ceftriaxone, cotrimoxazole, cotrimazine, nitrofurantoin, streptomycin and tetracycline. All the 20 sera samples collected from three aborted and three repeat breeding, and eight in-contact mares, a diarrheic foal, three cows and two male buffaloes maintained at the same farm contained antibodies against A. hydrophila with titres ranging from 80 to 640. The water supply was instantly chlorinated using 0.05% sodium hypochlorite for three consecutive days and all the culturally positive mares were treated with intravaginal administration of 1 g ciprofloxacin, while the foal was given nitrofurantoin for three days. After one month, A. hydrophila could not be isolated either from mares or from their environment and antibody titre in all the seropositive animals showed a declining trend. Later, all the aborted and repeat breeding mares were confirmed to be pregnant. Thus, the present study indicated that water-borne A. hydrophila might be associated with equine abortions and infertility, and diarrhea in newborn foals.     

Ligand migration and hexacoordination in type 1 non-symbiotic rice hemoglobin

Type 1 non-symbiotic rice hemoglobin (rHb1) shows bis-histidyl heme hexacoordination and is capable of binding diatomic ligands reversibly. The biological function is as yet unclear, but the high oxygen affinity makes it unlikely to be involved in oxygen transport. In order to gain insight into possible physiological roles, we have studied CO rebinding kinetics after laser flash photolysis of rHb1 in solution and encapsulated in silica gel. CO rebinding to wt rHb1 in solution occurs through a fast geminate phase with no sign of rebinding from internal docking sites. Encapsulation in silica gel enhances migration to internal cavities. Site-directed mutagenesis of FB10, a residue known to have a key role in the regulation of hexacoordination and ligand affinity, resulted in substantial effects on the rebinding kinetics, partly inhibiting ligand exit to the solvent, enhancing geminate rebinding and enabling ligand migration within the internal cavities. The mutation of HE7, one of the histidyl residues involved in the hexacoordination, prevents hexacoordination, as expected, but also exposes ligand migration through a complex system of cavities. This article is part of a Special Issue entitled: Protein Dynamics: Experimental and Computational Approaches.     

Capacity optimization and scheduling of a multiproduct manufacturing facility for biotech products

A general mathematical framework has been proposed in this work for scheduling of a multiproduct and multipurpose facility involving manufacturing of biotech products. The specific problem involves several batch operations occurring in multiple units involving fixed processing time, unlimited storage policy, transition times, shared units, and deterministic and fixed data in the given time horizon. The different batch operations are modeled using state‐task network representation. Two different mathematical formulations are proposed based on discrete‐ and continuous‐time representations leading to a mixed‐integer linear programming model which is solved using General Algebraic Modeling System software. A case study based on a real facility is presented to illustrate the potential and applicability of the proposed models. The continuous‐time model required less number of events and has a smaller problem size compared to the discrete‐time model. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1221–1230, 2014     

Cytodiagnosis of eumycotic mycetoma: a case report

BACKGROUND: Mycetoma is a late-stage clinical manifestation of a subcutaneous infection produced by bacteria (actinomycetoma) or fungi (eumycetoma). Only a few articles have described the morphologic appearance of this uncommon pathology on cytology. The distinction between eumycetoma and actinomycetoma in fine needle aspiration cytology (FNAC) is as accurate as in histopathology, as demonstrated in the present case. CASE: A 30-year-old man presented with a large swelling on his left foot with a discharging sinus. FNAC of the swelling yielded pus-like material. Initial Papanicolaou and Giemsa stains showed the presence of septate, branching fungal hyphae and black granules against the inflammatory background. The presence of fungus was confirmed by PAS stain. The species was identified as Exophiala jeanselmei on fungal culture. CONCLUSION: Mycotic mycetoma can be accurately diagnosed by FNAC, which is a simple, inexpensive and rapid technique when there is a high index of suspicion. Special stains and culture studies are helpful in confirmation of diagnosis and species identification.     

Low plasma albumin levels are associated with increased plasma protein glycation and HbA1c in diabetes

Albumin is one of the most abundant plasma proteins and is heavily glycated in diabetes. In this study, we have addressed whether variation in the albumin levels influence glycation of plasma proteins and HbA1c. The study was performed in three systems: (1) streptozotocin (STZ)-induced diabetic mice plasma, (2) diabetic clinical plasma, and (3) in vitro glycated plasma. Diabetic mice and clinical plasma samples were categorized as diabetic high albumin plasma (DHAP) and diabetic low albumin plasma (DLAP) on the basis of their albumin levels. For the in vitro experiment, two albumin levels, high albumin plasma (HAP) and low albumin plasma (LAP), were created by differential depletion of plasma albumin. Protein glycation was studied by using a combination of two-dimensional electrophoresis (2DE), Western blotting, and LC-MS(E). In both mice and clinical experiments, an increased plasma protein glycation was observed in DLAP than in DHAP. Additionally, plasma albumin levels were negatively correlated with HbA1c. The in vitro experiment with differential depletion of albumin mechanistically showed that the low albumin levels are associated with increased plasma protein glycation and that albumin competes for glycation with other plasma proteins.     

Lewis‐Adduct Mediated Grain‐Boundary Functionalization for Efficient Ideal‐Bandgap Perovskite Solar Cells with Superior Stability

State‐of‐the‐art perovskite solar cells (PSCs) have bandgaps that are invariably larger than 1.45 eV, which limits their theoretically attainable power conversion efficiency. The emergent mixed‐(Pb, Sn) perovskites with bandgaps of 1.2–1.3 eV are ideal for single‐junction solar cells according to the Shockley–Queisser limit, and they have the potential to deliver higher efficiency. Nevertheless, the high chemical activity of Sn(II) in these perovskites makes it extremely challenging to control their physical properties and chemical stability, thereby leading to PSCs with relatively low PCE and stability. In this work, the authors employ the Lewis‐adduct SnF₂·3FACl additive in the solution‐processing of ideal‐bandgap halide perovskites (IBHPs), and prepare uniform large‐grain perovskite thin films containing continuously functionalized grain boundaries with the stable SnF₂ phase. Such Sn(II)‐rich grain‐boundary networks significantly enhance the physical properties and chemical stability of the IBHP thin films. Based on this approach, PSCs with an ideal bandgap of 1.3 eV are fabricated with a promising efficiency of 15.8%, as well as enhanced stability. The concept of Lewis‐adduct‐mediated grain‐boundary functionalization in IBHPs presented here points to a new chemical route for approaching the Shockley–Queisser limit in future stable PSCs.     

Evaluation of potential flavonoid inhibitors of glyoxalase-I based on virtual screening and in vitro studies

Glyoxalase-I (GLO-I) is a component of the ubiquitous detoxification system involved in the conversion of methylglyoxal (MG) to d-lactate in the glycolytic pathway. MG toxicity arises from its ability to form advanced glycation end products. GLO-I has been reported to be frequently overexpressed in various types of cancer cells. In this study, we performed structure-based virtual screening of focused flavonoids commercial library to identify potential and specific inhibitors of GLO-I. The compounds were ranked based on Glide extra precision docking score and five hits (curcumin, quercetin, morin, naringin and silibinin) were selected on the basis of their interaction with active site amino acid residues of GLO-I. Mixed mode QM/MM calculation was performed on the top-scoring hit to ascertain the role of zinc ion in ligand binding. In addition, the identified hits were subjected to MM/GBSA binding energy prediction, ADME prediction and similarity studies. The hits were tested in vitro for cell viability, and GLO-I inhibition. Naringin (ST072162) was found to be most potent inhibitor of GLO-I among the identified hits with highest glide XP dock score of ?14.906. These findings suggest that naringin could be a new scaffold for designing inhibitors against GLO-I with potential application as anticancer agents.