Kemampuan Whole Cell Helicobacter Pylori Dalam Menginduksi Degradasi Kolagen Tipe IV Melalui Peningkatan Aktivitas Makrofag

Helicobacter pylori (H. pylori) an assosiation with acute myocardial infraction and cronic coronary disease. Bacteri cytotoxin can induce inflamatory of IL1-β and TNFα cytokine produce, and it can acitivated MMP-9 proteolotic enzyme, and it caused collogen degradation in plague atherosclerosis. This research aim to provit the whole cell H.pylori can induce the degradasi of collagen type IV by increase macrophage activity. It were seen from TNFα and IL1-β level use ELISA methode. Anova statistic analyse result show significan defferent (0,000) bettwen group control with macrophage induce whole cell H.pylori. Enzyme MMP-9 production detected with the gelatine zymography continued by western blotting with molecule weight 93 kDa. Collogen fragmentation result of molecule weight which fragment 61,2 kDa and 29 kDa. It concluded that whole cell H.Pylori can induce the degradation of collagen type IV, by increase macrophageag activity. So, the result of this research can support the patomekanism myocardial infract disease

Pengaruh Pemberian Vaksin Whole Cell Killed Virus Terhadap Sintasan Udang Vannamei (Lithopenaeus Vannamei) Yang Diinfeksi Whitespot Baculovirus (WSBV) [the Influence of Whitespot Baculovirus (WSBV) Infection in Lithopenaeus Vannamei by Giving Whole Cell Killed Virus Vaccine on Survival Rate.]

Shrimp are susceptible to a wide variety of pathogens, one of each were viruses. Whitespot Baculovirus (WSBV) is one of virus attack in vannamei. Shrimp had been infected by WSBV will showed high mortality. One of Strategies for prophylaxis and control of WSBV is enhancement of disease resistant by using vaccines. Recently, quasi-immune response have been reported by which Penaeus japonicus surviving from WSBV infections possess a resistance against challenge WSBV (Venegas et al., 2000). This study was used Lithopenaeus vannamei that been vaccination with inactived-formaline WSBV virus cell or it called Whole Cell Killed Virus (WCKV) to know its responses to WSBV. The aim of this study was to know the effect Whole Cell Killed Virus vaccine to survival rate vannamei shrimp (Lithopenaeus vannamei) infected by Whitespot Baculovirus (WSBV). This study used descriptive study. The efficacy of vaccines made of inactivated WSSV with different dose. The dose are P1 (dose 0,01µg/ml each shrimp), P2 (dose 0,02 µg/ml each shrimp), P3 (dose 0,03 µg/ml each shrimp) dan P4 (non-vaccine (kontrol) injected by PBS). Primary parameter was survival rate (%). Secondary parameter was water quality which of temperature, pH, salinity and dissolved oxygen. The result showed that survival rate of L. Vannamei with different dose of WCKV vaccines was increases

Reductive Biotransformation of Ethyl Acetoacetate: A Comparative Studies using Free and Immobilized Whole Yeast Cells

Bioreduction of ethyl acetoacetate with free and immobilized yeast whole cell was achieved by using water and sucrose combination. After detachment from immobilized beads under basic condition, the corresponding ethyl(S)-(+)-3-hydroxybutanoate was isolated with 98 to 100% yield. Immobilized beads of yeast whole cell were prepared at different temperature which affects the morphology and physiology of the beads for the diffusion of the enzyme, which shown the maximum conversion of the substrate to products as compared to the free yeast whole cell

Whole-cell Escherichia coli lactate biosensor for monitoring mammalian cell cultures during biopharmaceutical production

Many high-value added recombinant proteins, such as therapeutic glycoproteins, are produced using mammalian cell cultures. In order to optimise the productivity of these cultures it is important to monitor cellular metabolism, for example the utilisation of nutrients and the accumulation of metabolic waste products. One metabolic waste product of interest is lactic acid (lactate), overaccumulation of which can decrease cellular growth and protein production. Current methods for the detection of lactate are limited in terms of cost, sensitivity, and robustness. Therefore, we developed a whole-cell Escherichia coli lactate biosensor based on the lldPRD operon and successfully used it to monitor lactate concentration in mammalian cell cultures. Using real samples and analytical validation we demonstrate that our biosensor can be used for absolute quantification of metabolites in complex samples with high accuracy, sensitivity and robustness. Importantly, our whole-cell biosensor was able to detect lactate at concentrations more than two orders of magnitude lower than the industry standard method, making it useful for monitoring lactate concentrations in early phase culture. Given the importance of lactate in a variety of both industrial and clinical contexts we anticipate that our whole-cell biosensor can be used to address a range of interesting biological questions. It also serves as a blueprint for how to capitalise on the wealth of genetic operons for metabolite sensing available in Nature for the development of other whole-cell biosensors

The touch and zap method for in vivo whole-cell patch recording of intrinsic and visual responses of cortical neurons and Glial cells

Whole-cell patch recording is an essential tool for quantitatively establishing the biophysics of brain function, particularly in vivo. This method is of particular interest for studying the functional roles of cortical glial cells in the intact brain, which cannot be assessed with extracellular recordings. Nevertheless, a reasonable success rate remains a challenge because of stability, recording duration and electrical quality constraints, particularly for voltage clamp, dynamic clamp or conductance measurements. To address this, we describe "Touch and Zap", an alternative method for whole-cell patch clamp recordings, with the goal of being simpler, quicker and more gentle to brain tissue than previous approaches. Under current clamp mode with a continuous train of hyperpolarizing current pulses, seal formation is initiated immediately upon cell contact, thus the "Touch". By maintaining the current injection, whole-cell access is spontaneously achieved within seconds from the cell-attached configuration by a self-limited membrane electroporation, or "Zap", as seal resistance increases. We present examples of intrinsic and visual responses of neurons and putative glial cells obtained with the revised method from cat and rat cortices in vivo. Recording parameters and biophysical properties obtained with the Touch and Zap method compare favourably with those obtained with the traditional blind patch approach, demonstrating that the revised approach does not compromise the recorded cell. We find that the method is particularly well-suited for whole-cell patch recordings of cortical glial cells in vivo, targeting a wider population of this cell type than the standard method, with better access resistance. Overall, the gentler Touch and Zap method is promising for studying quantitative functional properties in the intact brain with minimal perturbation of the cell's intrinsic properties and local network. Because the Touch and Zap method is performed semi-automatically, this approach is more reproducible and less dependent on experimenter technique

Whole-cell 3D STORM reveals interactions between cellular structures with nanometer-scale resolution.

The ability to directly visualize nanoscopic cellular structures and their spatial relationship in all three dimensions will greatly enhance our understanding of molecular processes in cells. Here we demonstrated multicolor three-dimensional (3D) stochastic optical reconstruction microscopy (STORM) as a tool to quantitatively probe cellular structures and their interactions. To facilitate STORM imaging, we generated photoswitchable probes in several distinct colors by covalently linking a photoswitchable cyanine reporter and an activator molecule to assist bioconjugation. We performed 3D localization in conjunction with focal plane scanning and correction for refractive index mismatch to obtain whole-cell images with a spatial resolution of 20-30 nm and 60-70 nm in the lateral and axial dimensions, respectively. Using this approach, we imaged the entire mitochondrial network in fixed monkey kidney BS-C-1 cells, and studied the spatial relationship between mitochondria and microtubules. The 3D STORM images resolved mitochondrial morphologies as well as mitochondria-microtubule contacts that were obscured in conventional fluorescence images

Whole-cell metabolic control analysis

Since its conception some fifty years ago, metabolic control analysis (MCA) aims to understand how cells control their metabolism by adjusting the activity of their enzymes. Here we extend its scope to a whole-cell context. We consider metabolism in the evolutionary context of growth-rate maximisation by optimisation of protein concentrations. This framework allows for the prediction of flux control coefficients from proteomics data or stoichiometric modelling. Since genes compete for finite biosynthetic resources, we treat all protein concentrations as interdependent. We show that elementary flux modes (EFMs) emerge naturally as the optimal metabolic networks in the whole-cell context and we derive their control properties. In the evolutionary optimum, the number of expressed EFMs is determined by the number of protein-concentration constraints that limit growth rate. We use published glucose-limited chemostat data of S. cerevisiae to illustrate that it uses only two EFMs prior to the onset of fermentation and that it uses four EFMs during fermentation. We discuss published enzyme-titration data to show that S. cerevisiae and E. coli indeed can express proteins at growth-rate maximising concentrations. Accordingly, we extend MCA to elementary flux modes operating at an optimal state. We find that the expression of growth-unassociated proteins changes results from classical metabolic control analysis. Finally, we show how flux control coefficients can be estimated from proteomics and ribosome-profiling data. We analyse published proteomics data of E. coli to provide a whole-cell perspective of the control of metabolic enzymes on growth rate. We hope that this paper stimulates a renewed interest in metabolic control analysis, so that it can serve again the purpose it once had: to identify general principles that emerge from the biochemistry of the cell and are conserved across biological species