Biosynthesis of xylobiose: a strategic way to enrich the value of oil palm empty fruit bunch fiber

Xylooligosaccharides are functional foods mainly produced during the hydrolysis of xylan by physical, chemical, or enzymatic methods. In this study, production of xylobiose was investigated using oil palm empty fruit bunch fiber (OPEFB) as a source material, by chemical and enzymatic methods. Xylanase-specific xylan hydrolysis followed by xylobiose production was observed. Among different xylanases, xylanase from FXY-1 released maximum xylobiose from pretreated OPEFB fiber, and this fungal strain was identified as Aspergillus terreus and subsequently deposited under the accession Number MTCC- 8661. The imperative role of lignin on xylooligosaccharides enzymatic synthesis was exemplified with the notice of xylobiose production only with delignified material. A maximum 262 mg of xylobiose was produced from 1.0 g of pretreated OPEFB fiber using FXY-1 xylanase (6,200 U/ml) at pH 6.0 and 45° C. At optimized environment, the yield of xylobiose was improved to 78.67 g/100 g (based on xylan in the pretreated OPEFB fiber).     

Interaction of anesthetic supplement thiopental with human serum albumin

Thiopental (TPL) is a commonly used barbiturate anesthetic. Its binding with human serum albumin (HSA) was studied to explore the anesthetic-induced protein dysfunction. The basic binding interaction was studied by UV-absorption and fluorescence spectroscopy. An increase in the binding affinity (K) and in the number of binding sites (n) with the increasing albumin concentration was observed. The interaction was conformation-dependent and the highest for the F isomer of HSA, which implicates its slow elimination. The mode of binding was characterized using various thermodynamic parameters. Domain II of HSA was found to possess a high affinity binding site for TPL. The effect of micro-metal ions on the binding affinity was also investigated. The molecular distance, r, between donor (HSA) and acceptor (TPL) was estimated by fluorescence resonance energy transfer (FRET). Correlation between the stability of the TPL-N and TPL-F complexes and drug distribution is discussed. The structural changes in the protein investigated by circular dichroism (CD) and Fourier transform infrared (FT-IR) spectroscopy reflect perturbation of the albumin molecule and provide an explanation for the heterogeneity of action of this anesthetic.     

Peroxidase and catalase changes during in vitro adventitious shoot organogenesis from hypocotyls of Albizia odoratissima L.f. (Benth)

Hypocotyls of Albizia odoratissima cultured on shoot induction medium (MS medium with 7.5 μM BAP and 0.5 μM NAA) showed adventitious shoot organogenesis under light with 16 h photoperiod. Similar cultures under total darkness produced non-morphogenic calli. The changes in the specific peroxidase and catalase activity, total protein content and acidic isoperoxidase pattern were compared between the culture showing shoot organogenesis and culture producing non-morphogenic calli. It was found that in vitro shoot bud differentiation is accompanied by an increase of the specific activities of peroxidase and catalase in culture kept under light. In parallel with the above changes the total protein content reached to the maximum level and also a new isoperoxidase (P10) expressed on the 21st day in cultures kept under light. Conversely, culture producing non-morphogenic calli underwent a reverse change in specific peroxidase activity. This change in antioxidant enzyme activities corresponds to the histological observation of shoot bud differentiation in cultures kept under light.     

Attenuation of dengue virus infection by adeno-associated virus-mediated siRNA delivery

BACKGROUND: The need for safe and effective treatment of dengue virus (DEN), a class A agent that causes dengue hemorrhagic fever/dengue shock syndrome, has been a critical global priority. An effective vaccine for DEN is not yet available. In this study the possibility of attenuating DEN infection using adeno-associated virus (AAV)-encoded short interfering RNAs (siRNA) was examined in Vero cells and human dendritic cells (DCs). METHODS: A cassette encoding siRNA targeted to a 3' untranslated sequence common to all DEN serotypes was designed and tested for its ability to attenuate DEN infection by use of AAV delivery. RESULTS: Vero cells or DCs infected with AAV-siRNA showed a significant, dose-dependent reduction in DEN infection. Treatment of DCs with AAV-siRNA also decreased the DEN-induced apoptosis of DCs and did not induce significant inflammation. CONCLUSION: These results demonstrate that AAV-mediated siRNA delivery is capable of reducing DEN infection in cells and may be useful in decreasing DEN replication in humans.     

SWAP-70 regulates c-kit-induced mast cell activation, cell-cell adhesion, and migration

SWAP-70, an unusual phosphatidylinositol-3-kinase-dependent protein that interacts with the RhoGTPase Rac, is highly expressed in mast cells. Cultured bone marrow mast cells (BMMC) from SWAP-70(-/-) mice are reduced in FcepsilonRI-triggered degranulation. This report describes the hitherto-unknown role of SWAP-70 in c-kit receptor signaling, a key proliferation and differentiation pathway in mast cells. Consistent with the role of Rac in cell motility and regulation of the actin cytoskeleton, mutant cells show abnormal actin rearrangements and are deficient in migration in vitro and in vivo. SWAP-70(-/-) BMMC are impaired in calcium flux, in proper translocation and activity of Akt kinase (required for mast cell activation and survival), and in translocation of Rac1 and Rac2 upon c-kit stimulation. Adhesion to fibronectin is reduced, but homotypic cell association induced through c-kit is strongly increased in SWAP-70(-/-) BMMC. Homotypic association requires extracellular Ca(2+) and depends on the integrin alpha(L)beta(2) (LFA-1). ERK is hyperactivated upon c-kit signaling in adherent and dispersed mutant cells. Together, we suggest that SWAP-70 is an important regulator of specific effector pathways in c-kit signaling, including mast cell activation, migration, and cell adhesion.     

Selection intensities and consanguineity in the Yadava and Vadabalija of Visakhapatnam, Andhra Pradesh, India.

The opportunity for intensity of selection is studied in two backward caste groups with different occupations, namely Yadava (pastoral) and Vadabalija (fishing) of Bheemunipatnam Taluk of Visakhapatnam District, Andhra Pradesh. The amount of selection intensity is found to be higher in Vadabalija (0.8583) than Yadava (0.5827). The contribution of mortality to the total index of selection is greater in Vadabalija (0.6095) than in Yadava (0.3326). When consanguineity is considered, the fertility components of selection intensity (0.1885) are lower in the consanguineous group of Yadava than in the nonconsanguineous group (0.2845), whereas in Vadabalija they are higher in the consanguineous group (0.5261) than in the nonconsanguineous group (0.2223).     

Advances in polymeric systems for tissue engineering and biomedical applications

The characteristics of tissue engineered scaffolds are major concerns in the quest to fabricate ideal scaffolds for tissue engineering applications. The polymer scaffolds employed for tissue engineering applications should possess multifunctional properties such as biocompatibility, biodegradability and favorable mechanical properties as it comes in direct contact with the body fluids in vivo. Additionally, the polymer system should also possess biomimetic architecture and should support stem cell adhesion, proliferation and differentiation. As the progress in polymer technology continues, polymeric biomaterials have taken characteristics more closely related to that desired for tissue engineering and clinical needs. Stimuli responsive polymers also termed as smart biomaterials respond to stimuli such as pH, temperature, enzyme, antigen, glucose and electrical stimuli that are inherently present in living systems. This review highlights the exciting advancements in these polymeric systems that relate to biological and tissue engineering applications. Additionally, several aspects of technology namely scaffold fabrication methods and surface modifications to confer biological functionality to the polymers have also been discussed. The ultimate objective is to emphasize on these underutilized adaptive behaviors of the polymers so that novel applications and new generations of smart polymeric materials can be realized for biomedical and tissue engineering applications.     

Effect of legume-based cropping systems on nitrogen mineralization potential of Vertisol

The quantity and patterns of net mineralization of soil nitrogen (N) were studied in Vertisols under different cropping systems in the semi-arid tropical areas. Eight cropping systems were selected; three contained pigeonpea (PP), one contained PP and cowpea (COP), and two contained chickpea (CP) as legume component crops, one included sequence cropping with nonlegumes during the rainy and postrainy seasons, and one system was kept fallow (F) during the rainy season and sown to sorghum (S) during the postrainy season. Cropping systems with PP as a component crop increased mineralizable N(N _o ) content two-fold in the soil compared with fallow + sorghum (F+S)–F+S system. The N mineralization rate constant (k) was not significantly affected by previous cropping history of the soil; however, a numerically higher rate constant was observed in the COP/PP intercrop, followed by sequential S+safflower (SF) system as compared to the other soils. Mineral N accumulation curves for six soils were more accurately described by the exponential model than the linear model. The active N fraction (N _o /N_tot %) varied between 8 and 16% for different systems and a direct relationship was observed between N _o /N_tot and total N for the soils under diverse cropping systems.ICRISAT JA (1638)     

Purification and characterization of flavokinase from Neurospora crassa

The ATP-dependent phosphorylation of riboflavin to FMN by flavokinase is the key step in flavin biosynthesis. Flavokinase has been purified from a fungal source for the first time. The enzyme purified from a cell wall lacking mutant of Neurospora crassa, slime, is a monomer of M(r) 35.5 kDa with maximal activity at alkaline pH and high temperature (55 degrees C). The K(m) for both substrates is the lowest reported for flavokinase from any source so far (120 nM for riboflavin and 210 nM for MgATP2-). The enzyme exhibits preference for Mg2+ over Zn2+ as the essential activator and is also significantly activated by several cations. Activation by orthophosphate may be physiologically relevant for the intracellular regulation of flavokinase.     

The significance of alpha 5 beta 1 integrin dependent and independent actin cytoskelton organization in cell transformation and survival

Cell-matrix and cell-cell interactions are important physiological determinants of cell growth, survival and transformation. Cell adhesion to the extra cellular matrix (ECM) via integrins also crucially influences the organization of the cytoskeleton. It triggers a cascade of intracellular biochemical events, which regulate cell viability and growth. We have studied the relationship between cell attachment to the substratum and cytoskeletal organization and cell survival and transformation. Our results demonstrate that in the absence of attachment to the substratum, adhesion-dependent fibroblasts exhibit rapid loss of viability. However, a small percentage of cells survive even after remaining non-adherent for 16h. The adherent and non-adherent cells differ from one another both morphologically and physiologically. The latter show a loss of alpha5beta1 integrin expression on their surface and bind non-specifically to the substratum and ECM, thereby activating certain pathways more efficiently than adherent cells. We have also shown that non-adherent cells grow faster and have worse cytoskeletal organization after attachment to the substratum, and do not form focal adhesions or actin stress fibres. Hence, our data suggests that rat fibroblasts in prolonged suspension exhibit some properties that are comparable to cells undergoing transformation, by adapting integrin-dependent or independent signalling pathways for their survival.