Biochemistry, Synthesis, and Applications of Bacterial Cellulose: A Review.

The potential of cellulose nanocomposites in the new-generation super-performing nanomaterials is huge, primarily in medical and environment sectors, and secondarily in food, paper, and cosmetic sectors. Despite substantial illumination on the molecular aspects of cellulose synthesis, various process features, namely, cellular export of the nascent polysaccharide chain and arrangement of cellulose fibrils into a quasi-crystalline configuration, remain obscure. To unleash its full potential, current knowledge on nanocellulose dispersion and disintegration of the fibrillar network and the organic/polymer chemistry needs expansion. Bacterial cellulose biosynthesis mechanism for scaled-up production, namely, the kinetics, pathogenicity, production cost, and product quality/consistency remain poorly understood. The bottom-up bacterial cellulose synthesis approach makes it an interesting area for still wider and promising high-end applications, primarily due to the nanosynthesis mechanism involved and the purity of the cellulose. This study attempts to identify the knowledge gap and potential wider applications of bacterial cellulose and bacterial nanocellulose. This review also highlights the manufacture of bacterial cellulose through low-cost substrates, that is, mainly waste from brewing, agriculture, food, and sugar industries as well as textile, lignocellulosic biorefineries, and pulp mills.

Eosinophils Restrict Diesel Exhaust Particles-induced Cell Proliferation of Lung Epithelial A549 Cells via Interleukin-13 Mediated Mechanisms: Implications for Tissue Remodeling and Fibrosis.

BACKGROUND: Diesel exhaust particles (DEPs) affect lung physiology and cause serious damage to the lungs. A number of studies demonstrated that eosinophils play a very important role in the development of tissue remodeling and fibrosis of the lungs. However, the exact mechanism of pathogenesis of tissue remodeling and fibrosis is not known. METHODS: Both in vitro and in vivo models were used in the study. HL-60 and A549 cells were also utilized in the study. 8 to 12 weeks old BALB/c mice were used for the in vivo study. Cell viability by MTT assay and RNA isolation by tri reagent was accomplished. mRNA expression of inflammatory genes was accomplished by real-time PCR or qPCR. Immunohistochemistry was done to assess the localization and expressions of proteins. One-way ANOVA followed by a post hoc test was done for the statistical analysis. Graph-Pad prism 5 software was used for statistical analysis. RESULTS: For the first time, we demonstrate that interleukin-13 plays a very important role in the development of tissue remodeling and fibrosis. We report that diesel exhaust particles significantly induce eosinophils cell proliferation and interleukin-13 release in in vitro culture conditions. Supernatant collected from DEP-induced eosinophils cells significantly restricts cell proliferation of epithelial cells in response to exposure to diesel exhaust particles. Furthermore, purified interleukin-13 decreases the proliferation of A549 cells, highlighting the involvement of IL- 13 in tissue remodeling. Notably, Etoricoxib (selective COX-2 inhibitor) did not inhibit the DEPtriggered release of interleukin-13, suggesting another cell signaling pathway. The in vivo exposure of DEP to the lungs of mice resulted in a high level of eosinophils degranulation as depicted by the EPX-1 immunostaining and altered level of mRNA expressions of inflammatory genes. We also found that a-SMA, fibroblast specific protein (FSP-1), has been changed in response to DEP in the mice lungs along with the mediators of inflammation. CONCLUSION: Altogether, we elucidated the mechanistic role of eosinophils and IL-13 in the DEP-triggered proliferation of lungs cells, thus providing an insight into the pathophysiology of tissue remodeling and fibrosis of lungs.

Virus as a Nanocarrier for Drug Delivery Redefining Medical Therapeutics - A Status Report.

Over the last two decades, drug delivery systems have evolved at a tremendous pace. Synthetic nanoparticles have played an important role in vaccine design and delivery as these have shown improved safety and efficacy over conventional formulations. Nanocarriers formulated by natural, biological building blocks have become an important tool in biomedicine. A successful nanocarrier must possess specific properties like evading the host immune system, target specificity, cellular entry, escape from endosomes, and the ability to release the active material into the cytoplasm. The virus can perform some or all of these functions, making it a suitable candidate as a naturally occurring nanocarrier. Viruses could be made non-infectious and non-replicative without compromising their ability to penetrate cells, making them useful for a vast spectrum of applications. Currently, many such carrier molecules as bio-nanocapsules are at various development stages. This review covers the advances in the field of viruses as potential nanocarriers and discusses the related technologies and strategies to target specific cells by using virus-inspired nanocarriers. These virus-based nanocarriers could provide solutions to address pressing and emerging concerns in infectious diseases in the future.

Sub-fertility in crossbred bulls: Identification of proteomic alterations in spermatogenic cells using high throughput comparative proteomics approach.

The present study was carried out to compare the proteomic profiles of spermatogenic cells of crossbred and zebu cattle in an effort to understand the possible reasons for a higher incidence of sub-fertility in crossbred bulls. The spermatogenic cells collected from the testes of pre-pubertal (6 mo) and adult (24 mo) crossbred and zebu males through fine needle aspiration were proliferated in vitro, and proteomic profiling was done using a shotgun proteomics approach. The age- and species-specific variations in the expression level of proteins were identified in spermatogenic cells. The number of differentially expressed proteins (DEPs) identified in pre-pubertal zebu and crossbred was 546, while 579 DEPs were identified between adult zebu and crossbred bulls. Out of these, 194 DEPS were common to these groups and 40 DEPs displayed a fold change ≥2. However, only 20 proteins exhibited similar expression variation trends (upregulated or downregulated) among pre-pubertal as well as adult zebu and crossbred bulls. Out of these 20 DEPs, 13 proteins were upregulated, and 7 proteins were downregulated in spermatogenic cells of zebu compared to crossbred bulls. Among the upregulated proteins were RPLP2, PAXIP1, calumenin, prosaposin, GTF2F1, TMP2, ubiquitin conjugation factor E4A, COL1A2, vimentin, protein FAM13A, peripherin, GFPT2, and GRP78. Seven proteins that were downregulated in zebu bulls compared to crossbred included APOA1, G patch domain-containing protein 1, NAD P transhydrogenase mitochondrial, glutamyl aminopeptidase, synaptojanin 1 fragment, Arf GAP with SH3 domain ANK repeat and PH domain-containing protein 1, and protein transport protein sec16B. It was inferred that the proteins associated with sperm function and fertilization processes, such as calumenin, prosaposin, vimentin, GRP78, and APOA1 could be studied further to understand the precise cause of subfertility in crossbred bulls.

Human Cadaveric Donor Cornea Derived Extra Cellular Matrix Microparticles for Minimally Invasive Healing/Regeneration of Corneal Wounds.

Biological materials derived from extracellular matrix (ECM) proteins have garnered interest as their composition is very similar to that of native tissue. Herein, we report the use of human cornea derived decellularized ECM (dECM) microparticles dispersed in human fibrin sealant as an accessible therapeutic alternative for corneal anterior stromal reconstruction. dECM microparticles had good particle size distribution (≤10 µm) and retained the majority of corneal ECM components found in native tissue. Fibrin-dECM hydrogels exhibited compressive modulus of 70.83 ± 9.17 kPa matching that of native tissue, maximum burst pressure of 34.3 ± 3.7 kPa, and demonstrated a short crosslinking time of ~17 min. The fibrin-dECM hydrogels were found to be biodegradable, cytocompatible, non-mutagenic, non-sensitive, non-irritant, and supported the growth and maintained the phenotype of encapsulated human corneal stem cells (hCSCs) in vitro. In a rabbit model of anterior lamellar keratectomy, fibrin-dECM bio-adhesives promoted corneal re-epithelialization within 14 days, induced stromal tissue repair, and displayed integration with corneal tissues in vivo. Overall, our results suggest that the incorporation of cornea tissue-derived ECM microparticles in fibrin hydrogels is non-toxic, safe, and shows tremendous promise as a minimally invasive therapeutic approach for the treatment of superficial corneal epithelial wounds and anterior stromal injuries.

Response surface methodology based optimization and scale-up production of amylase from a novel bacterial strain, Bacillus aryabhattai KIIT BE-1.

A novel strain KIIT BE-1 isolated from a specialized environment, screened through starch iodine test from a set of eighty-five biodigestate isolates, produced amylase maximally when cultured for 48 h at 37 °C. The molecular and biochemical characterization confirmed it as a strain of Bacillus aryabhattai. It exhibited optimal amylase activity (3.20 U/ml) at 36 h post incubation with a media combination of starch and yeast extract for C-N source respectively. Statistical optimisation by response surface modeling showed R2 values of 0.9645 for biomass and 0.9831 for amylase activity, suggesting the significance of the model. The optimised medium (10.25 % starch, 5.0 % peptone, 5.18 % yeast extract, pH 7.3) enhanced the enzyme activity to 4.16 U/ml (1.39-fold) from 3.20 U/ml of un-optimised medium. Further, the biomass yield and the enzymatic activity in optimized medium and process conditions increased by 1.14 and 1.21 folds subjected to a 5 l scaled-up operation in a lab-scale bioreactor.

A reliable animal model of corneal stromal opacity: Development and validation using in vivo imaging.

PURPOSE: To validate an animal model of corneal stromal opacity by using objective vision-independent in vivo imaging metrics. METHODS: This was a prospective study, with two arms: (i) observational human arm which included 14 patients with healed unilateral ulcerative keratitis; and (ii) experimental rabbit arm, which included 6 New Zealand white rabbits. A 3-mm central wound was created in the left eye of the rabbits by manually removing 200-250 µm of the superficial stroma, followed by rotating-burr application. Both groups underwent photography, high-resolution anterior segment optical coherence tomography, and Scheimpflug imaging using similar diagnostic platforms and standardized image capturing protocols. Parameters studied were relative change in (i) corneal thickness; (ii) corneal epithelial: stromal (E:S) reflectivity ratio; (iii) corneal stromal light scattering using densitometry; and (iv) central corneal keratometry. RESULTS: In the experimental arm, there was a significant decrease in corneal thickness (273 ± 51.3 vs. 407.3 ± 10.3 µm, p = 0.0038), E:S reflectivity ratio (0.71 ± 0.09 vs. 0.99 ± 0.06, p = 0.0018), and keratometry (40.4 ± 2.3 vs. 45.8 ± 0.9D, p = 0.0033) and increase in densitometry (54.2 ± 11.65 vs.18.7 ± 3.8 GSU, p = 0.0001) from baseline, which stabilized at 4 to 8-weeks post-wounding (p > 0.3632). At 8-weeks, the relative change from baseline in corneal thickness (28.4 ± 13.5% vs.22.4 ± 13%, p = 0.368), E:S reflectivity ratio (28.1 ± 11.5% vs. 30.6 ± 8.9%, p = 0.603), corneal densitometry (204.17 ± 97.3% vs. 304.9 ± 113.6%, p = 0.1113), and central corneal keratometry (13.6 ± 6.9% vs. 18.9 ± 7.4%, p = 0.1738) in rabbits was similar to human corneal scars. CONCLUSION: The animal model of corneal opacification was objectively comparable to human post-keratitis scars and can be valuable for in vivo evaluation of emerging therapies for corneal opacities.

Differential seminal plasma proteome signatures of acute lymphoblastic leukemia survivors.

With advances in therapeutic methods, there is a high survival rate among leukemia patients, of an extent more than 80%. However, chemotherapeutic drugs used to treat these patients have adverse effects on their overall health profile including fertility. The primary aim of this study was to identify differentially expressed proteins in seminal plasma of acute lymphoblastic leukemia (ALL) survivors compared to age-matched healthy controls, which can provide molecular basis of idiopathic infertility in such survivors. Differential proteome profiling was performed by 2D-differential in-gel electrophoresis, protein spots were identified by mass spectrometry and selective differentially expressed proteins (DEPs) were validated by western blotting and ELISA method. Out of eight DEPs identified, five proteins (isocitrate dehydrogenase 1, semenogelin 1, lactoferrin, prolactin-inducible protein, and human serum albumin) were upregulated and three (pepsinogen, prostate specific antigen and prostatic acid phosphatase) were downregulated. Expression profiles of these proteins are suggestive of reduction in semen quality in ALL survivors and can further be explored to determine their fertility status.

Intrinsic molecular insights to enhancement of biogas production from kitchen refuse using alkaline-microwave pretreatment.

The current study analyzed and optimized the concentration of NaOH for alkaline pretreatment of kitchen refuse for biogas production. Also, the benefits of microwave assistance in enhanced biogasification of kitchen refuse were evaluated. The TS, VS and structural changes were compared using standard experimental techniques. Molecular dynamics was investigated for the molecular level changes leading to higher biogasification in NaOHmicrowave combined pretreatment. The methane and biogas yields were calculated to validate the benefits of microwave assistance in efficient biogasification. The NaOH-microwave combined pretreatment showed higher VS production. Microwave treatment degraded and removed lignin more efficiently. Molecular dynamics studies revealed the induction of configurational instability in lignin and cellulose molecules with variable temperatures. The methane and biogas production increased with 6% NaOH concentration, and decreased at higher NaOH concentration till 10%. Microwave assistance declined the required NaOH concentration further to 4%. Thus, as compared to 6% NaOH concentration required for an efficient pretreatment, the kitchen refuse was efficiently pretreated with 4% NaOH concentration when combined with a 30 min duration microwaving. The experimental and computational data provided a detailed analysis proposing an optimized, novel and promising method to pretreat kitchen refuse for efficient and enhanced biogas production.

A comparative study of biogasification of wheat straw, sugarcane bagasse and pressmud.

A study to compare biogas production potentials of wheat straw, sugarcane bagasse and pressmud was conducted at pH 8.0, temperature 40 °C and substrate concentration 20 g/L. Raw substrates were thermogravimetrically and Fourier-transform infrared spectroscopically characterised. TGA showed the weight loss of samples attributable to moisture, hemicellulose, cellulose and lignin losses. FTIR analysis indicated functional groups characteristics of hemicellulose, cellulose and lignin. Biogas production was the maximum between 10th and 25th day for all the tests. WS with 10% inoculum showed the highest cumulative biogas production of 370 mL/g followed by the SB (316 mL/g) and PM (211 mL/g) counterparts. The corresponding values with 5% inoculum were 303 mL/g (WS), 244 mL/g (SB) and 152 mL/g (PM). The inoculum volume also positively affected the cumulative biogas production (22.1, 29.5 and 38.8% respectively). The higher volatile fatty acids as observed in case of WS which further facilitated higher biogas production could be due to its maximum volatile solids content (88.9%) and water swelling capacity (7.37). A consistently increasing trend in the methane content (varying between 54 and 61%) in all the tests was observed till the 20th day. The biogas (7.7-21.7 mL/g) and the methane (35-42%) contents showed a decreasing trend thereafter, the lowest being observed during the 35-40-day period.

Performance evaluation of vinasse treatment plant integrated with physico-chemical methods.

With an objective to assess environmental management criteria of a vinasse treatment plant (VTP) and to evaluate the critical environmental parameters, a study was undertaken in a multi-product (packaged apple juice, distillery, brewery, packaged drinking water) brewery-cum-distillery unit. The facility with a volumetric loading rate of 11-15 kg COD m(-3).day, 3.6-4.5 h hydraulic retention time and 20 g l(-1) VSS had a scientifically managed technically sound effluent treatment system. While the water quality parameters were found within the acceptable limits, there was 99.07% reduction in BOD, from 43140.0 to 398.0 mg l(-1) and 98.61% reduction in COD from 98003.0 to 1357.0 mg l(-1). There was appreciable improvement in mixed liquor suspended solids (MLSS), mixed liquorvolatile suspended solids (MLVSS) and sludge volume index (SVI). A striking feature was the integrated aerobic-anaerobic highly efficient Up-flow Anaerobic Sludge Blanket (UASB) treatment for biodegradation and energy production that reduced energy and space needs, producing utilisable end-products and net savings on the operational cost. The end-point waste management included terminal products such as fertile sludge, cattle feed supplement, recyclable water and biogas. Vast lagoons with combined aerobic-anaerobic approaches, biogasification unit, sludge recovery, remediated irrigable water were the notable attributes.

Phylogenetic study of methanol oxidizers from chilika-lake sediments using genomic and metagenomic approaches.

Group-wise diversity of sediment methylotrophs of Chilika lake (Lat. 19°28'-19°54'N; Long. 85°06'-85°35'E) Odisha, India at various identified sites was studied. Both the culturable and unculturable (metagenome) methylotrophs were investigated in the lake sediments employing both mxaF and 16S rRNA genes as markers. ARDRA profiling, 16S rRNA gene sequencing, PAGE profiling of HaeIII, EcoRI restricted mxaF gene and the mxaF gene sequences using culture-dependent approach revealed the relatedness of α-proteobacteria and Methylobacterium, Hyphomicrobium and Ancyclobacter sp. The total viable counts of the culturable aerobic methylotrophs were relatively higher in sediments near the sea mouth (S3; Panaspada), also demonstrated relatively high salinity (0.1 M NaCl) tolerance. Metagenomic DNA from the sediments, amplified using GC clamp mxaF primers and resolved through DGGE, revealed the diversity within the unculturable methylotrophic bacterium Methylobacterium organophilum, Ancyclobacter aquaticus, Burkholderiales and Hyphomicrobium sp. Culture-independent analyses revealed that up to 90 % of the methylotrophs were unculturable. The study enhances the general understandings of the metagenomic methylotrophs from such a special ecological niche.

Primary and acquired drug resistance patterns of Mycobacterium tuberculosis isolates in India: a multicenter study.

Tuberculosis is the most prevalent infection worldwide. The emergence of drug-resistant Mycobacterium tuberculosis (M. tuberculosis) isolates emphasizes that it is necessary to monitor drug resistance of the organism against anti-tubercular drugs. We analyzed 327 M. tuberculosis isolates from patients who were cared for at three different health care centers, hereinafter known as study areas (SAs), in North India. Of the 327 total M. tuberculosis isolates, 255 were from a tertiary health care center (Varanasi, Uttar Pradesh [SA-1]), 48 were from a District tuberculosis center (Sawai Madhopur, Rajasthan [SA-2]), and 24 were from a different District tuberculosis center (Buxar, Bihar [SA-3]). Drug susceptibility testing against first-line antibiotics (viz. isoniazid, rifampicin, streptomycin, and ethambutol) was conducted for all the isolates using 1% proportional method. We found that the rates of acquired resistance were consistently higher than the rates of initial drug resistance. In new, untreated cases, a higher degree of MDR-TB was observed at SA-1 (13.3%) and SA-3 (25.0%), whereas it was observed in only 7.1% of the isolates at SA-2. In previously treated patients, MDR cases were found in 35.7% of the isolates from SA-1, 66.6% of the isolates from SA-2, and 43.8% of the isolates from SA-3. Resistance to a single drug was found at a much lower rate, ranging from 0.0 to 6.3% in new cases as well as previously treated cases. In conclusion, the primary resistance of M. tuberculosis is low, but acquired drug resistance is slightly higher in North India.