Staphylococcal enterotoxin B primes cytokine secretion and lytic activity in response to native bacterial antigens

Superantigens stimulate T-lymphocyte proliferation and cytokine production, but the effects of superantigen exposure on cell function within a complex, highly regulated immune response remain to be determined. In this study, we demonstrate that superantigen exposure significantly alters the murine host response to bacterial antigens in an in vitro coculture system. Two days after exposure to the superantigen staphylococcal enterotoxin B, splenocytes cultured with Streptococcus mutans produced significantly greater amounts of gamma interferon (IFN-gamma) and interleukin-12 than did sham-injected controls. The majority of IFN-gamma production appeared to be CD8(+) T-cell derived since depletion of this cell type dramatically reduced the levels of IFN-gamma. To study host cell damage that may occur following superantigen exposure, we analyzed cytotoxicity to "bystander" fibroblast cells cultured with splenocytes in the presence of bacterial antigens. Prior host exposure to staphylococcal enterotoxin B significantly enhanced fibroblast cytotoxicity in the presence of bacteria. Neutralization of IFN-gamma decreased the amount of cytotoxicity observed. However, a greater reduction was evident when splenocyte-bacterium cocultures were separated from the bystander cell monolayer via a permeable membrane support. Increased cytotoxicity appears to be primarily dependent upon cell-cell contact. Collectively, these data indicate that overproduction of inflammatory cytokines may alter the activity of cytotoxic immune cells. Superantigen exposure exacerbates cytokine production and lytic cell activity when immune cells encounter bacteria in vitro and comparable activities could possibly occur in vivo.     

Calcium can disrupt the SNARE protein complex on sea urchin egg secretory vesicles without irreversibly blocking fusion

The homotypic fusion of sea urchin egg cortical vesicles (CV) is a system in which to correlate the biochemistry and physiology of membrane fusion. Homologues of vesicle-associated membrane protein (VAMP), syntaxin, and SNAP-25 were identified in CV membranes. A VAMP and syntaxin immunoreactive band at a higher apparent molecular mass (approximately 70 kDa) was detected; extraction and analysis confirmed that the band contained VAMP, SNAP-25, and syntaxin. This complex was also identified by immunoprecipitation and by sucrose gradient analysis. VAMP in the complex was insensitive to proteolysis by tetanus toxin. All criteria identify the SNARE complex as that described in other secretory systems. Complexes exist pre-formed on individual CV membranes and form between contacting CV. Most notably, CV SNARE complexes are disrupted in response to [Ca2+]free that trigger maximal fusion. N-Ethylmaleimide, which blocks fusion at or before the Ca2+-triggering step, blocks complex disruption by Ca2+. However, disruption is not blocked by lysophosphatidylcholine, which transiently arrests a late stage of fusion. Since removal of lysophosphatidylcholine from Ca2+-treated CV is known to allow fusion, complex disruption occurs independently from the membrane fusion step. As Ca2+ disrupts rather than stabilizes the complex, the presumably coiled-coil SNARE interactions are not needed at the time of fusion. These findings rule out models of fusion in which SNARE complex formation goes to completion ("zippers-up") after Ca2+ binding removes a "fusion-clamp."     

Traffic of dynamin within individual Drosophila synaptic boutons relative to compartment-specific markers

Presynaptic terminals contain several specialized compartments, which have been described by electron microscopy. We show in an identified Drosophila neuromuscular synapse that several of these compartments-synaptic vesicle clusters, presynaptic plasma membrane, presynaptic cytosol, and axonal cytoskeleton-labeled by specific reagents may be resolved from one another by laser scanning confocal microscopy. Using a panel of compartment-specific markers and Drosophila shibire(ts1) mutants to trap an intermediate stage in synaptic vesicle recycling, we have examined the localization and redistribution of dynamin within single synaptic varicosities at the larval neuromuscular junction. Our results suggest that dynamin is not a freely diffusible molecule in resting nerve terminals; rather, it appears localized to synaptic sites by association with yet uncharacterized presynaptic components. In shi(ts1) nerve terminals depleted of synaptic vesicles, dynamin is quantitatively redistributed to the plasma membrane. It is not, however, distributed uniformly over presynaptic plasmalemma; instead, fluorescence images show "hot spots" of dynamin on the plasma membrane of vesicle-depleted nerve terminals. We suggest that these dynamin-rich domains may mark the active zones for synaptic vesicle endocytosis first described at the frog neuromuscular junction.     

Robust kernelized approach to clustering by incorporating new distance measure

A new data clustering algorithm Density oriented Kernelized version of Fuzzy c-means with new distance metric (DKFCM-new) is proposed. It creates noiseless clusters by identifying and assigning noise points into separate cluster. In an earlier work, Density Based Fuzzy C-Means (DOFCM) algorithm with Euclidean distance metric was proposed which only considered the distance between cluster centroid and data points. In this paper, we tried to improve the performance of DOFCM by incorporating a new distance measure that has also considered the distance variation within a cluster to regularize the distance between a data point and the cluster centroid. This paper presents the kernel version of the method. Experiments are done using two-dimensional synthetic data-sets, standard data-sets referred from previous papers like DUNN data-set, Bensaid data-set and real life high dimensional data-sets like Wisconsin Breast cancer data, Iris data. Proposed method is compared with other kernel methods, various noise resistant methods like PCM, PFCM, CFCM, NC and credal partition based clustering methods like ECM, RECM, CECM. Results shown that proposed algorithm significantly outperforms its earlier version and other competitive algorithms.     

Hardystonite improves biocompatibility and strength of electrospun polycaprolactone nanofibers over hydroxyapatite: A comparative study

The aim of this study was to compare physico-chemical and biological properties of hydroxyapatite (HA) and hardystonite (HS) based composite scaffolds. Hardystonite (Ca₂ZnSi₂O₇) powders were synthesized by a sol–gel method while polycaprolactone–hardystonite (PCL–HS) and polycaprolactone–hydroxyapatite (PCL–HA) were fabricated in nanofibrous form by electrospinning. The physico-chemical and biological properties such as tensile strength, cell proliferation, cell infiltration and alkaline phosphatase activity were determined on both kinds of scaffolds. We found that PCL–HS scaffolds had better mechanical strength compared to PCL–HA scaffolds. Addition of HA and HS particles to PCL did not show any inhibitory effect on blood biocompatibility of scaffolds when assessed by hemolysis assay. The in vitro cellular behavior was evaluated by growing murine adipose-tissue-derived stem cells (mE-ASCs) over the scaffolds. Enhanced cell proliferation and improved cellular infiltrations on PCL–HS scaffolds were observed when compared to HA containing scaffolds. PCL–HS scaffolds exhibited a significant increase in alkaline phosphatase (ALP) activity and better mineralization of the matrix in comparison to PCL–HA scaffolds. These results clearly demonstrate the stimulatory role of Zn and Si present in HS based composite scaffolds, suggesting their potential application for bone tissue engineering.     

Vapor Phase Selective Catalytic Oxidation of Cyclohexane over 13X Supported Metal Oxides in the Presence of Ozone

This article reports the application of ozone for the selective oxidation of cyclohexane over 13X molecular sieve supported various metal oxides at ambient temperatures. From the SEM, XRD and HR-TEM results, the impregnated metal oxides are highly dispersed on the support. The activity results reveal that Co/MS, Mo/MS, Cu/MS, and Ag/MS catalysts produce cyclohexanone/cyclohexanol as selective oxidation products, whereas Ce/MS, Mn/MS, and V/MS catalysts yield, predominantly, CO and CO₂. Among them, Co/MS catalyst exhibits better conversion of 12.2% with selectively of 58% to cyclohexanone/cyclohexanol, which is attributed to the simultaneous activation of ozone and cyclohexane (-C-H bond) at ambient conditions.     

Unique Physico-chemical Properties of Sal (Shorea robusta) Starch

Starch from sal seeds cake was isolated, bleached and purified. Scanning electron microscopy of the starch showed round to oval shaped granules of the size 3.5 to 12.3 μm. The starch has more nitrogen, phosphorus, lipids, amylose and water binding capacity in comparison to the other starches of forest origin. Despite its low and seemingly restricted swelling, the sal starch is much more soluble at any particular degree of swelling. Paste viscosity curve showed no drop in viscosity after full gelatinisation had taken place, i. e. the paste is stable on continued cooking.     

Evaluation of antimicrobial effects of lauric arginate on reduction of Salmonella spp. in ground chicken

The efficacy of lauric arginate (LAE) on Salmonella survivability in ground chicken containing 9.8% fat was determined under refrigerated storage. The effect of LAE treatments on total aerobes, pH and objective colour were also assessed in ground chicken. LAE was highly effective against Salmonella in 0.1% peptone with no detectable survivors following treatment with 200 ppm and 400 ppm of LAE. No difference existed (P > 0.05) in the growth of Salmonella and total aerobes after treatment with 200 and 400 ppm of LAE in ground chicken inoculated with a four strain mixture of Salmonella. At these concentrations, LAE did not exhibit any treatment effect on the pH and colour of ground chicken (P > 0.05). In conclusion, although LAE possesses strong inhibitory (P < 0.05) effect against Salmonella in suspension in 0.1% peptone water, no inhibitory (P > 0.05) effect on growth of Salmonella was observed in ground chicken at the currently approved levels of 200 ppm of LAE.