Characterization and expression profiling of buffalo IFN-lambda family.

Interferon lambda (IFN-λ) is an important type III interferon triggered mainly by viral infection. IFN-λ binds to their heterodimeric receptors and signals through JAK-STAT pathways similar to type I IFN. In this study, we deduced the buffalo IFN-λ sequences through the polymerase chain reaction, and then studied IFN-λ's expression patterns in different tissues, and post induction with poly I:C and live MRSA using RT-qPCR. The full-length sequences of buffalo IFN-λ3, IFN-λ receptors, and a transcript variant of IFN-λ4 were determined. IFN-λ1 is identified as a pseudogene. Virus response elements and a recombination hotspot factor was observed in the regulatory region of IFN-λ. The IFN-λ3 expressed highest in lungs and monocytes but IFN-λ4 did not. The expression of Interferon Lambda Receptor 1 was tissue specific, while Interleukin 10 Receptor subunit beta was ubiquitous. Following poly I:C induction, IFN-λ3 expression was primarily observed in epithelial cells as opposed to fibroblasts, displaying cell type-dependent expression. The cytosolic RNA sensors were expressed highest in endometrial epithelial cells, whereas the endosomal receptor was higher in fibroblasts. 2',5'-oligoadenylate synthetase expressed higher in fibroblasts, myxoma resistance protein 1 and IFN-stimulated gene 56 in epithelial cells, displaying cell-specific antiviral response of the interferon stimulated genes (ISGs). The endometrial epithelial cells expressed IFN-λ3 after live S. aureus infection indicating its importance in bacterial infection. The induction of IFN-λ3 was S. aureus isolate specific at the same multiplicity of infection (MOI). This study elucidates the IFN-λ sequences, diverse expression patterns revealing tissue specificity, and specificity in response to poly I:C and bacterial stimuli, emphasising its crucial role in innate immune response modulation.

Enhanced production of cordycepin in Ophiocordyceps sinensis using growth supplements under submerged conditions.

Cordycepin is a crucial bioactive compound produced by the fungus Cordyceps spp. Its therapeutic potential has been recognized for a wide range of biological properties such as anticancer, anti-diabetic, antidepressant, antioxidant, immunomodulation, etc. Moreover, its human random clinical trials depicted a promising anti-inflammatory activity that reduced the airway inflammation remarkably in asthmatic patients. But its overexploitation and low production of cordycepin in naturally growing biomass are insufficient to meet its existing market demand for its therapeutic use. Therefore, strategies for enhancement of cordycepin production in Cordyceps spp. are warranted. However, specifically, wild type Ophiocordyceps sinensis possesses a very low content of cordycepin and has restricted growth in natural mycelial biomass. To overcome these limitations, this study attempted to enhance cordycepin production in its mycelial biomass in vitro under submerged conditions by adding various growth supplements. The effect of these growth supplements was evaluated by reversed-phase high-performance liquid chromatography (RP-HPLC) which demonstrated that among nucleosides- hypoxanthine and adenosine; amino acids-glycine and glutamine; plant hormones- 1-naphthaleneacetic acid (NAA) and 3-indoleacetic acid (IAA); vitamin-thiamine (B1) from each group of growth supplements yielded a higher amount of cordycepin with 466.48 ±â€¯3.88, 380.23 ±â€¯1.78, 434.97 ±â€¯2.32, 269.78 ±â€¯2.92, 227.61 ±â€¯2.34, 226.02 ±â€¯1.69 and 185.26 ±â€¯2.35 mg/L respectively as compared to control with 13.66 ±â€¯0.64 mg/L. Further, at the transcriptional level, quantitative real time-polymerase chain reaction (qRT-PCR) analysis of genes associated with metabolism and cordycepin biosynthesis depicted significant upregulation of major downstream genes- NT5E, RNR, purA, and ADEK which corroborated well with RP-HPLC analysis. Taken together, the present study identified growth supplements as potential precursors to activate the cordycepin biosynthesis pathway leading to improved cordycepin production in O. sinensis.

Comprehensive Survey on Nanobiomaterials for Bone Tissue Engineering Applications.

One of the most important ideas ever produced by the application of materials science to the medical field is the notion of biomaterials. The nanostructured biomaterials play a crucial role in the development of new treatment strategies including not only the replacement of tissues and organs, but also repair and regeneration. They are designed to interact with damaged or injured tissues to induce regeneration, or as a forest for the production of laboratory tissues, so they must be micro-environmentally sensitive. The existing materials have many limitations, including impaired cell attachment, proliferation, and toxicity. Nanotechnology may open new avenues to bone tissue engineering by forming new assemblies similar in size and shape to the existing hierarchical bone structure. Organic and inorganic nanobiomaterials are increasingly used for bone tissue engineering applications because they may allow to overcome some of the current restrictions entailed by bone regeneration methods. This review covers the applications of different organic and inorganic nanobiomaterials in the field of hard tissue engineering.

Tandem Mass Tag (TMT)-based quantitative proteomics reveals potential targets associated with onset of Sub-clinical Mastitis in cows.

Bovine milk is vital for infant nutrition and is a major component of the human diet. Bovine mastitis is a common inflammatory disease of mammary gland in cattle. It alters the immune profile of the animal and lowers the quality and yield of milk causing huge economic losses to dairy industry. The incidence of sub-clinical mastitis (SCM) is higher (25-65% worldwide) than clinical mastitis (CM) (>5%), and frequently progresses to clinical stage due to lack of sensitive and specific detection method. We used quantitative proteomics to identify changes in milk during sub-clinical mastitis, which may be potential biomarkers for developing rapid, non-invasive, sensitive detection methods. We performed comparative proteome analysis of the bovine milk, collected from the Indian hybrid cow Karan Fries. The differential proteome in the milk of Indian crossbred cows during sub-acute and clinical intramammary gland infection has not been investigated to date. Using high-resolution mass spectrometry-based quantitative proteomics of the bovine whey proteins, we identified a total of 1459 and 1358 proteins in biological replicates, out of which 220 and 157 proteins were differentially expressed between normal and infected samples. A total of 82 proteins were up-regulated and 27 proteins were down-regulated, having fold changes of ≥2 and ≤0.8 respectively. Among these proteins, overexpression of CHI3L1, LBP, GSN, GCLC, C4 and PIGR proteins was positively correlated with the events that elicit host defence system, triggering production of cytokines and inflammatory molecules. The appearance of these potential biomarkers in milk may be used to segregate affected cattle from the normal herd and may support mitigation measures for prevention of SCM and CM.

Fabrication and in-vitro biocompatibility of freeze-dried CTS-nHA and CTS-nBG scaffolds for bone regeneration applications.

The thought of biodegradable organic-inorganic composites composed of natural polymer chitosan and ceramic nanoparticles (hydroxyapatite and bioglass) can be considered as a solution for hard tissue engineering. In this paper, we described a comparative assessment of chitosan-nanohydroxyapatite (CTS-nHA) and chitosan-nano-bioglass (CTS-nBG) scaffolds. The dispersion of nanoscaled hydroxyapatite (nHA) and bioglass (nBG) in chitosan remained satisfactory. The freeze-dried composite based CTS-nHA and CTS-nBG scaffolds shown porous structure. The physiochemical and morphological analysis of all samples has been performed through X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), Brunauer-Emmett-Teller (BET), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). The SEM image confirmed the presence of spherically shaped nHA particles of 4.20 µm and irregularly shaped nBG particles of 6.89 µm. The TEM analysis revealed the existence of 165.52 to 255.17 nm sized nHA particles and 167.35 to 334.69 nm sized nBG particles. TEM analysis also showed the interconnected structure of CTS-nHA and CTS-nBG nanocomposites. After seven days' incubation period, the CTS-nHA and CTS-nBG scaffolds shown good mineralization behavior in simulated body fluid (SBF). The CTS-nHA scaffolds exhibited enhanced compressive strength and elastic modulus compared with the CTS-nBG sample. The cell culture experiment revealed that fabricated scaffolds had good compatibility with fibroblast cells (L929, ATCC) and MG-63 which are able to adhere, proliferate, and migrate through the porous structure. All the obtained results clearly recommend that pre-loaded hydroxyapatite and bioglass nanoparticles can enhance the apatite formation. The scaffolds with chitosan, bioglass, and hydroxyapatite have better biomechanical characteristics and allow cell growth. Therefore, these scaffolds can be perfect candidates for various hard tissue engineering applications such as bone regeneration.

Identification, Optimization of Culture Conditions, and Bioactive Potential of Chinese Caterpillar Mushroom Ophiocordyceps sinensis (Ascomycetes) Mycelium Isolated from Fruiting Body.

The present study deals with the challenges acquainted with in vitro culture of Ophiocordyceps sinensis. We have optimized the culture conditions for the growth of O. sinensis mycelium in semi-synthetic liquid media and determined antibacterial potential of the cultured mycelia extracts. In this study, mycelia were isolated from fruiting bodies and the isolate was identified as O. sinensis anamorph based on sequencing of internal transcribed spacer region. We investigated different culture conditions to optimize the growth of mycelia. Through this investigation, the isolated strain was observed to have its optimum growth at temperature (20°C), which yielded biomass of 12.38 g/L and pH (6.0) yielded biomass of 11.24g/L. Further to augment the production of mycelia, different carbon and nitrogen sources were optimized for mycelium growth in liquid media, out of which sucrose and corn steep powder proved to be the best carbon and nitrogen sources yielding biomass 14.01 g/L and 14.14 g/L, respectively. The evaluation of aqueous and methanolic extracts for antibacterial activity depicted that these extracts are active against all bacterial strains tested here. Aqueous extract depicted minimum inhibitory concentration (MIC) of 0.312, 0.019, 0.078, 0.312, and 0.625 mg/mL and methanolic extract depicted 1.25, 0.078, 0.009, 1.25, and 0.156 mg/mL against Pseudomonas aeruginosa, Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes, respectively. These results led to optimization of enhanced biomass production of O. sinensis, which can be a better alternative approach for further physiological studies and large-scale cultivation of this mushroom for its utilization for therapeutics and nutraceutical values.

Ibuprofen-Loaded CTS/nHA/nBG Scaffolds for the Applications of Hard Tissue Engineering

Background: This study addressed the development of biodegradable and biocompatible scaffolds with enhanced biomechanical characteristics. The biocompatibility and the cationic nature of chitosan (CTS) make it more effective as a bone grafting material. Methods: The hydroxyapatite nanoparticles (nHA) were synthesized by hydrothermal method, and bioglass (nBG) (50% SiO2-45% CaO-5% P2O5) was synthesized using sol-gel method. The ibuprofen-loaded CTS/nHA and CTS/nBG scaffolds were fabricated by using freeze-drying method. Results: Transmission electron microscopy image of nHA and nBG revealed the particles of less than 200 nm. The scanning electron microscopy (SEM) images of CTS/nHA and CTS/nBG scaffolds showed pore sizes ranging from 84-190 µm. The physiochemical characteristics of synthesized ceramic nanoparticles and scaffolds analyzed by XRD were confirmed by ICDD 9-432. The porosity of scaffolds was measured by using SEM, Brunauer-Emmett-Teller method, and Archimedes' principle. The open porosities of CTS/nBG and CTS/nHA samples were 29% and 31%, respectively. The compressive strength of scaffolds was evaluated by stress vs. strain curve. The CTS/nHA scaffold revealed 4% more water retention capacity than CTS/nBG scaffold. In the presence of lysozyme, CTS/nBG scaffold degraded 32.8%, while CTS/nHA degraded 26.1% in PBS solution at pH 7.4. The density of all scaffolds was found (1.9824 g/cm-3 and 1.9338 g/cm-3) to be nearly similar to that of the dry bone (0.8-1.2 g/cm-3). Fibroblast cells multiplied two times in the sample medium of CTS/nBG after 14 days. After 72 h, CTS/nBG and CTS/nHA scaffolds demonstrated 52% and 46% drug release, respectively. Conclusion: Based on our findings, ibuprofen-loaded scaffolds could be an effective drug delivery system for tissue engineering applications.

Efficient protocol for in vitro direct plant regeneration in chickpea Cicer arietinum L.

An efficient plant regeneration system was developed for two important Indian chickpea cultivars, C-235 and HC-1. Immature cotyledons (7-8 mm) directly formed shoots without an intervening callus phase on MS medium containing B5 vitamins, BAP (2.0 mg/l), IBA (0.125 mg/l), AgNO3 (1.69 mg/l) and phytagel (2.5 g/l). The regenerated shoots had normal morphology and were successfully rooted in half strength MS medium under partial dark conditions. Regenerated plants were transferred to potted soil. However, the survival rate of pot house transferred plants was 17.6 per cent.