Advanced hybrid attention-based deep learning network with heuristic algorithm for adaptive CT and PET image fusion in lung cancer detection.

Lung cancer is one of the most deadly diseases in the world. Lung cancer detection can save the patient's life. Despite being the best imaging tool in the medical sector, clinicians find it challenging to interpret and detect cancer from Computed Tomography (CT) scan data. One of the most effective ways for the diagnosis of certain malignancies like lung tumours is Positron Emission Tomography (PET) imaging. So many diagnosis models have been implemented nowadays to diagnose various diseases. Early lung cancer identification is very important for predicting the severity level of lung cancer in cancer patients. To explore the effective model, an image fusion-based detection model is proposed for lung cancer detection using an improved heuristic algorithm of the deep learning model. Firstly, the PET and CT images are gathered from the internet. Further, these two collected images are fused for further process by using the Adaptive Dilated Convolution Neural Network (AD-CNN), in which the hyperparameters are tuned by the Modified Initial Velocity-based Capuchin Search Algorithm (MIV-CapSA). Subsequently, the abnormal regions are segmented by influencing the TransUnet3+. Finally, the segmented images are fed into the Hybrid Attention-based Deep Networks (HADN) model, encompassed with Mobilenet and Shufflenet. Therefore, the effectiveness of the novel detection model is analyzed using various metrics compared with traditional approaches. At last, the outcome evinces that it aids in early basic detection to treat the patients effectively.

Ultrasensitive and reversible NO2 gas sensor based on SnS2/TiO2 heterostructures for room temperature applications.

Nitrogen dioxide (NO2) is one of the toxic gases produced by chemical industries, power plants, and vehicles. In this work, we demonstrate an inexpensive sensing platform for NO2 detection at room temperature (RT-32 °C) based on a charge transfer mechanism. Three-dimensional hierarchical SnS2 and SnS2/mesoporous TiO2 nanocomposites were synthesized via the solvothermal method. SnS2/20 wt% mesoporous TiO2 nanocomposites sample showed 245.4% enhanced response compared to pristine SnS2. The fabricated device exhibits excellent selectivity among all other interfering gases with one-month stability. The rapid response and enhanced response achieved were obtained for the minimum concentration of 2 ppm NO2. The formation of heterojunction between SnS2 and mesoporous TiO2 has a synergetic effect, providing more active sites and porous structures for the detection of NO2 gas molecules.

Vascular Endothelial Growth Factor Receptor, fms-Like Tyrosine Kinase-1 (Flt-1), as a Novel Binding Partner for SARS-CoV-2 Spike Receptor-Binding Domain.

Angiotensin-converting enzyme 2 (ACE2) and neuropilin 1, a vascular endothelial growth factor (VEGF) receptor, were identified to bind to the SARS-CoV-2 spike receptor-binding domain (spike RBD). In silico analysis based on 3D structure, multiple sequence alignment, and molecular docking of second domain of soluble Flt-1 (sFlt-1) and spike RBD revealed structural similarities, sequence homology, and protein-protein interaction. Interaction and binding of recombinant spike RBD (rspike RBD) and recombinant sFlt-1 (rsFlt-1) in vitro induced a conformational change, as revealed by spectrofluorimetric data, with increased fluorescence intensity in emission spectra as compared to either of the proteins alone. Results on ELISA confirmed the binding and cross-reactivity of rspike-RBD and rsFlt-1 as determined by using either specific antibodies towards each protein or immunized human serum. We found that polyclonal or monoclonal anti-spike RBD antibodies can recognize either rsFlt-1 or rspike RBD, showing cross-reactivity for the two proteins in a dose-dependent binding response. Recognition of bound rspike RBD or rsFlt-1 by anti-Flt-1 or anti-spike RBD antibodies, respectively, as observed by immunoblotting, further confirmed interaction between the two proteins. Immunoprecipitation and immunoblot analysis demonstrated the identification of rspike RBD binding to the Flt-1 receptor on A549 cells. Further, the binding of rspike RBD to Flt-1 receptor was shown using immunofluorescence on 2D-culture or 3D-spheroid of MDA-MB-231 cells, which over-express Flt-1 receptor. Together, our study concludes that the Flt-1 receptor is a novel binding partner for SARS-CoV-2 spike RBD.

Overview of diagnostics on a small-scale RF source for fusion (ROBIN) and the one planned for the diagnostic beam for ITER.

India is responsible for the supply of diagnostic neutral beam systems for ITER to diagnose its helium ash during the deuterium-tritium plasma phase using the charge exchange recombination spectroscopy technique. Considering the many first of its kind in terms of technologies and beam development aspects, ITER Indian domestic agency has adopted a strategy of developing the technology and beam experimentation in parallel. On the beam development front three test beds, namely, the ROBIN (Rf Operated Beam source in India for Negative ion research), the TWIN (TWo rf driver-based Indigenously built Negative ion source), and the INTF (INdian Test Facility) are presently in their various phases of operation, optimization, and setting up at IPR, respectively. Experiments related to plasma production, beam production, and acceleration up to 30 keV in volume and surface mode have been performed on ROBIN. The maximum negative hydrogen ion current density to a tune of 27 mA/cm2 is obtained in the surface mode with Cs injection. Optimal source performance requires optimal surface conditions, minimum impurities, careful characterization of the plasma, cesium feed and its redistribution, and optimal wall temperatures of the surfaces of the plasma box and the plasma grid. A combination of probe, optical, vacuum, laser based, electrical, and calorimetric diagnostic measurements enables such a control. At ROBIN, the above diagnostics are being used regularly. The operational and diagnostic experiences on ROBIN shall provide the desired experience and database for operations of TWIN and INTF in the coming years. A large number of conventional and advanced diagnostic techniques are used for plasma and beam characterization. These diagnostics are suitable not only to detect and understand the plasma but also for studies related to impurity evolution. The temporal evolution of impurities significantly impacts the plasma and beam properties. The studies help in establishing correlations between physical parameters and operational parameters to optimize the source performance ensuring adequate safety and investment protection. This paper will present a brief overview of various diagnostics implemented, lessons learned, and the results obtained from ROBIN. In addition, an outline of the diagnostics planned for INTF based on the experience and understandings developed during the present experiments on ROBIN and TWIN and considering the requirements of large systems shall be discussed.

Characterization and Cytotoxicity of Pseudomonas Mediated Rhamnolipids Against Breast Cancer MDA-MB-231 Cell Line.

A biosurfactant producing bacterium was identified as Pseudomonas aeruginosa DNM50 based on molecular characterization (NCBI accession no. MK351591). Structural characterization using MALDI-TOF revealed the presence of 12 different congeners of rhamnolipid such as Rha-C8-C8:1, Rha-C10-C8:1, Rha-C10-C10, Rha-C10-C12:1, Rha-C16:1, Rha-C16, Rha-C17:1, Rha-Rha-C10:1-C10:1, Rha-Rha-C10-C12, Rha-Rha-C10-C8, Rha-Rha-C10-C8:1, and Rha-Rha-C8-C8. The radical scavenging activity of rhamnolipid (DNM50RL) was determined by 2, 3-diphenyl-1-picrylhydrazyl (DPPH) assay which showed an IC50 value of 101.8 µg/ ml. The cytotoxic activity was investigated against MDA-MB-231 breast cancer cell line by MTT (4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide) assay which showed a very low IC50 of 0.05 µg/ ml at 72 h of treatment. Further, its activity was confirmed by resazurin and trypan blue assay with IC50 values of 0.01 µg/ml and 0.64 µg/ ml at 72 h of treatment, respectively. Thus, the DNM50RL would play a vital role in the treatment of breast cancer targeting inhibition of p38MAPK.

Interface enriched highly interlaced layered MoS2/NiS2 nanocomposites for the photocatalytic degradation of rhodamine B dye.

In the past few decades, air and water pollution by organic dyes has become a serious concern due to their high toxicity. Removal of these organic dyes from polluted water bodies is a serious environmental concern and the development of new advanced photocatalytic materials for decomposing organic dyes can be a good solution. In this work, layered molybdenum disulfide/nickel disulfide (MoS2/NiS2) nanocomposites with various NiS2 content was synthesized by a one-step hydrothermal method using citric acid as a reducing agent. The X-ray diffraction pattern shows the hexagonal and cubical crystal structure of MoS2 and NiS2, respectively. Morphological analysis confirms the formation of MoS2/NiS2 nanosheets. The elemental composition of the samples was carried out by XPS, which shows a significant interaction between NiS2 and MoS2. The photocatalytic performance of MoS2/NiS2 nanocomposites was studied by the degradation of rhodamine B (RhB). Ni-4 sample shows higher photocatalytic activity with a maximum degradation of 90.61% under visible light irradiation for 32 min.

Comprehensive review of guidelines to practice prosthodontic and implant procedures during COVID-19 pandemic.

OBJECTIVES: To present a comprehensive review of current literature available on Corona virus disease and dentistry, modifications required in dental and laboratory settings; and recommended disinfection protocols in current scenario. Special emphasis has been given to discuss guidelines for handling different prosthodontic procedures and implications of this pandemic on prosthodontic practice, education and research. MATERIALS AND METHODS: Relevant literature pertaining to COVID-19 and dentistry was scrutinized on electronic search engines including PubMed, EMBASE and Cochrane. Guidelines given by various organizations, institutions, national and international regulatory bodies and Indian Prosthodontic Society were also consulted to gather information pertaining to the objectives of our review. RESULTS: and observations: A total of 160 articles including cross-sectional studies, in vitro study, narrative reviews, letters to the editor and opinions were found to be relevant in accordance with our search strategy. Documented literature revealed that Covid-19 pandemic has culminated in serious clinical, financial and psychological implications in the field of dentistry. Certain steps such as adoption of teledentistry, judicious use of protective equipment, use of rubber dams, pre-procedural rinses have been suggested unanimously. However, there is a dearth of evidence-based recommendations in literature. Data regarding consequences of delaying prosthodontic procedures and patients' perspectives is also sparse. CONCLUSION: The COVID-19 necessitates the need to adopt a balanced approach while treating patients and safeguarding the dental professionals at the same time. Risk-benefit ratio has to be assessed along with stringent following of guidelines and disinfection protocols to combat this unprecedented situation.

Synthesis and Screening of Pro-apoptotic and Angio-inhibitory Activity of Novel Benzisoxazole Derivatives both In Vitro and In Vivo.

BACKGROUND: Triple Negative Breast Cancer (TNBC) tends to be more aggressive than other types of breast cancer. Resistance to chemotherapy is a major obstacle hence there is a significant need for new antineoplastic drugs with multi-target potency. Numerous Benzoisoxazole moieties have been found to possess a broad spectrum of pharmacological activities. In the present study, we have synthesized 9 novel derivatives of Benzisoxazole 7(a-i) and screened them for their biological potential. METHODS: Chemical synthesis, Mass spectrometry (HRMS), cell proliferation and cytotoxicity assay, wound healing assay, flow cytometry and nuclear staining. Angio-inhibitory activity assessed by corneal micropocket assay and in vivo peritoneal angiogenesis assay. RESULTS: The Benzisoxazole derivatives 7(a-i) were synthesized and screened for their biological potency by both in vitro and in vivo experimental models. Among the series, compound 3-(1-((3-(3(Benzyloxy)-4-methoxyphenyl)- 4,5-dihydroisoxazole-5-yl)methyl)piperidine-4-yl)6-fluorobenzo[d] isoxazole (7e) was found to be most promising, with an average IC50 value of 50.36 ± 1.7 µM in MTT assay and showed 81.3% cell death. The compound 7e also showed 60-70% inhibition on a recombinant Metastasis-Associated protein (MTA1) induced proliferation and cell migration in MDAMB-231 cells, which is known to play a major role in angiogenesis. The anti-tumour studies inferred the regression of tumour activity. This was due to inhibition of neovascularization and evoking apoptosis process as assessed by corneal vascularization, peritoneal angiogenesis and apoptotic hallmarks in 7e treated cells. CONCLUSION: These findings not only show the biological efficacy of compound 7e but it is also an effective beginning to explore the mechanism of metastasis and cancer therapy strategy targeting MTA1. The observed biological activity makes compound 7e an attractive drug candidate.

A new pyrrole based small molecule from Tinospora cordifolia induces apoptosis in MDA-MB-231 breast cancer cells via ROS mediated mitochondrial damage and restoration of p53 activity.

Approximately 15% of globally diagnosed breast cancers are designated as triple negative breast cancer (TNBC). In this study, we investigated the effect of the natural compound, Bis(2- ethyl hexyl) 1H-pyrrole-3,4-dicarboxylate (TCCP), purified from Tinospora cordifolia on MDA-MB-231, a TNBC cell line. The pro-apoptotic nature of TCCP on MDA-MB-231 was determined by assessing various apoptotic markers. ROS generation, intracellular calcium, mitochondrial membrane potential (ΔΨm), MPTP, cardiolipin peroxidation and caspase activity were determined fluorometrically. BAX, BCL-2, cytochrome c, caspases, and p53 protein expressions were determined by immunoblotting. Further, the effect of TCCP on DNA and cell death was determined by DNA fragmentation assay, annexin-V staining, and cell cycle analysis. TCCP treatment caused endogenous ROS generation, increase in intracellular calcium and phosphorylation of p53 in a concentration-dependent manner, which was reverted upon pre-treatment with pifithrin-µ. This led to the downstream altered expression of Bcl-2 and Bax proteins, mitochondrial membrane depolarization, MPTP, and cardiolipin peroxidation. TCCP induced cytochrome c release into the cytosol, caspase activation, ultimately resulting in DNA fragmentation. Further, induction of apoptosis and morphological alterations were evident from the phosphatidylserine externalization and increase in sub G1 population. The in vivo Ehrlich ascites tumor (EAT) mouse study revealed the effectiveness of TCCP in reducing the tumor burden and resulted in a ~2 fold increase in mice survival with minimal hepato-renal toxicity. Overall, TCCP was shown to be efficient in inducing ROS and mitochondrial-mediated apoptosis by restoring p53 activity in MDA-MB-231 cells and also induced EAT cell death in vivo thereby inhibiting tumor proliferation.

A pyrrole-based natural small molecule mitigates HSP90 expression in MDA-MB-231 cells and inhibits tumor angiogenesis in mice by inactivating HSF-1.

Heat shock proteins (HSPs), molecular chaperones, are crucial for the cancer cells to facilitate proper functioning of various oncoproteins involved in cell survival, proliferation, migration, and tumor angiogenesis. Tumor cells are said to be "addicted" to HSPs. HSPs are overexpressed in many cancers due to upregulation of transcription factor Heat-shock factor 1 (HSF-1), the multifaceted master regulator of heat shock response. Therefore, pharmacological targeting of HSPs via HSF-1 is an effective strategy to treat malignant cancers like triple negative breast cancer. In the current study, we evaluated the efficacy of a pyrrole derivative [bis(2-ethylhexyl)1H-pyrrole-3,4-dicarboxylate], TCCP, purified from leaves of Tinospora cordifolia for its ability to suppress heat shock response and angiogenesis using MDA-MB-231 cells and the murine mammary carcinoma: Ehrlich ascites tumor model. HSP90 was downregulated by TCCP by inactivation of HSF-1 resulting in inhibition of tumor cell proliferation, VEGF-induced cell migration, and concomitant decrease in tumor burden and neo-angiogenesis in vivo. The mechanism of suppression of HSPs involves inactivation of PI3K/Akt and phosphorylation on serine 307 of HSF-1 by the activation of ERK1. HSF-1 and HSP90 and 70 localization and expression were ascertained by immunolocalization, immunoblotting, and qPCR experiments. The anti-angiogenic effect of TCCP was studied in vivo in tumor-bearing mice and ex vivo using rat corneal micro-pocket assay. All the results thus corroborate the logic behind inactivating HSF-1 using TCCP as an alternative approach for cancer therapy.

Suppression of VEGF-induced angiogenesis and tumor growth by Eugenia jambolana, Musa paradisiaca, and Coccinia indica extracts.

CONTEXT: Abnormal angiogenesis and evasion of apoptosis are hallmarks of cancer. Accordingly, anti-angiogenic and pro-apoptotic therapies are effective strategies for cancer treatment. Medicinal plants, namely, Eugenia jambolana Lam. (Myrtaceae), Musa paradisiaca L. (Musaceae), and Coccinia indica Wight & Arn. (Cucurbitaceae), have not been greatly investigated for their anticancer potential. OBJECTIVE: We investigated the anti-angiogenic and pro-apoptotic efficacy of ethyl acetate (EA) and n-butanol (NB) extracts of E. jambolana (seeds), EA extracts of M. paradisiaca (roots) and C. indica (leaves) with respect to mammary neoplasia. MATERIALS AND METHODS: Effect of extracts (2-200 µg/mL) on cytotoxicity and MCF-7, MDA-MB-231 and endothelial cell (EC) proliferation and in vitro angiogenesis were evaluated by MTT, 3[H]thymidine uptake and EC tube formation assays, respectively. In vivo tumour proliferation, VEGF secretion and angiogenesis were assessed using the Ehrlich ascites tumour (EAT) model followed by rat corneal micro-pocket and chicken chorioallantoic membrane (CAM) assays. Apoptosis induction was assessed by morphological and cell cycle analysis. RESULTS: EA extracts of E. jambolana and M. paradisiaca exhibited the highest cytotoxicity (IC50 25 and 60 µg/mL), inhibited cell proliferation (up to 81%), and tube formation (83% and 76%). In vivo treatment reduced body weight (50%); cell number (16.5- and 14.7-fold), secreted VEGF (∼90%), neoangiogenesis in rat cornea (2.5- and 1.5-fold) and CAM (3- and 1.6-fold) besides EAT cells accumulation in sub-G1 phase (20% and 18.38%), respectively. DISCUSSION AND CONCLUSION: Considering the potent anti-angiogenic and pro-apoptotic properties, lead molecules from EA extracts of E. jambolana and M. paradisiaca can be developed into anticancer drugs.

Synthesis and biological evaluation of novel isoxazolines linked via piperazine to 2- benzoisothiazoles as potent apoptotic agents.

Synthesis of 3-(4-((3-Phenyl-4,5-dihydroisoxazol-5-yl)methyl)piperazin-1-yl) benzoisothiazole derivatives (5a-i), which constitute a new class of isoxazolines, has been accomplished in regio-selective manner. These derivatives have been prepared by employing the reaction between substituted aldoximes (4a-i) and 3-(4-Allylpiperazin-1-yl) benzoisothiazole in presence of chloramine-T which afforded in good yields. These compounds were screened for cytotoxic activity on tumor cells. Four among the nine synthesized compounds were found to exhibit potent cytotoxic and antineoplastic activities in comparison to tumor necrosis factor-related apoptosis inducing ligand (TRAIL) protein in mammalian cancer cells. The rest of the derivatives showed moderate activity.

Combinatorial treatment with anacardic acid followed by TRAIL augments induction of apoptosis in TRAIL resistant cancer cells by the regulation of p53, MAPK and NFκß pathways.

TRAIL, an apoptosis inducing cytokine currently in phase II clinical trial, was investigated for its capability to induce apoptosis in six different human tumor cell lines out of which three cell lines showed resistance to TRAIL induced apoptosis. To investigate whether Anacardic acid (A1) an active component of Anacardium occidentale can sensitize the resistant cell lines to TRAIL induced apoptosis, we treated the resistant cells with suboptimal concentration of A1 and showed that it is a potent enhancer of TRAIL induced apoptosis which up-regulates the expression of both DR4 and DR5 receptors, which has been observed in the cellular, protein and mRNA levels. The death receptors upregulation consequent to A1 treatment was corroborated by the activation of p53 as well as phosphorylation of p38 and JNK MAP kinases and concomitant inactivation of NFκß and ERK signaling cascades. Also, A1 modulated the expression of key apoptotic players like Bax, Bcl-2 and CAD along with the abatement of tumor angiogenesis in vivo in EAT mouse model. Thus, post A1 treatment the TRAIL resistant cells turned into TRAIL sensitive cells. Hence our results demonstrate that A1 can synergize TRAIL induced apoptosis through the upregulation of death receptors and downregulation of anti-apoptotic proteins in cancer context.

P300 and neuropsychological measurements in patients with schizophrenia and their healthy biological siblings.

BACKGROUND: Patients with schizophrenia and their first-degree relatives' exhibit abnormalities in neuropsychological and electrophysiological measures especially P300. The aim was to study the P300 and neuropsychological measures together in patients with schizophrenia, their unaffected siblings, and normal controls. MATERIALS AND METHODS: Thirty patients diagnosed as schizophrenia, their unaffected biological siblings, and normal controls were included in the study. Inpatient group, the severity of symptoms was assessed by Positive and Negative Syndrome Scale. All subjects were administered P300 event-related potential, which was measured using oddball paradigm and specific neuropsychological tests from NIMHANS neuropsychiatry battery. RESULTS: Both patients with schizophrenia and their unaffected biological siblings showed lower P300 amplitude and longer P300 latency when compared with the normal controls. The three groups showed statistically significant differences in digit symbol substitution test, digit vigilance test, Trail making test B and Stroop test (P < 0.001). CONCLUSION: Patients with schizophrenia and their unaffected biological siblings show deficits in both cognitive function tests and P300 event-related potential. Our results suggest a continuum in the electrophysiological and neuropsychological measures among the three groups.

Anti-metastatic action of anacardic acid targets VEGF-induced signalling pathways in epithelial to mesenchymal transition.

Anacardic acid is a major constituent of nutshell of cashew. In this study, we have isolated it from the leaves of Anacardium Occidentale L. using polarity-based fractionation and confirmed the structure using GC-MS, NMR and FT-IR. The main focus of this study is to harness the molecular mechanism of anti-metastatic action of anacardic acid (A1). We have used MCF-7, a weak metastatic and U-87, a highly metastatic, breast and glioma cell lines respectively, for our study. We have shown that VEGF increases migration and invasion activities of MCF-7 cells, upon overexpression of Twist and Snail genes. It is observed from the current study that exposure of MCF-7 cells to A1 resulted in upregulation of epithelial marker E-cadherin with a concomitant decrease in the expression of mesenchymal markers Twist and Snail gene expression besides exhibiting a strong anti-migratory and anti-invasive activity. In metastatic U-87 glioma cells, treatment with A1 decreased the phosphorylation of MAP kinases, inhibited the translocation of Sp1 and down regulated VEGF and Flt-1 gene expression. Overall, the current findings demonstrate for the first time that anacardic acid functions as a potent EMT inhibitor by targeting VEGF signaling pathway, providing a novel template for drug discovery.

Crosstalk between VEGF and MTA1 signaling pathways contribute to aggressiveness of breast carcinoma.

The expression of metastasis associated protein (MTA1) correlates well with tumor metastasis; however its role as a proangiogenic protein and the molecular mechanisms underlying the same are not fully understood. In this study the MTA1 protein was expressed and purified to evaluate its angiogenic potential. In both MCF-7 and MDA-MB-231 cells, endogenous MTA1 protein was localized in the nucleus; while added recombinant MTA1 protein was bound to cell membrane as per immunofluorescence data. MTA1 was detected both in conditioned media and in human serum samples. Recombinant MTA1 regulated cellular functions of HUVEC's such as, proliferation, tube formation, and migration. MTA1 was more potent than VEGF in inducing invasion of breast cancer cells. Analogous to VEGF, MTA1 could induce angiogenesis in both non-tumor and tumor context, as verified by rat cornea, shell less CAM and xenograft models respectively. However MTA-1 was more potent an inducer of angiogenesis. VEGF or Flt-1 gene promoter, luciferase gene reporter analysis revealed that MTA1 up regulates the expression of VEGF and its receptor Flt-1 genes. Kinetics of VEGF-induced expression of MTA1 and qPCR studies showed that there is an increased expression of MTA1 in tumor cells. VEGF induced phosphorylation of endogenous MTA1 on tyrosine residues; phosphorylation was mediated through VEGFR2 and p38-MAP kinase. Recombinant MTA1 activated signaling, in MCF-7 and MDA-MB-231 cells, involved ERK and JNK pathways. In conclusion, MTA1 is a potent angiogenic molecule and cross talk between VEGF and MTA1 protein regulates tumor angiogenesis and metastasis.

Characteristics of the positive ion source at reduced gas feed.

The neutral beam injector of steady state superconducting tokamak (SST1-NBI) at IPR is designed for injecting upto 1.7 MW of neutral beam (Hº, 30-55 keV) power to the tokamak plasma for heating and current drive. Operations of the positive ion source (PINI or Plug-In-Neutral-Injector) of SST1-NBI were carried out on the NBI test stand. The PINI was operated at reduced gas feed rate of 2-3 Torr l/s, without using the high speed cryo pumps. Experiments were conducted to achieve a stable beam extraction by optimizing operational parameters namely, the arc current (120-300 A), acceleration voltage (16-40 kV), and a suitable control sequence. The beam divergence, power density profiles, and species fractions (H(+):H2(+):H3(+)) were measured by using the diagnostics such as thermal calorimetry, infrared thermography, and Doppler shift spectroscopy. The maximum extracted beam current was about 18 A. A further increase of beam current was found to be limited by the amount of gas feed rate to the ion source.

Mobilization of manganese by basalt associated Mn(II)-oxidizing bacteria from the Indian Ridge System.

The Indian Ridge System basalt bearing Mn-oxide coatings had todorokite as the major and birnesite as the minor mineral. We posit that microorganisms associated with these basalts participate in the oxidation of Mn and contribute to mineral deposition. We also hypothesized that, the Mn-oxidizing microbes may respond reversibly to pulses of fresh organic carbon introduced into the water column by mobilizing the Mn in Mn-oxides. To test these two hypotheses, we enumerated the number of Mn-oxidizers and -reducers and carried out studies on the mobilization of Mn by microbial communities associated with basalt. In medium containing 100 µM Mn(2+), 10(3) colony forming units (CFU) were recovered with undetectable number of reducers on Mn-oxide amended medium, suggesting that the community was more oxidative. Experiments were then conducted with basalt fragments at 4±2 °C in the presence 'G(+)' and absence 'G(-)' of glucose (0.1%). Controls included set-ups, some of which were poisoned with 15 mM azide and the others of which were heat-killed. The mobilization of Mn in the presence of glucose was 1.76 µg g(-1) d(-1) and in the absence, it was 0.17 µg g(-1) d(-1) after 150 d. Mn mobilization with and without added glucose was 13 and 4 times greater than the corresponding azide treated controls. However, rates in 'G(+)' were 16 times and 'G(-)' 24 times more than the respective heat killed controls. The corresponding total counts in the presence of added glucose increased from 1.63×10(6) to 6.71×10(7) cells g(-1) and from 1.41×10(7) to 3.52×10(7) cells g(-1) in its absence. Thus, the addition of glucose as a proxy for organic carbon changed the community's response from Mn(II)-oxidizing to Mn(IV)-reducing activity. The results confirm the participation of Mn oxidizing bacteria in the mobilization of Mn. Identification of culturable bacteria by 16S rRNA gene analysis showed taxonomic affiliations to Bacillus, Exiguobacterium, Staphylococcus, Brevibacterium and Alcanivorax sp.

Buffalo colostrum ß-lactoglobulin inhibits VEGF-induced angiogenesis by interacting with G protein-coupled receptor kinase.

ß-lactoglobulin (ß-lg), a major whey protein was purified and characterised from buffalo colostrum. The in silico analysis of the tryptic peptides based on LC-CID-MS/MS facilitated the identification of protein as ß-lg. The sequences IIVTQ f[1-5] and LSFNPTQLEEQCHV f(149-162) of m/z 933(+) and 851(2+) were found to match N- and C-extreme of ß-lg while IDALNENK f(84-91) and TPEVDDEALEKFDK f(125-138) sequences deduced for m/z 916(+) and 818(2+) were in compliance to buffalo milk ß-lg. Considering the sequence similarity of ß-lg to glycodelin, a proven angiogenic protein, similar role for ß-lg from buffalo colostrum (BLG-col) was examined. Interestingly, BLG-col exhibited anti-angiogenic activity by potently inhibiting cell proliferation, micro-vessel sprouting, cell migration and tube formation of human umbilical vein endothelial cells (HUVECs) in a dose-dependent manner but having varied effect on Ehrlich ascites tumor cells, MCF-7, MDA-MB 435 and MDA-MB 231 cell lines. The anti-angiogenic potential of BLG-col was found to be vascular endothelial growth factor mediated. The immunolocalisation of BLG-col on the cell surface of HUVECs evidenced using FITC-labelled ß-lg antibody indicated its extra-cellular binding. Furthermore, BLG-col interacting HUVEC membrane protein (64 kDa) was detected by immunoblot and its identity was established by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry analysis, which showed peptide sequence homology to G protein-coupled receptor kinase 4.

Crosstalk between VEGF and novel angiogenic protein regulates tumor angiogenesis and contributes to aggressiveness of breast carcinoma.

We have identified and characterized a novel proangiogenic glycoprotein (NAP) with molecular weight of 67 kDa from synovial fluid of rheumatoid arthritis patients. Proteomic analysis of the protein revealed 29% sequence coverage with maximum identity for human retinoblastoma binding protein 2. N-terminal amino acid sequence showed no identity to recently discovered protein sequences. NAP was also identified in both normal and tumor cell lines by Western blotting. NAP is a permeability factor as verified by miles permeability assay. The proangiogenic potential of NAP was identified using shell less CAM, rat cornea and tumor on CAM assays. NAP induces expression of VEGF and Flt-1 gene as verified by promoter reporter gene analysis. Further NAP induces proliferation of endothelial cells and formation of tube like structures. NAP is also involved in migration and invasion of tumor cells. Clinical data revealed the presence of NAP in breast cancer biopsies. We have developed monoclonal antibody (mAb), and specific ELISA, which confirmed the presence of NAP in the cytosol of tumor cells. The mAb effect was evaluated with established angiogenic assays. Further, we investigated the detailed mechanism by which NAP induces angiogenesis. NAP is phosphorylated by VEGF induced activation of MAPK and JNK pathways through VEGFR2 phosphorylation. NAP involves JNK pathway predominantly with further activation of NFκB in downstream processing of VEGF activation. Together these findings establish that NAP displays angiogenic properties and promotes efficient neovascularization both in vitro and in vivo models. These observations suggest that anti-NAP-mAb can be targeted for antiangiogenic therapy of cancer.