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Aims Numerous pre-clinical and clinical studies have recently demonstrated the significant role of phage therapy in treating multidrug-resistant bacterial infections. However, only a few researchers have focused on monitoring the phage-mediated adverse reactions during phage therapy. The present study aimed to demonstrated the oral acute and sub-acute toxicity of bacteriophages (Klebsiella pneumoniae XDR strain) in Charles Foster rats with special reference to immunological response and adverse effects. Methods and Results Bacteriophages were orally administered in dosages of 1010 PFU/ml and a 1015 PFU/ml to Charles Foster rats as a single dose (in acute toxicity study) and daily dosage for 28 days (in sub-acute toxicity study). One millilitre suspension of bacteriophages was administered through the oral gavage feeding tube. No adverse effect was observed in any of the experimental as well as in the control animals. Furthermore, an insignificant change in food and water intake and body weight was observed throughout the study period compared with the control group rats. On the 28th day of phage administration, blood was collected to estimate haematological, biochemical and cytokines parameters. The data suggested no difference in the haematological, biochemical and cytokine profiles compared to the control group. No significant change in any of the treatment groups could be observed on the gross and histopathological examinations. The cytokines estimated, interleukin-1 beta (IL-1β), IL-4, IL-6 and IFN-gamma, were found within the normal range during the experiment. Conclusions The results concluded that no adverse effect, including the severe detrimental impact on oral administration of high (1010 PFU/ml) and very high dose (1015 PFU/ml) of the bacteriophages cocktail. Significance and Impact of Study The high and long-term oral administration of bacteriophages did not induce noticeable immunological response as well.

Background: Scrub typhus (ST) is a rickettsial infection caused by Orientia tsutsugamushi, which presents with flu like symptoms. This disease has been reported from all over India but with slight variations in its pattern. For decreasing the prevalence, preventing new incidences, and predicting the course of the ST, therefore, it is crucial to gain knowledge and perception of local risk components associated with the disease. The present study aimed to investigate the epidemiological distribution and potential risk factors of O. tsutsugamushi Infection in Eastern Uttar Pradesh (EUP), India. Methods: The serums of 211 samples were collected from the suspected cases along with the detailed information about the participants such as age, location, and place recorded in case history form (CRF). IgM estimation was performed using enzyme-linked immunosorbent assay (ELISA) assay. Results: A total of 58 samples (27.4%) out of 211 ones were found to be positive for IgM antibodies against O. tsutsugamushi bacterium. Furthermore, the results were correlated with epidemiological data such as gender, rural or urban background, pets, and occupation. The results showed that 76.7% of the study participants were from rural areas or had bushes around their houses, 88.3% of them had pets/cattle or frequent encounter with rodents at their houses, and 30.3% of them had no toilet facilities at home. Conclusions: It was concluded that the proximity to pets/cattle, having rodents in closer vicinity, residing in places surrounded by vegetation/farm/bushy areas, and following occupations involving field work increased the chances of getting bitten by mites/chiggers. Overall, Orientia tsutsugamushi prevalence increased in EUP, with respect to clinical features, disease presentation, and laboratory diagnosis can help our community to reduce the mortality caused by this infectious disease.

The study is aimed at establishing the optimal parameters for RNA purification of pooled specimens, in SARS-CoV-2 assay. This research work evaluates the difference of extracted RNA purity of pooled samples with and without treatment with isopropyl alcohol and its effect on real-time RT-PCR. As per the protocol of the Indian Council of Medical Research (ICMR), 5 sample pools were analysed using qRT-PCR. A total of 100 pooled samples were selected for the study by mixing 50 μL of one COVID-19 positive nasopharyngeal/oropharyngeal (NP/OP) specimen and 50 μL each of 4 known negative specimens. Pool RNA was extracted using the column-based method, and 1 set of pooled extracted RNA was tested as such, while RNA of the second set was treated additionally with chilled isopropyl alcohol (modified protocol). Further, the purity of extracted RNA in both the groups was checked using Microvolume Spectrophotometers (Nanodrop) followed by RT-PCR targeting E-gene and RNaseP target. The results showed that the purity index of extracted RNA of untreated pooled specimens was inferior to isopropyl alcohol-treated templates, which was observed to be 85% sensitivity and 100% specificity. The average Cq (E gene) in the unpurified and purified pool RNA group was 34.66 and 31.48, respectively. The nanodrop data suggested that purified RNA concentration was significantly increased with an average value of 24.73 ± 1.49 ng / uL , which might be the reason for high sensitivity and specificity. Thus, this group testing of SARS-CoV-2 cases using pools of 5 individual samples would be the best alternative for saving molecular reagents, personnel time, and can increase the overall testing capacity. However, purity of RNA is one of the important determinants to procure unfailing results, thus, this additional purification step must be included in the protocol after RNA has been extracted using commercially available kit before performing qRT-PCR.

Significant genetic diversity was observed in 218 out of a total of 1309 accessions of amaranth (Amaranthus hypochondriacus L.) and its seven wild relatives, A. spinosus L., A. dubius Mart. ex Thell., A. hybridus L., A. tricolor L., A. cruentus L., A. caudatus L., A. retroflexus L. for 24 nutritional parameters including total oil content, fatty acid profile, total protein content and amino acid profile. Diversity for total oil content (6.42–12.53%), linoleic acid (25.68–54.34%), oleic acid (21.97–42.01%) of the total fatty acids, total protein content (7.84–18.01%), among important essential amino acids; lysine content (0.66–11.12 g/16 g N), methionine (0.35–4.80 g/16 g N) and half cystine and (0.12–8.32 g/16 g N) was reported. The un-weighted pair-group method using arithmetic average cluster analysis based on pair wise Euclidean genetic distance grouped the accessions into seven major clusters. Histidine, half cystine, tyrosine, essential amino acids, total oil content, linoleic acid and oleic acid content were the major parameters contributing significantly to genetic diversity. Present findings indicate that significant diversity exists for nutritional parameters in amaranth germplasm. The promising accessions with higher multiple nutritive traits; protein content (>16%), oil content (>11%), lysine content (>7.5 g/16 g N) and EAA higher than the FAO reported values, were identified. This is the first report on detailed nutritional analysis of diversity collected worldwide. These could be used as potential breeding material for nutritional enhancement through genetic improvement. This will help in overcoming the “triple burden” of malnourishment, hidden hunger, and obesity.

Pancreatic acinar cells secrete fluid and digestive enzymes. Both types of secretion are activated by a rise in intracellular calcium but how the stimulus-secretion cascade actually regulates secretory output is not well understood. It has long been known that the calcium response of acinar cells to physiological stimulation is complex. Dependent on the type and concentration of agonist, it consists of either local or global calcium increases as well as spreading waves of calcium across the cell. In the past it has been speculated that these different calcium signals drive different secretory responses. Now, recent employment of two-photon microscopy has enabled the simultaneous recording of both enzyme secretion and calcium signals and is beginning to resolve this issue. The data shows that local calcium responses exclusively drive fluid secretion. Where-as, global calcium responses drive both fluid and enzyme secretion. This differential control of secretory output is likely central to controlling the physiological responses of pancreatic acinar cells.

1. Herein we review past work that has studied the composition of luminal fluid in organs, with a focus on measures of calcium and pH in the exocrine glands. This luminal environment is 'external' to the mammalian body and is not subject to the usual mechanisms of homeostatic control. Instead, it is controlled by the behaviour of the cells that line the lumen. 2. We discuss the likely possibility that rapid and local changes in calcium and pH occur within microdomains in the lumen. Further, we present preliminary evidence, using live cell imaging of intact pancreatic fragments, that supports the idea that pH changes do occur. Our evidence indicates that exocytosis of secretory granules in pancreatic acinar cells leads to a loss of protons from the granule and a subsequent local acidification of the lumen. 3. These changes in luminal composition are placed in the context of diseases of the pancreas, such as cystic fibrosis and pancreatitis, both of which are known to result in perturbations of luminal fluid composition.

Wireless ethernet is one of the most important wireless technology that enhances our personal environment, either work or private, by means of networking or a variety of personal and wearable devices within the space and with the outside world. It refers to a link layer protocol in the TCP/IP (Transmission Control Protocol/Internet Protocol) stack, describing how networked devices can format data for transmission to other network devices on the same network segment, and how to put that data out on the network connection. This paper represents an overall view of the wireless Ethernet features, advantages, applications and integration of mobile communication. It initially assumed a shared medium: multiple devices on each segment of the network, connected daisy chain at first but later in star topology via Ethernethubs(which replicated all traffic received on any port to every other port).

The diflavo-protein NADPH cytochrome P450 reductase (CPR) is the key electron transfer partner for all drug metabolizing cytochrome P450 enzymes in humans. The protein delivers, consecutively, two electrons to the heme active site of the P450 in a carefully orchestrated process which ultimately leads to the generation of a high valent oxo-heme moiety. Despite its central role in P450 function, no direct electrochemical investigation of the purified protein has been reported. Here we report the first voltammetric study of purified human CPR where responses from both the FMN and FAD cofactors have been identified using both cyclic and square wave voltammetry. For human CPR redox responses at −2 and −278 mV (with a ratio of 1e−:3e−) vs NHE were seen at pH 7.9 while the potentials for rat CPR at pH 8.0 were −20 and −254 mV. All redox responses exhibit a pH dependence of approximately −59 mV/pH unit consistent with proton coupled electron transfer reactions of equal stoichiometry. Keywords: Cytochrome P450 reductase, Protein, Voltammetry

The human cytochromes P450 are responsible for the clearance of ∼90% of xenobiotics yet comparatively little is known about their electrochemistry. Here we report the first direct electrochemistry of P450s from the 2C subfamily; one of the major groups of enzymes from this family. Specifically, the proteins that we have examined are recombinant human P450s 2C9, 2C18 and 2C19 and reversible FeIII/II couples are seen in the absence of dioxygen. Even in the presence of trace amounts of dioxygen, a pronounced cathodic response is seen which is assigned to catalytic reduction of the bound dioxygen ligand by the ferrous P450. Keywords: Cytochrome P450, Voltammetry, Direct electron transfer

This paper presents the data for the effect of adsorbent dose, initial sorbate concentration, contact time, and pH on the adsorption of chromium(VI) on maple sawdust. Batch adsorption studies have been carried out. An empirical relationship has been obtained to predict the percentage chromium(VI) removal at any time for known values of sorbent and initial sorbate concentration. Under observed test conditions, the equilibrium adsorption data fits the linear Langmuir and Freundlich isotherms. The experimental result inferred that chelation ion exchange is one of the major adsorption mechanisms for binding metal ions to the maple sawdust.

In this work, sorption of lead on sawdust (SD) has been studied by using batch techniques. Similar to the sorption of copper, the equilibrium sorption levels for lead is a function of the solution pH, contact time, sorbent and sorbate concentration. The equilibrium adsorption capacity of sawdust for lead was measured and extrapolated using linear Freundlich and Langmuir isotherms and compared with that for copper. Metal ions which are bounded to the sawdust could be stripped by acidic solution so that the sawdust can be recycled. Non-equilibrium experiments were done by using glass columns. The results indicate the potentially practical value of this method in industries and also provide strong evidence to support the adsorption mechanism proposed. Even though the focus of this work is on lead, some results from previous study for copper have been incorporated here for sake of comparison.

Locally available sawdust, a byproduct of the world industry, was found to be a low cost and promising adsorbent for the removal of copper(II) from wastewater. In this work, adsorption of copper on sawdust has been studied by using batch techniques. The equilibrium adsorption level was determined to be a function of the solution pH, contact time, and sorbate concentration. The equilibrium adsorption capacity of sawdust for copper was obtained by using linear Freundlich and Langmuir isotherms. Ion exchange is probably one of the major adsorption mechanisms for binding divalent metal ions to the maple sawdust. The results provide strong evidence to support the hypothesis of adsorption mechanism. It also supplies significant sawdust operation capacity data for engineering design.

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific potent mitogen involved in a number of pathologic processes, including angiogenesis, tumor growth, and metastasis. Polymorphisms of VEGF gene have been associated with susceptibility to several cancers. VEGF gene susceptibility to bladder cancer (BC), however, still remains controversial. We analyzed association of the -2578C/A, -7C/T, -2549I/D, and -1001G/C polymorphism of the VEGF gene with bladder cancer (BC) risk and interaction of these polymorphisms with lifestyle and demographic factors. We genotyped -2578C/A, -7C/T, -2549I/D and -1001G/C by PCR-restriction fragment length polymorphism and amplification refractory mutation specific method to evaluate risk in 200 BC patients and 250 healthy controls from North Indian population. Significant association for BC risk in heterozygous CA genotype (1.69-fold) in VEGF-2578C/A and heterozygous genotype of VEGF-1001G/C (p = 0.032) was observed. Interestingly, VEGF-2549I/D genotype showed reduced risk for BC. The gene-gene combination analysis revealed DD-GG with reduced risk (p = 0.018) of VEGF-2549I/D and VEGF-1001G/C, and combination CA-GG of VEGF-2578C/A and VEGF-1001G/C demonstrated 1.75-fold risk for BC. Our findings suggested that polymorphism -2578C/A and -1001G/C in the promoter of VEGF gene may play a significant role in mediating the bladder cancer risk, whereas VEGF-2549I/D genotype appears to be protective for BC.

The relationship between the cellular Ca 2+ signal and secretory vesicle fusion (exocytosis) is a key determinant of the regulation of the kinetics and magnitude of the secretory response. Here, we have investigated secretion in cells where the exocytic response is controlled by Ca 2+ release from intracellular Ca 2+ stores. Using live-cell two-photon microscopy that simultaneously records Ca 2+ signals and exocytic responses, we provide evidence that secretion is controlled by changes in Ca 2+ concentration [Ca 2+ ] in relatively large-volume microdomains. Our evidence includes: (1) long latencies (>2 seconds) between the rise in [Ca 2+ ] and exocytosis, (2) observation of exocytosis all along the lumen and not clustered around Ca 2+ release hot-spots, (3) high affinity ( K d =1.75 μM) Ca 2+ dependence of exocytosis, (4) significant reduction in exocytosis in the prescence of cytosolic EGTA, (5) spatial exclusion of secretory granules from the cell membrane by the endoplasmic reticulum, and (6) inability of local Ca 2+ responses to trigger exocytosis. These results strongly indicate that the control of exocytosis, triggered by Ca 2+ release from stores, is through the regulation of cytosolic [Ca 2+ ] within a microdomain.

The chlorosulfides 1 or 2 react with silyl enol ethers in presence of anhydrous zinc bromide to give mainly the corresponding syn products 4 or 6 respectively. The high syn selectivity of these reaction is explained by nucleophilic addition to a Chelated Chiral thioniun ion.

Solidification/stabilization of toxic pentachlorophenol (PCP) has been studied by using spectroscopic and electrochemical techniques. Portland type I c

Amino terminal sequence modification of cytochrome P450 enzymes is often necessary to achieve expression in bacteria. The aim of this study was to examine the effect of such modifications on membrane integration and P450 activity. Forms that retained substantial N-terminal hydrophobic sequences remained unaffected by treatments to remove peripheral membrane proteins and were released only by detergent. Truncated P450s 2A13, 2C9 (delta 3-20), 2C19 (delta 3-20), 2D6 (DB11) and 2E1 remained principally membrane-bound, but some P450 was found in the soluble fraction and could be partially extracted by alkaline and high salt treatments. The subcellular localization of P450s 2C9 and 2C19 assessed by fluorescence microscopy mirrored the distribution between subcellular fractions. The MALLLAVFL modified forms of P450 2C9 YFP, P450 2C18 YFP and P450 2C19 YFP were found primarily at the periphery of the cells, whereas the truncated forms of P450 2C9 (delta 3-20) YFP and 2C19 (delta 3-20) YFP were observed at the periphery as well as inside the cells. N-terminal variants of P450s 2C9 and 2C19 showed altered kinetics towards form-selective substrates. Rates of diclofenac 4 -hydroxylation by P450 2C9 and luciferin H-EGE metabolism by P450 2C19 were higher for the MALLLAVFL-modified forms compared with the (delta 3-20) truncated forms despite supplementation of truncated form incubations with additional reductase. Thus, N-terminal sequence modifications changed the degree of membrane integration, potentially affecting subcellular localization, interactions with redox partners, and hence enzymatic activity.

The chlorosulfides 1 or 2 react with silyl enol ethers in presence of anhydrous zinc bromide to give mainly the corresponding syn products 4 or 6 respectively. The high syn selectivity of these reaction is explained by nucleophilic addition to a Chelated Chiral thioniun ion.

Oxiranes are regioselectively cleaved by benzenethiol in presence of cobalt(II) chloride or Co2(CO)8 to the corresponding β-hydroxy sulfides in good yields. A mechanistic study of this reaction reveals that an electron transfer process is involved in these transformations and the reaction is proceeding via the formation of a carbon-cobalt bond.

The authors' objective in this study was to introduce and evaluate integrated cell culture polymerase chain reaction (ICC-PCR) as a technique for the rapid screening of poliovirus in sewage samples. Researchers are in the last stage of poliomyelitis eradication; however, in a densely populated country such as India, time is the prime factor in the identification of poliovirus circulation and transmission because this virus follows the fecal-oral route for transmission and is excreted in nature. The authors used ICC-PCR to detect poliovirus in sewage samples, and they compared this nonconventional method with conventional cell culture methods to determine sensitivity, accuracy, and the time from sample collection to final results. The ICC-PCR method provided results within 4-5 days of sewage-sample collection; in contrast, the conventional method takes more than 18 days to provide such results. The ICC-PCR method proved to be sensitive, reproducible, and accurate, as well as rapid in its screening of sewage samples for poliovirus. This diagnostic tool may indeed prove quite useful in polio eradication.

The main parameters influencing Nickel (II) metal sorption on maple sawdust were: initial metal ion concentration, amount of adsorbent, and pH value of solution. The maximum percent metal removal was attained after about 1 h. The greatest increase in the rate of adsorption of metal ions on sawdust was observed for pH changes from 2 to 5. An empirical relationship has been obtained to predict the percentage Nickel (II) removal at any time for known values of sorbent and initial sorbate concentration. The experimental results provided evidence for chelation ion exchange as the major adsorption mechanisms for binding metal ions to the sawdust. The adsorbent can be effectively regenerated using 0.1 M strong acid and reused.

The relationship between the cellular Ca2+ signal and exocytic vesicle fusion is a key determinant of the regulation of the kinetics and magnitude of the secretory response. Here, we have investigated secretion in epithelial cells where the exocytic response is controlled by Ca2+ release from intracellular Ca2+ stores (Ueda & Petersen 1977). Using live-cell 2-photon microscopy, identifying each individual exocytic fusion event (Thorn et al., 2004) and recording cytosolic Ca2+ signals with Fura-2 (Nemoto et al., 2001), we show no evidence for spatial clustering of exocytosis; indeed exocytosis is actually specifically excluded from sites of Ca2+ release hot-spots. Consistent with this data, loading the cells with EGTA potently blocks exocytosis. These results indicate that the control of exocytosis, triggered by Ca2+ release from stores, is through the regulation of cytosolic Ca2+ concentrations within large volume microdomains.Nemoto, T., Kimura, R., Ito, K., Tachikawa, A., Miyashita, Y., Iino, M. & Kasai, H. (2001) Nature Cell Biol.3: 253-258.Thorn, P., Fogarty, K.E. & Parker, I. (2004) Proceedings National Academy Science 101:6774-6779Ueda N. & Petersen OH (1997) Pflugers Archiv370: 179-183

Measurement of product ratios from dediazoniation reactions in acidic aqueous solutions of cationic micelles provides a new method for determining the relative interfacial quantities of weak nucleophiles at the surface of association colloids. We determined product yields by HPLC from decomposition of hydrophobic derivatives of benzenediazonium salts in aqueous solutions of cetyltrimethyl-ammonium halide, CTAX (X =C1− and Br−) micelles and 0.1 M HX with added NaX at 40°C. Estimates of the ion exchange constant, Kcl Br≃3, obtained from product ratios in the presence of mixtures of the two ions are independent of added salt, but decrease to about 2 at high Br- mole ratio. Preliminary results in CTABr micelles show that the method can be used with microemulsions and that added BuOH reduces the quantity of interfacial Br− as expected, but surprisingly not interfacial water. Generous financial support was provided by NIH, NSF-U.S. Brazil Cooperative Program, PRF, Research Corporation and The Research Council and BRSG Fund of Rutgers University.

67. Bits and pieces, 27. It is important to realize the subtly of the computation and to note that computers have some limitations.