Pisolithus indicus, a new species of ectomycorrhizal fungus associated with Dipterocarps [corrected] in India

Pisolithus is cosmopolitan in both tropical and temperate regions and forms ectomycorrhizal associations with a wide range of woody plants. Pisolithus indicus, a new species associated with Vateria indica (Dipterocarpaceae) is reported in this study from a dipterocarp native forest in the Western Ghats in India, using both morphological and molecular tools. The length of ITS1 and ITS2 regions of the present collection differed with other sequences of Pisolithus available in the databases. Phylogenetic analysis indicates that this species did not show significant homology with existing Pisolithus sequences reported previously and formed a separate branch linking with another Pisolithus isolate from dipterocarps. Molecular and morphological evidence showed that P. indicus is a new species associated with dipterocarps in India.     

Molecular characterization of calmodulin trapping by calcium/calmodulin-dependent protein kinase II

Autophosphorylation of alpha-Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) at Thr(286) results in calmodulin (CaM) trapping, a >10,000-fold decrease in the dissociation rate of CaM from the enzyme. Here we present the first site-directed mutagenesis study on the dissociation of the high affinity complex between CaM and full-length CaM kinase II. We measured dissociation kinetics of CaM and CaM kinase II proteins by using a fluorescently modified CaM that is sensitive to binding to target proteins. In low [Ca(2+)], the phosphorylated mutant kinase F293A and the CaM mutant E120A/M124A exhibited deficient trapping compared with wild-type. In high [Ca(2+)], the CaM mutations E120A, M124A, and E120A/M124A and the CaM kinase II mutations F293A, F293E, N294A, N294P, and R297E increased dissociation rate constants by factors ranging from 2.3 to 116. We have also identified residues in CaM and CaM kinase II that interact in the trapped state by mutant cycle-based analysis, which suggests that interactions between Phe(293) in the kinase and Glu(120) and Met(124) in CaM specifically stabilize the trapped CaM-CaM kinase II complex. Our studies further show that Phe(293) and Asn(294) in CaM kinase II play dual roles, because they likely destabilize the low affinity state of CaM complexed to unphosphorylated kinase but stabilize the trapped state of CaM bound to phosphorylated kinase.     

Modulation of Carbohydrate Metabolism During <Emphasis Type="Italic">N</Emphasis>-Methyl <Emphasis Type="Italic">N</Emphasis>-Nitrosourea Induced Neurotoxicity in Mice: Role of Curcumin

The present study was carried out to elucidate the effectiveness of curcumin in mitigating the adverse effects caused by N-Methyl N-Nitrosourea (MNU) on mouse cerebellum and cerebrum. Male laca mice received either intravenous MNU treatment at a dose of 10 mg/kg bw in sterile double distilled water, curcumin alone 60 mg/kg bw in drinking water, or combined MNU and curcumin treatment on alternate days for a period of 2 months. The effects of different treatments were studied on carbohydrate metabolizing enzymes viz: hexokinase, glucose-6-phosphatase (G6P), glucose-6-isomerase (G6I), lactate dehydrogenase (LDH), succinate dehydrogenase (SDH) and glycogen levels. Curcumin supplementation to MNU treated mice was able to reduce significantly the activities of the G6P, G6I, hexokinase, LDH, SDH and increased the glycogen contents in both the regions of brain which were altered following MNU treatment. Hence, curcumin shall prove to be effective in ameliorating the adverse effects caused by MNU.     

Nifedipine loaded chitosan microspheres prepared by emulsification phase-separation

A high yield of nifedipine-chitosan microspheres could be obtained using an emulsification phase-separation method. A high level of entrapment of nifedipine in the microspheres was achieved. The microspheres exhibited excellent swelling properties. Differential scanning calorimetry, X-ray diffractometry, and scanning electron microscopy confirmed that at 1.84% loading, nifedipine was dispersed molecularly. The microspheres exhibited faster release at low loadings compared to high loadings. Fitting the data to the coupled Fickian/case II equation, showed that at low loadings polymer relaxation coefficients (k₂) were high. As the polymer content increased in the microspheres, the value of n (diffusional exponent characteristic of the release mechanism) approached one, which is indicative of zero order.     

Calcitonin receptor-mediated CFTR activation in human intestinal epithelial cells

High levels of calcitonin (CT) observed in medullary thyroid carcinoma and other CT‐secreting tumours cause severe diarrhoea. Previous studies have suggested that CT induces active chloride secretion. However, the involvement of CT receptor (CTR) and the molecular mechanisms underlying the modulation of intestinal electrolyte secreting intestinal epithelial cells have not been investigated. Therefore, current studies were undertaken to investigate the direct effects of CT on ion transport in intestinal epithelial cells. Real time quantitative RT‐PCR and Western blot analysis demonstrated the expression of CTR in intestinal epithelial T84 cells. Exposure of T84 cells to CT from the basolateral but not from apical side significantly increased short circuit current (I_SC) in a dose‐dependent manner that was blocked by 1 μM of CTR antagonist, CT8–32. CT‐induced I_SC was blocked by replacing chloride in the bath solutions with equimolar gluconate and was significantly inhibited by the specific cystic fibrosis transmembrane conductance regulator (CFTR) inhibitor, CFTR_127inh. Further, biotinylation studies showed that CT increased CFTR levels on the apical membrane. The presence of either the Ca²⁺ chelator, bis(2‐aminophenoxy)ethane tetraacetic acid‐acetoxymethyl (BAPTA‐AM) ester or the protein kinase A (PKA) inhibitor, H89, significantly inhibited I_SC induced by CT (～32–58% reduction). Response to CT was retained after permeabilization of the basolateral or the apical membranes of T84 cells with nystatin. In conclusion, the activation of CTR by CT induced chloride secretion across T84 monolayers via CFTR channel and the involvement of PKA‐ and Ca²⁺‐dependent signalling pathways. These data elucidate the molecular mechanisms underlying CT‐induced diarrhoea.     

Serum levels of angiogenic and anti-angiogenic factors: their prognostic relevance in locally advanced hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is a prototype tumor wherein angiogenesis plays a vital role in its progression. The role of VEGF, a major angiogenic factor in HCC is known; however, the role of anti-angiogenic factors simultaneously with the angiogenic factors has not been studied before. Hence, in this study, the serum levels of major angiogenic [Vascular Endothelial Growth Factor (VEGF), angiopoietin-2 (Ang-2)] and anti-angiogenic (endostatin, angiostatin) factors were analyzed and correlated with clinico-radiological features and with outcome. A total of 150 patients (50 HCC, 50 cirrhosis and 50 chronic hepatitis) and 50 healthy controls were enrolled in this study. Serum levels of VEGF, Ang-2, endostatin, and angiostatin were estimated by enzyme-linked immunosorbent assay. HCC shows significantly elevated serum levels of angiogenic factors VEGF and Ang-2 and of anti-angiogenic factors endostatin and angiostatin. ROC curve analysis for serum VEGF yielded an optimal cut-off value of 225.14 pg/ml, with a sensitivity of 78 % and specificity of 84.7 % for a diagnosis of HCC and its distinction from other group. Using this value, the univariate and multivariate analysis revealed significantly poor outcome in patients with higher levels of serum VEGF (p = 0.009). Combinatorial analysis revealed that patients with higher levels of both angiogenic and anti-angiogenic factors showed poor outcome. Serum VEGF correlates with poor survival of HCC patients and, therefore, serves as a non-invasive biomarker of poor prognosis. Moreover, elevated levels of anti-angiogenic factors occur endogenously in HCC patients.     

Phylogenetic position of Indian termites (Isoptera: Termitidae) with their respective genera inferred from DNA sequence analysis of the mitochondrial cytochrome oxidase gene subunit I compared to subunit II

It has been shown that over-expression of Special AT-rich binding protein 1 (SATB1) in breast cancer predicts a poor prognosis. This study was aimed at investigating the effects of silencing SATB1 on mesenchymal derived human osteosarcoma U2OS cells and the underlying mechanisms. The expressions of SATB1 and the related genes in the cells were detected by qRT-PCR and/or Western Blotting. SATB1 silencing was achieved by stable transfection with the vectors expressing small hairpin RNA versus SATB1. Cell proliferation was detected in a microplate reader with Cell Counting Kit-8 and the cell cycle was analyzed by flow cytometry using a cell cycle detection kit. The study found that SATB1 was particularly over-expressed in human osteosarcoma U2OS. Silencing SATB1 inhibited the proliferation of U2OS. It was found that inhibition of cell proliferation resulted from cell cycle arrest due to down-regulated expression of CFGF and JunB. The over-expression of SATB1 is responsible for abnormal proliferation of mesenchymal derived human Osteosatcoma U2OS cells, indicating that silencing SATB1 expression in the cells might be developed as an efficient osteosarcoma therapy. CTGF and JunB were involved in SATB1-mediated proliferation of U2OS cells.     

Integrated sedimentological,ichnological and sequence stratigraphical studies of the Koti Dhaman Formation (Tal Group), Nigali Dhar Syncline,Lesser Himalaya,India: paleoenvironmental,paleoecological, paleogeographic significance

Abstract

Integrated ichnology, sedimentology and sequence stratigraphy of the Lower Quartzite Member to the Arkosic Sandstone Member of the Koti Dhaman Formation (Cambrian Series 2, Stage 4), Tal Group, Nigali Dhar Syncline, Lesser Himalayan lithotectonic zone are presented. Trilobite traces of Gondwanan affinity i.e., Cruziana salomonis, Cruziana fasciculata, Rusophycus dispar and Rusophycus burjensis are recorded along with Arenicolites isp. and Skolithos isp. from the Lower Quartzite Member. A rich and diverse ichnoassemblage attributed to the Cruziana ichnofacies is described for the first time from the Arkosic Sandstone Member of the same formation. Seven ichnofossil assemblages, i.e., Cruziana-Rusophycus, Planolites-Palaeophycus, Cruziana problematica, Diplichnites, Cochlichnus anguineus, Bergaueria perata and Psammichnites gigas have been recognized in the Lower Quartzite to Arkosic Sandstone members of the Koti Dhaman Formation. Seven sedimentary facies i.e., sandstone–shale facies (FT1), cross-bedded (trough and planar) sandstone (FT2), bedded sandstone facies (FT3), shale facies (FT4), shale–sandstone facies (FT5), shale-rippled sandstone facies (FT6) and planar and trough cross-laminated sandstone (FT7) and four facies associations FA1-FA4 are identified in the Koti Dhaman Formation. The formation contains shallowing upward parasequences of a tidal flat complex. Overall, two major events are recognized: i) the break in sedimentation between the Lower Quartzite Member and the overlying Shale Member probably related to forced-regressive event and ii) the facies shift from FT6 to FT7 of the Arkosic Sandstone Member represents an erosive transgressive event; the surface is interpreted as wave ravinement surface, which also serves as a sequence boundary. Integrated ichnology, sedimentology and sequence stratigraphic studies indicate that the Lower Quartzite Member was deposited in a shallow subtidal sand sheet complex and tidal flat complex; the Shale Member was deposited in a mud flat setting of a tidal flat complex, and the Arkosic Sandstone Member in a mixed-flat (tidal flat complex) to sand sheet complex front and margin (subtidal sand sheet complex). Overall, the lower to middle part of the Koti Dhaman Formation represents a tide-dominated shallow subtidal–intertidal to mud-flat subenvironments of the tidal flat complex. A palaeogeographic reconstruction of lower Cambrian (516–514?Ma) is presented based on the distribution of trilobite traces from the Lesser Himalaya and the Bikaner–Nagaur area of Peninsular India (eastern Gondwana), Egypt, Jordan, Turkey (western Gondwana) and Canada (Avalonia).