Allyl piperidine-1-carbodiothioate and benzyl 1H-imidazole 1 carbodithioate: two potential agents to combat against mycobacteria.

AIMS: The emergence of multidrug resistant strains of Mycobacterium tuberculosis has made tuberculosis more difficult to manage clinically. With the aim of obtaining new and effective anti-mycobacterial agent(s), this study investigated the anti-mycobacterial activity of several imidazole and piperidine derivatives. METHODS AND RESULTS: Towards obtaining new anti-mycobacterial agents, Mycobacterium smegmatis cells were treated with different compounds for their growth inhibitory activity. Among these, benzyl 1H-imidazole-1-carbodithioate and allyl piperidine-1-carbodiothioate exhibited better inhibition than the others. Thereafter, anti-biofilm property of these two was examined by treating M. smegmatis with these agents before and after the formation of biofilm. The result showed that both the compounds at their sublethal dose inhibited the formation of biofilm as well as dispersed preformed biofilm. Consistently, they augmented the activity of isoniazid or rifampicin against biofilm-encapsulated cells. MTT assay was performed to examine the toxic effects of this combinatorial therapy on different cell lines. Results exhibited a low cytotoxicity for this combinatorial treatment. The activity of these two was also verified against dormant mycobacterial cells and was found to be effective. CONCLUSION: The present study identified two compounds that exhibited anti-mycobacterial activities against both planktonic and dormant cells. These two also exhibited anti-biofilm activity at their sublethal dose and augmented the activity of isoniazid and rifampicin against biofilm encapsulated cells. SIGNIFICANCE AND IMPACT OF THE STUDY: The current study provides two new agents that have the potential to be used in anti-mycobacterial therapy and may help in public health management.

First Observation of Multiple Transverse Wobbling Bands of Different Kinds in

We report the first observation of two wobbling bands in ^{183}Au, both of which were interpreted as the transverse wobbling (TW) band but with different behavior of their wobbling energies as a function of spin. It increases (decreases) with spin for the positive (negative) parity configuration. The crucial evidence for the wobbling nature of the bands, dominance of the E2 component in the ΔI=1 transitions between the partner bands, is provided by the simultaneous measurements of directional correlation from the oriented states ratio and the linear polarization of the γ rays. Particle rotor model calculations with triaxial deformation reproduce the experimental data well. A value of spin, I_{m}, has been determined for the observed TW bands below which the wobbling energy increases and above which it decreases with spin. The nucleus ^{183}Au is, so far, the only nucleus in which both the increasing and the decreasing parts are observed and thus gives the experimental evidence of the complete transverse wobbling phenomenon.

Generation of ß cell-specific human cytotoxic T cells by lentiviral transduction and their survival in immunodeficient human leucocyte antigen-transgenic mice.

Several ß cell antigens recognized by T cells in the non-obese diabetic (NOD) mouse model of type 1 diabetes (T1D) are also T cell targets in the human disease. While numerous antigen-specific therapies prevent diabetes in NOD mice, successful translation of rodent findings to patients has been difficult. A human leucocyte antigen (HLA)-transgenic mouse model incorporating human ß cell-specific T cells might provide a better platform for evaluating antigen-specific therapies. The ability to study such T cells is limited by their low frequency in peripheral blood and the difficulty in obtaining islet-infiltrating T cells from patients. We have worked to overcome this limitation by using lentiviral transduction to 'reprogram' primary human CD8 T cells to express three T cell receptors (TCRs) specific for a peptide derived from the ß cell antigen islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP265-273 ) and recognized in the context of the human class I major histocompatibility complex (MHC) molecule HLA-A2. The TCRs bound peptide/MHC multimers with a range of avidities, but all bound with at least 10-fold lower avidity than the anti-viral TCR used for comparison. One exhibited antigenic recognition promiscuity. The ß cell-specific human CD8 T cells generated by lentiviral transduction with one of the TCRs released interferon (IFN)-γ in response to antigen and exhibited cytotoxic activity against peptide-pulsed target cells. The cells engrafted in HLA-A2-transgenic NOD-scid IL2rγ(null) mice and could be detected in the blood, spleen and pancreas up to 5 weeks post-transfer, suggesting the utility of this approach for the evaluation of T cell-modulatory therapies for T1D and other T cell-mediated autoimmune diseases.

Measurement of energy and directional distribution of neutron ambient dose equivalent for the 7Li(p,n)7Be reaction.

Double differential neutron fluence distributions were measured in the 7Li(p,n)7Be reaction for proton beam energies 7, 9 and 12 MeV. Seven liquid scintillator based detectors were employed to measure neutron fluence distributions using the Time of Flight technique. Neutron ambient dose equivalents were determined from the measured fluence distribution using ICRP (International Commission on Radiological Protection) recommended fluence to dose equivalent conversion coefficients. Neutron dose equivalents were also measured using a conventional BF3 detector based REM counter. Ambient dose equivalent measured by the REM counter is found to be in agreement with that determined from the neutron fluence spectra within their uncertainties. Angular distributions of the ambient dose equivalents were also determined from the measured fluence distributions at different angles.

Relationship between lymph node metastases and histopathological parameters in carcinoma cervix: a multivariate analysis.

A retrospective review of 360 cases of carcinoma of the cervix with clinical stage IB and IIA who had undergone radical hysterectomy and pelvic node dissection between 2000 and 2005 was carried out. Lymph node metastasis was present in 79/360 patients (21.9%). LVSI positivity, full thickness stromal invasion, involvement of the uterine isthmus, positive parametrium, positive vaginal margins, involvement of uterine corpus was seen in: 25.3% and 9.2% (p < 0.001); 63% and 32% (p < 0.001); 32.9% and 13.8% (p < 0.001);15.2% and 5% (p < 0.004); 24% and 14.2% (p < 0.005); 17.7% and 13.8% (p = 0.11) of the patients, with and without lymph node metastasis, respectively. The tumour size was <4 cm in 50.6% and 58.3% and >4 cm in 49.3% and 41.6% of the patients, with and without lymph node metastasis, respectively (p = 0.22), which was statistically not significant. In the majority of patients, the histopathology type was squamous cell carcinoma in both the groups. In patients with lymph node metastases 79.7% had grade III tumour as compared with 69.5% in patients without lymph node metastases (p = 0.19). Multiple logistic regression indicated that only lymphovascular space involvement and full thickness stromal invasion were statistically significant (p < 0.001 and p < 0.002, respectively) for lymph node metastasis.

Notch1 regulates the functional contribution of RhoC to cervical carcinoma progression.

BACKGROUND: The role of Notch signalling in human epithelial cancers is of immense interest. In this study, we examine the interplay between Notch signalling and RhoC, a well-established molecular factor in metastasis. By linking the function of Notch and RhoC, we further strengthen the notion that there is a pro-oncogenic role of Notch signalling in human cervical cancers. METHODS: RhoC protein expression in cervical carcinoma cell lines was assessed by western blotting. Using CaSki and SiHa cells (cervical carcinoma cells lines), we show that RhoC contributes to wound healing, invasion and migration, anoikis resistance, colony formation, in vitro tube formation and tumour formation. Immunohistochemical studies were carried out to assess the co-expression of RhoC, pAkt and Notch1 in clinical sections. RESULTS: An assessment of the changes associated with epithelial-to-mesenchymal transition (EMT) shows that both Notch1 and RhoC have similar phenotypic contribution to EMT. Rho activity assessment on Notch1 inhibition with DAPT shows decreased RhoC activity. We further show that constitutively active RhoC rescues the phenotypic effect of Notch1 inactivation, and a comparison of Notch1 with RhoC expression shows an overlap between the two proteins in the same areas of the tissue. CONCLUSION: This study has provided evidence to suggest that RhoC is an effector of Notch1 in cervical carcinoma.

The immunotherapeutic potential of dendritic cells in type 1 diabetes.

Type 1 diabetes is an autoimmune disease characterized by destruction of the pancreatic islet beta cells that is mediated primarily by T cells specific for beta cell antigens. Insulin administration prolongs the life of affected individuals, but often fails to prevent the serious complications that decrease quality of life and result in significant morbidity and mortality. Thus, new strategies for the prevention and treatment of this disease are warranted. Given the important role of dendritic cells (DCs) in the establishment of peripheral T cell tolerance, DC-based strategies are a rational and exciting avenue of exploration. DCs employ a diverse arsenal to maintain tolerance, including the induction of T cell deletion or anergy and the generation and expansion of regulatory T cell populations. Here we review DC-based immunotherapeutic approaches to type 1 diabetes, most of which have been employed in non-obese diabetic (NOD) mice or other murine models of the disease. These strategies include administration of in vitro-generated DCs, deliberate exposure of DCs to antigens before transfer and the targeting of antigens to DCs in vivo. Although remarkable results have often been obtained in these model systems, the challenge now is to translate DC-based immunotherapeutic strategies to humans, while at the same time minimizing the potential for global immunosuppression or exacerbation of autoimmune responses. In this review, we have devoted considerable attention to antigen-specific DC-based approaches, as results from murine models suggest that they have the potential to result in regulatory T cell populations capable of both preventing and reversing type 1 diabetes.

X-ray diffraction measurements of Mo melting to 119 GPa and the high pressure phase diagram.

In this paper, we report angle-dispersive X-ray diffraction data of molybdenum melting, measured in a double-sided laser-heated diamond-anvil cell up to a pressure of 119 GPa and temperatures up to 3400 K. The new melting temperatures are in excellent agreement with earlier measurements up to 90 GPa that relied on optical observations of melting and in strong contrast to most theoretical estimates. The X-ray measurements show that the solid melts from the bcc structure throughout the reported pressure range and provide no evidence for a high temperature transition from bcc to a close-packed structure, or to any other crystalline structure. This observation contradicts earlier interpretations of shock data arguing for such a transition. Instead, the values for the Poisson ratios of shock compressed Mo, obtained from the sound speed measurements, and the present X-ray evidence of loss of long-range order suggest that the 210 GPa (approximately 4100 K) transition in the shock experiment is from the bcc structure to a new, highly viscous, structured melt.

ArrayExpress--a public repository for microarray gene expression data at the EBI.

ArrayExpress is a public repository for microarray data that supports the MIAME (Minimum Information About a Microarray Experiment) requirements and stores well-annotated raw and normalized data. As of November 2004, ArrayExpress contains data from approximately 12,000 hybridizations covering 35 species. Data can be submitted online or directly from local databases or LIMS in a standard format, and password-protected access to prepublication data is provided for reviewers and authors. The data can be retrieved by accession number or queried by various parameters such as species, author and array platform. A facility to query experiments by gene and sample properties is provided for a growing subset of curated data that is loaded in to the ArrayExpress data warehouse. Data can be visualized and analysed using Expression Profiler, the integrated data analysis tool. ArrayExpress is available at http://www.ebi.ac.uk/arrayexpress.

Ferulic acid esterase production by Streptomyces sp.

Studies were carried out on ferulic acid esterase production using a culture of Streptomyces S10. In optimized condition, enzyme yield was 2.0 mU/ml in MBS medium, containing 1.5% de-starched wheat bran at 30 degrees C and initial pH 6.5 under agitated submerged culture.

Septic abortions: a descriptive study in a teaching hospital at North Bengal, Darjeeling.

A hospital based cross sectional study for one year done among 57 diagnosed and admitted septic abortion cases, revealed that 71.9% septic abortions were performed by untrained persons, 63.2% mothers were illiterate, 22.8% mothers were in adolescent age group. Exploratory Laparotomy was needed in 46.3% cases and important complications for referring were severe anaemia and septic shock.

Weighted scale-free networks in Euclidean space using local selection rule.

A spatial scale-free network is introduced and studied, whose motivation originated in the growing Internet as well as airport networks. We argue that in these real-world networks a new node necessarily selects one of its neighboring local nodes for connection and is not controlled by preferential attachment as in the Barabási-Albert (BA) model. This observation is mimicked in our model where the nodes pop up at randomly located positions in the Euclidean space and are connected to one end of the nearest link. In spite of this crucial difference it is observed that the leading behavior of our network is like that of the BA model. Defining the link weight as an algebraic power of its Euclidean length, the weight distribution and the nonlinear dependence of the nodal strength on the degree are analytically calculated. It is claimed that a power law decay of the link weights with time ensures such nonlinear behavior. Switching off the Euclidean space from the same model yields a much simpler definition of the BA model where numerical effort grows linearly with N.

A consortium of non-rhizobial endophytic microbes from Typha angustifolia functions as probiotic in rice and improves nitrogen metabolism.

Endophytic microbes isolated from plants growing in nutrient-deficient environments often possess properties that improve nutrition of agriculturally important plants. A consortium of non-rhizobial endophytic microbes isolated from a macrophyte Typha angustifolia growing in the marginal wetlands associated with a Uranium mine was characterized for their beneficial effect on rice and the mechanisms of growth promotion were investigated. The microbes were identified and characterized for their potential plant growth promoting (PGP) properties. Effect of these microbes on nitrogen (N)-metabolism of rice was tested as Typha endophytes were predominantly (N)-fixing. Relative N-use efficiency and expression of genes involved in N-uptake and assimilation were investigated in treated plants. Evidence of horizontal gene transfer (HGT) of dinitrogen reductase gene was observed within the consortium from a Pseudomonas stutzeri strain. The consortium behaved as plant probiotic and showed substantial growth benefits to Typha, their natural host as well as to rice. Typha endophytes colonized rice endosphere significantly increasing biomass, shoot length and chlorophyll content in rice plants both under N-sufficient and N-deficient conditions. N-uptake and assimilation genes were upregulated in plants treated with the endophytes even after three weeks post infection. Our results suggested, HGT of nitrogen-fixation trait to be highly prevalent among endophytes isolated from nutrient-poor habitats of the uranium mine. A long-term nitrogen deficiency response in the treated plants was elicited by the consortium improving N-uptake, assimilation and relative N-use efficiency of rice plants. This appeared to be at least one of the main strategies of plant growth promotion.

Constitutive activation of FLT3 stimulates multiple intracellular signal transducers and results in transformation.

Aberrant expression of FLT3 has been found in most cases of B-lineage ALL and AML, and subsets of T cell ALL, CML in blast crisis and CLL. In 20% of patients with AML the receptor has small internal tandem duplications of the juxtamembrane region which appear to contitutively activate the receptor. To investigate whether FLT3 activation could play a role in leukemia, we generated a constitutively activated FLT3 by fusing its cytoplasmic domain to the helix-loop-helix domain of TEL in analogy to the fusion that occurs with TEL-PDGFR in CMML. In vitro translation assays demonstrated oligomerization and intrinsic tyrosine kinase activity of the TEL-FLT3 chimeric receptor. Constitutively activated TEL-FLT3 conferred IL-3 independence and long-term proliferation to transfected Ba/F3 cells. Immunoblot analyses showed that JAK 2, STAT 3, STAT 5a, STAT 5b and CBL were tyrosine-phosphorylated in TEL-FLT3 expressing Ba/F3 cells in the absence of IL-3. These data suggest a possible role for the JAK/STAT pathway in FLT3 signaling. Transplantation of TEL-FLT3 expressing Ba/F3 cells into syngeneic mice caused mortality in all mice by 3 weeks after injection. Histopathologic analysis demonstrated a massive infiltration of mononuclear cells in the liver, spleen and bone marrow. The mimicking of naturally occurring TEL fusions provides an approach to assess aspects of the biology of activated FLT3, or other receptor-type tyrosine kinases (RTKs) in leukemic transformation.

Remodeling of the plasma membrane after stimulation of neutrophils with f-Met-Leu-Phe and dihydrocytochalasin B: identification of membrane subdomains containing NADPH oxidase activity.

Superoxide (O2-) production by neutrophils stimulated with chemotactic peptides [e.g., formylmethionyl-leucyl-phenylalanine (fMLP)] is transient but increases in rate and duration after pretreatment of the cells with dihydrocytochalasin B (dhCB), suggesting a possible role for the plasma membrane and membrane skeleton in the regulation of the O2- generating system. Analysis of plasma membrane isolated from these cells by isopycnic sucrose density gradient sedimentation showed that there were no significant variation in the distribution of plasma membrane markers between control and dhCB-treated cells, whereas a significant redistribution of plasma membrane markers was observed in dhCB + fMLP-treated cells. Instead of sedimenting to 31-35% sucrose, as in the former two groups, plasma membrane markers were broadly distributed over 25-50% sucrose in the dhCB + fMLP-treated cells. In addition, approximately 80% degranulation was achieved in these cells, whereas little granule release (< 5%) was observed in control and dhCB-treated cells. Analysis of the gradient fractions for membrane skeletal (actin and fodrin) and NADPH oxidase (cytochrome b and p47-phox) components in dhCB + fMLP-treated cells demonstrated that the distribution of fodrin, actin, cytochrome b, and p47-phox followed the broad distribution of plasma membrane markers, with an overall eightfold increase in membrane-associated actin. Despite the broad redistribution of plasma membrane markers, the distribution of O2- generating activity remained confined to a narrower peak at approximately 50% sucrose. These results demonstrate that a heterogeneous surface membrane of higher density with a differential distribution of proteins and O2- generating activity are created after dhCB + fMLP treatment; however, domain structure is conserved in the new membrane, with only a subfraction of the reorganized plasma membrane containing all of the components necessary for active O2- generation. Our results support a role for plasma membrane lateral organization and participation of the membrane skeleton in the regulation of the O2- generating system.

Phase transition in a directed traffic flow network.

The generic feature of traffic in a network of flowing electronic data packets is a phase transition from a stationary free-flow phase to a continuously growing congested nonstationary phase. In the most simple network of directed oriented square lattice we have been able to observe all crucial features of such flow systems having nontrivial critical behavior near the critical point of transition. The network here is in the shape of a square lattice and data packets are randomly posted with a rate rho at one side of the lattice. Each packet executes a directed diffusive motion toward the opposite boundary where it is delivered. Packets accumulated at a particular node form a queue and a maximum of m such packets randomly jump out of this node at every time step to its neighbors on a first-in-first-out basis. The phase transition occurs at rho(c) = m. The distribution of travel times through the system is found to have a log-normal behavior and the power spectrum of the load time series shows 1/f-like noise similar to the scenario of Internet traffic.

Comparisons of alloreactive potential of clinical hematopoietic grafts.

BACKGROUND: We sought to compare the immunoreactive potential of human cord blood (CB) versus normal adult bone marrow (BM) versus mobilized blood (peripheral blood stem cells; PBSC) from cancer patients. METHODS: Forty mice were randomized to receive a range of doses of T cell-replete cell preparations from one of the above three cell sources. Twenty-eight control mice underwent transplantation with T cell-depleted cells. Mice were observed for 60 days for the development of fatal xenogeneic graft-versus-host disease-like syndrome (GVHDLS). RESULTS: For the mice that had received T cell-replete grafts of CB or BM or PBSC, the duration of GVHDLS-free survival of the chimeras was inversely proportional to the number of T cells transplanted. After adjustment for the number of T cells transplanted, the relative hazard of developing fatal GVHDLS was 62-fold higher for PBSC and 210-fold higher for BM as compared with CB. Flow cytometric and histologic analyses of selected chimeras that died of GVHDLS showed extensive proliferation of human T cells in multiple organs. In contrast, mice that survived to day 60 were engrafted with human myeloid and B lymphoid cells. CONCLUSIONS: The immunoreactive potential, as measured by this in vivo assay, differed among clinical grafts: BM > PBSC > CB.

Squamous cell carcinoma arising in endometriosis of the ovary.

Squamous cell carcinoma arising in ovarian endometriosis is extremely rare, but a case is reported in a 62 year old woman. If such an ovarian tumour is discovered evidence of teratoma, Brenner tumour, or endometriosis should be sought. Extensive sampling is mandatory as the developing tumour often overgrows the foci of endometriosis, thus obliterating evidence of its origin.