Characterization of carbon fluxes, stock and nutrients in the sacred forest groves and invasive vegetation stands within the human dominated landscapes of a tropical semi-arid region.

In the semi-arid plains of Southern India, outside the protected area network, sacred groves forests and the barren lands invaded by Prosopis juliflora are reckoned to be the major greenery, but have homogenous and heterogeneous vegetation respectively. This study attempted to compare 50 Sacred Groves Stands (SGS) and 50 monodominant Prosopis juliflora Stands (PJS) for the functional diversity, evenness, floral diversity, carbon stock and dynamics, carbon-fixing traits, dendrochronology of trees, soil nutrient profiles, and soil erosion. Quadrat sample survey was adopted to record stand density, species richness, abundance, basal area and leaf area index; composite soil samples were collected at depths 0-30 cm for nutrient profiling (N, P, K, and OC). Photosynthesis rate (µmole co2 m2/sec), air temperature (°c), leaf intracellular co2 concentration (ppm), ambient photosynthetic active radiation (µmole m2/sec), transpiration rate (m. mole H2O m2/sec) were determined for the 51 tree species existed in SGS and PJS using Plant Photosynthesis system. Structural Equation Model (SEM) was applied to derive the carbon sequestering potential and photosynthetic efficiency of eight dominant tree species using vital input parameters, including eco-physiological, morphological, and biochemical characterization. The Revised Universal Soil Loss Equation (RUSLE) model, in conjunction with ArcGIS Pro and ArcGIS 10.3, was adopted to map soil loss. Carbon source/sink determinations inferred through Net Ecosystem Productivity (NEP) assessments showed that mature SGS potentially acted as a carbon sink (0.06 ± 0.01 g C/m2/day), while matured PJS acted as a carbon source (-0.34 ± 0.12 g C/m2/day). Soil erosion rates were significantly greater (29.5 ± 13.4 ton/ha/year) in SGS compared to PJS (7.52 ± 2.55 ton/ha/year). Of the eight selected tree species, SEM revealed that trees belonging to the family Fabaceae [Wrightia tinctoria (estimated coefficient: 1.28, p = 0.02) > Prosopis juliflora (1.22, p = 0.01) > Acacia nilotica (1.21, p = 0.03) > Albizia lebbeck (0.97, p = 0.01)] showed comparatively high carbon sequestering ability.

An economic evaluation of the randomized controlled trial of topical corticosteroid and home-based narrowband ultraviolet B for active and limited vitiligo (the HI-Light Vitiligo Trial).

BACKGROUND: Economic evidence for vitiligo treatments is absent. OBJECTIVES: To determine the cost-effectiveness of (i) handheld narrowband ultraviolet B (NB-UVB) and (ii) a combination of topical corticosteroid (TCS) and NB-UVB compared with TCS alone for localized vitiligo. METHODS: Cost-effectiveness analysis alongside a pragmatic, three-arm, placebo-controlled randomized controlled trial with 9 months' treatment. In total 517 adults and children (aged ≥ 5 years) with active vitiligo affecting < 10% of skin were recruited from secondary care and the community and were randomized 1: 1: 1 to receive TCS, NB-UVB or both. Cost per successful treatment (measured on the Vitiligo Noticeability Scale) was estimated. Secondary cost-utility analyses measured quality-adjusted life-years using the EuroQol 5 Dimensions 5 Levels for those aged ≥ 11 years and the Child Health Utility 9D for those aged 5 to < 18 years. The trial was registered with number ISRCTN17160087 on 8 January 2015. RESULTS: The mean ± SD cost per participant was £775 ± 83·7 for NB-UVB, £813 ± 111.4 for combination treatment and £600 ± 96·2 for TCS. In analyses adjusted for age and target patch location, the incremental difference in cost for combination treatment compared with TCS was £211 (95% confidence interval 188-235), corresponding to a risk difference of 10·9% (number needed to treat = 9). The incremental cost was £1932 per successful treatment. The incremental difference in cost for NB-UVB compared with TCS was £173 (95% confidence interval 151-196), with a risk difference of 5·2% (number needed to treat = 19). The incremental cost was £3336 per successful treatment. CONCLUSIONS: Combination treatment, compared with TCS alone, has a lower incremental cost per additional successful treatment than NB-UVB only. Combination treatment would be considered cost-effective if decision makers are willing to pay £1932 per additional treatment success.

Randomized controlled trial of topical corticosteroid and home-based narrowband ultraviolet B for active and limited vitiligo: results of the HI-Light Vitiligo Trial.

BACKGROUND: Evidence for the effectiveness of vitiligo treatments is limited. OBJECTIVES: To determine the effectiveness of (i) handheld narrowband UVB (NB-UVB) and (ii) a combination of potent topical corticosteroid (TCS) and NB-UVB, compared with TCS alone, for localized vitiligo. METHODS: A pragmatic, three-arm, placebo-controlled randomized controlled trial (9-month treatment, 12-month follow-up). Adults and children, recruited from secondary care and the community, aged ≥ 5 years and with active vitiligo affecting < 10% of skin, were randomized 1 : 1 : 1 to receive TCS (mometasone furoate 0·1% ointment + dummy NB-UVB), NB-UVB (NB-UVB + placebo TCS) or a combination (TCS + NB-UVB). TCS was applied once daily on alternating weeks; NB-UVB was administered on alternate days in escalating doses, adjusted for erythema. The primary outcome was treatment success at 9 months at a target patch assessed using the participant-reported Vitiligo Noticeability Scale, with multiple imputation for missing data. The trial was registered with number ISRCTN17160087 on 8 January 2015. RESULTS: In total 517 participants were randomized to TCS (n = 173), NB-UVB (n = 169) and combination (n = 175). Primary outcome data were available for 370 (72%) participants. The proportions with target patch treatment success were 17% (TCS), 22% (NB-UVB) and 27% (combination). Combination treatment was superior to TCS: adjusted between-group difference 10·9% (95% confidence interval 1·0%-20·9%; P = 0·032; number needed to treat = 10). NB-UVB alone was not superior to TCS: adjusted between-group difference 5·2% (95% CI - 4·4% to 14·9%; P = 0·29; number needed to treat = 19). Participants using interventions with ≥ 75% expected adherence were more likely to achieve treatment success, but the effects were lost once treatment stopped. Localized grade 3 or 4 erythema was reported in 62 (12%) participants (including three with dummy light). Skin thinning was reported in 13 (2·5%) participants (including one with placebo ointment). CONCLUSIONS: Combination treatment with home-based handheld NB-UVB plus TCS is likely to be superior to TCS alone for treatment of localized vitiligo. Combination treatment was relatively safe and well tolerated but was successful in only around one-quarter of participants.

Point-of-care lung ultrasound in the assessment of suspected COVID-19: a retrospective service evaluation with a severity score.

INTRODUCTION: Point-of-care lung ultrasound (POCUS) has been advocated as a tool to assess the severity of COVID19 and thereby aid risk stratification. METHODS: We conducted a retrospective service evaluation between the 3rd March and the 5th May 2020 to describe and characterise the use of POCUS within an acute care pathway designed specifically for the assessment of suspected or confirmed COVID-19. A novel POCUS severity scale was formulated by assessing pleural and interstitial abnormalities within six anatomical zones (three for each lung). An aggregated score was calculated for each patient and evaluated as a marker of disease severity using standard metrics of discriminatory performance. RESULTS: POCUS was performed in the assessment of 100 patients presenting with suspected COVID-19. POCUS was consistent with COVID-19 infection in 92% (n = 92) of the patients assessed. Severity, as assessed by POCUS, showed good discriminatory performance to predict all-cause inpatient mortality, death or critical care admission, and escalated oxygen requirements (AUC .80, .80, 82). The risk of all-cause mortality in patients with scores in lowest quartile was 2.5% (95%CI 0.12- 12.95) compared with 42.9% (95CI 15.8 - 75.0%) in the highest quartile. POCUS assessed severity correlated with length of stay and duration of supplemental oxygen therapy. CONCLUSION: A simple aggregated score formed by the summating the degree of pleural and interstitial change within six anatomical lung zones showed good discriminatory performance in predicting a range of adverse outcomes in patients with suspected COVID-19.

Physiologically Based Pharmacokinetic Modeling of Fluorescently Labeled Block Copolymer Nanoparticles for Controlled Drug Delivery in Leukemia Therapy.

A physiologically based pharmacokinetic (PBPK) model was developed that describes the concentration and biodistribution of fluorescently labeled nanoparticles in mice used for the controlled delivery of dexamethasone in acute lymphoblastic leukemia (ALL) therapy. The simulated data showed initial spikes in nanoparticle concentration in the liver, spleen, and kidneys, whereas concentration in plasma decreased rapidly. These simulation results were consistent with previously published in vivo data. At shorter time scales, the simulated data predicted decrease of nanoparticles from plasma with concomitant increase in the liver, spleen, and kidneys before decaying at longer timepoints. Interestingly, the simulated data predicted an unaccounted accumulation of about 50% of the injected dose of nanoparticles. Incorporation of an additional compartment into the model justified the presence of unaccounted nanoparticles in this compartment. Our results suggest that the proposed PBPK model can be an excellent tool for prediction of optimal dose of nanoparticle-encapsulated drugs for cancer treatment.

Hepatoprotection by active fractions from Desmostachya bipinnata stapf (L.) against tamoxifen-induced hepatotoxicity.

OBJECTIVE: The aim was to evaluate the effect of the polyphenolic fraction of Desmostachya bipinnata Stapf (PFDB) (Poaceae) on tamoxifen (TAM)-induced liver damage in female Sprague-Dawley rats. MATERIALS AND METHODS: The roots of Desmostachya bipinnata were extracted in 70% methanol, and the polyphenolic fraction was isolated. Protection of BRL3A cells against ethanol-induced damage was determined by 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay. Hepatotoxicity was induced in rats by oral administration of TAM (45 mg/kg/day) for 21 days. The PFDB was administered to experimental animals at two selected doses (100 and 200 mg/kg/day) during the treatment. The serum levels of various biochemical parameters and the antioxidant enzymes were examined by standard procedures. RESULTS: A dose-dependent increase in percentage viability was observed when ethanol-exposed BRL3A cells were treated with PFDB. Both the treatment groups upon pretreatment with PFDB exhibited a significant (P ≤ 0.05) protective effect by lowering serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase, triglycerides, cholesterol, urea, uric acid, bilirubin and creatinin levels and improving protein level in serum in dose-dependent manner, which was comparable to that of silymarin group. In addition, PFDB prevented elevation of reduced glutathione, glutathione peroxidase, superoxide dismutase and catalase in the TAM-intoxicated rats in concentration-dependent manner and significantly (P < 0.05) reduced the lipid peroxidation in the liver tissue. The biochemical observations were supplemented with histopathological reports, which showed the attenuation of hepatocellular necrosis. CONCLUSIONS: The results of this study strongly indicate that the polyphenolic fraction of the plant roots has a potent hepatoprotective action against TAM-induced hepatic damage in rats.

Protective effect of Monascus fermented rice against STZ-induced diabetic oxidative stress in kidney of rats.

In the present study, anti-diabetic activity and nephroprotective effect of MMFR was evaluated by using STZ-induced diabetic rats. Administration of MMFR at 100 and 200 mg/kg bw showed significant (P < 0.01) anti-hyperglycemic activity by lowering blood glucose level, HbA1C and increasing body weight. Altered lipid profiles in diabetic rats were restored to normal level on treatment with MMFR and showed significant (P < 0.01) decrease in the elevated levels of biochemical parameters. MMFR produced significant (P < 0.01) improvement in antioxidant levels in kidney. Food safety of MFR produced by using mutant Monascus purpureus 254 (MMFR) was evaluated for genotoxicity and oral acute toxicity. In Ames mutagenicity assay MMFR doesn't showed any toxicity to the test strain S. typhimurium till 5 mg/plate. Acute toxicity study also recorded no toxicity till the dose of 5,000 mg/kg bw. The study concluded that MMFR, not only possess anti-diabetic activity but also prevents nephropathy and hypercholesterolemia due to diabetes.

Clinical nanomedicine: a solution to the chemotherapy conundrum in pediatric leukemia therapy.

Although chemotherapy-based treatment of leukemia has tremendously improved survival rates in children, induction of treatment-related side effects is a major concern in clinical oncology. The development of nanotechnology-based drug delivery techniques to target clinically approved anticancer agents specifically to leukemic cells should diminish toxic side effects. This review aims to address the rational design of nanotherapeutics in treating hematologic malignancies with a focus on acute lymphoblastic leukemia (ALL)--the most prominent form of pediatric cancer.

Synthesis, in-vitro antimicrobial and antitubercular screening of Schiff bases of 3-amino-1-phenyl-4- [2-(4-phenyl-1,3-thiazol-2-yl) hydrazin-1-ylidene]-4,5-dihydro-1H-pyrazol-5-one.

PURPOSE: Synthesis and antimicrobial activity of some Schiff bases of 3-amino-1-phenyl-4- [2-(4-phenyl-1,3-thiazol-2-yl) hydrazin-1-ylidene]-4,5-dihydro-1H-pyrazol-5-ones (TZP4a-l) are described. MATERIALS AND METHODS: Structures of the synthesized compounds were confirmed using infrared, (1)H nuclear magnetic resonance, and mass spectral data. Synthesized compounds were tested in-vitro against four Gram-positive and four Gram-negative bacterial strains, three fungal strains and two mycobacterial strains. Title compounds were screened its in-vitro cytotoxicity (IC50) by 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay using mouse embryonic fibroblasts cell line (NIH 3T3). RESULTS AND DISCUSSION: Compounds TZP4 g and TZP4 h were found to be significant activity against Bacillus substilis (bacteria) and Aspergillus niger (fungi). In-vitro anti-tuberculosis (TB) activity of compound TZP4g showed appreciable antitubercular activity against Mycobacterium tuberculosis H37Rv strain (minimum inhibitory concentration [MIC] =0.6.48 × 10(-3) µM/mL) which was 1.69 and 3.91 times more active than the standard drug, pyrazinamide (25.38 × 10(-3) µM/mL) and streptomycin (MIC = 11.01 × 10(-3) µM/mL), respectively. Their in-vitro cytotoxicity (IC50) was determined to establish a selectivity index (SI) (SI = IC50/MIC). Compounds TZP4 c, TZP4 g, and TZP4 h have SI 82.85, 168.88, and 199.07, respectively. CONCLUSION: All the title compounds had mild toxicity on the mouse embryonic fibroblasts NIH 3T3 cells (IC50 ≥ 100 µM). In comparison to the results of toxicity and antimycobacterial activity tests, it was observed that the activity of the compounds is not due to general toxicity effect; however, their antimycobacterial activity can be possibly because of their selective antimycobacterial effect. We concluded from our investigations that TZP4 c, TZP4 g, and TZP4 h may be considered promising for the development of new anti-TB agents.

Synthesis of some new thioxoquinazolinone derivatives and a study on their anticonvulsant and antimicrobial activities.

OBJECTIVE: A series of ten novel derivatives of 3-substituted-2-thioxoquinazolin-4(3H)-ones have been synthesized from anthranilic acid via Mannich reaction with various secondary amines in presence of formaldehyde in ice cold condition. MATERIALS AND METHODS: The structure of these compounds have been elucidated by spectral (FTIR, 1H-NMR and mass) analysis. The titled compounds were evaluated for antimicrobial and anticonvulsant activities. Antimicrobial activities were determined by cup plate method and MIC values using the micro dilution broth method against two Gram positive bacteria Staphylococcus aureus and Streptococcus aureus, two Gram negative bacteria Escherichia coli and Proteus vulgaris and against two fungi Candida albicans and Aspergillus niger. Amikacin and fluconazole were used as standard antibacterial and antifungal agents in the concentration of 10 µg/disc 20 µg/disc respectively. RESULTS AND CONCLUSIONS: Amongst the compounds tested, compound 2-(2,3-dimethylphenyl) (3-(4-ethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydroquinazolin-1-2H)-1ylmethyl amino)benzoic acid (PTQ-03) and 2-((2,3-dimethylphenyl)((3-(4-ethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydroquinazolin-1(2H)-yl)methyl)amino)benzoic acid (ETQ-03) showed broad spectrum of activity against all the tested Gram positive bacteria, Gram negative bacteria and the fungi. Anti-convulsant activity of the compounds was evaluated by maximal electro shock (MES) convulsion method. The compounds sodium 2-(2-((2,6-Dichlorophenyl)(3-(4-oxo-2-thioxo-3,4-dihydroquinazolin-1(2H)-yl)methyl)amino) phenyl acetate (PTQ-04) and N-(4-Hydroxyphenyl)-N-((3-naphthalen-2-yl)-4-oxo-2-thioxo-3,4-dihydorquinazolin-1(2H)-ylmethyl)acetamide (NTQ-01) showed potent anticonvulsant activity.

Designer foods and their benefits: A review.

Designer foods are normal foods fortified with health promoting ingredients. These foods are similar in appearance to normal foods and are consumed regularly as a part of diet. In this article we have reviewed the global regulatory status and benefits of available designer foods such as designer egg, designer milk, designer grains, probiotics, designer foods enriched with micro and macronutrients and designer proteins. Designer foods are produced by the process of fortification or nutrification. With the advances in the biotechnology, biofortification of foods using technologies such as recombinant DNA technology and fermentation procedures are gaining advantage in the industry. The ultimate acceptability and extensive use of designer foods depend on proper regulation in the market by the regulatory authorities of the country and by creating consumer awareness about their health benefits through various nationwide programs.

Tuberculosis is still a scourge of mankind--why?

This article focuses on issues and challenges regarding why tuberculosis (TB) remains as horrible scourge to mankind even today that too in the era of nanotechnology. It tries to evoke a uniform consciousness among treating doctorsboth at government and private levels to follow a uniform diagnostic algorithm and treatment protocols which is very much readily available under Revised National Tuberculosis Control Programme (RNTCP) Directly Observed Treatment Short course (DOTS) throughout our nation. Even today TB is the singlemost major infectious killer disease which claims 2 deaths in every 3 minutes in India which has one-fifth of global TB burden. The challenges faced are highlighted here like issues of uniformity in treatment, multidrug resistant TB (MDR TB), extensive drug resistant TB (XDR TB), HIV-TB coinfection, diabetes and TB, etc. In spite of nationwide coverage of RNTCP DOTS, every practitioner is not strictly or willfully following the guidelines prescribed. If the guidelines are maintained one hundred per cent with full involvement of Indian Medical Association/Indian Academy of Pediatrics and various other medical organisations then the author can confidently feels that the day is not far to combat and contain the horrible scourge. Time has come that every medical practitioner should take an oath to evolve a uniform consciousness to adhere to the guidelines prescribed under RNTCP DOTS which may prevent the future generation to succumb to the horrible scourge of TB. Shall we?

Simultaneous RP-HPLC method for the stress degradation studies of atorvastatin calcium and ezetimibe in multicomponent dosage form

A stability-indicating reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for simultaneous estimation of atorvastatin calcium and ezetimibe for their multicomponent dosage form. The proposed RP-HPLC method utilizes a 125 mm x 4.6 mm i.d 5 ¦Ìm Phenomenex C-18 column at ambient temperature; the optimum mobile phase consists of acetonitrile and 0.4% v/v triethylamine (pH adjusted to 5.5 with ortho-phosphoric acid) in the ratio of 55:45, v/v respectively, flow rate of 1.0 ml/min. Measurements were made at a wavelength of 231 nm. Multicomponent dosage form was exposed to thermal, photolytic, hydrolytic and oxidative stress. No co eluting, interfering peaks from excipients, impurities were observed for the degradation products and hence the method was found to be specific. The method was linear in the range of 5-25 ¦Ìg/ml for atorvastatin calcium and ezetimibe. The mean recoveries were 98.82% and 98.72% for atorvastatin calcium and ezetimibe respectively. The method was validated for linearity, range, precision, accuracy, specificity, selectivity, intermediate precision, ruggedness, robustness, solution stability and suitability(AU)

Se desarroll¨® y valid¨® un m¨¦todo estable de cromatograf¨ªa l¨ªquida de alta eficacia de fase reversa (RP-HPLC) para la estimaci¨®n simult¨¢nea de atorvastatina de calcio y ezetimiba en su forma de dosificaci¨®n multicomponente. El m¨¦todo RP-HPLC propuesto utiliza, a temperatura ambiente, una columna C-18 Phenomenex de 125 mm x 4,6 mm y d.i de 5 ¦Ìm; la fase m¨®vil ¨®ptima consta de acetonitrilo y 0,4% v/v de trietilamina (pH ajustado a 5,5 con ¨¢cido ortofosf¨®rico) en una proporci¨®n de 55:45, v/v, respectivamente, y una velocidad de flujo de 1,0 ml/min. Las medidas se realizaron a una longitud de onda de 231 nm. La forma de dosificaci¨®n multicomponente se expuso a estr¨¦s oxidativo, hidrol¨ªtico, fotol¨ªtico y t¨¦rmico. No se observaron, en la degradaci¨®n de productos, ni impurezas ni picos de coeluci¨®n o interferencia por excipientes, y, adem¨¢s, el m¨¦todo result¨® ser espec¨ªfico. El m¨¦todo fue linear, en el rango de 5-25 ¦Ìg/ml para atorvastatina de calcio y ezetimiba. Las recuperaciones medias fueron del 98,82% y 98,72% para atorvastatina de calcio y ezetimiba, respectivamente. El m¨¦todo se valid¨® para linealidad, rango, precisi¨®n, exactitud, especificidad, selectividad, precisi¨®n intermedia, dureza, robustez, estabilidad de la disoluci¨®n e idoneidad(AU)

Investigation of Antidiabetic, Antihyperlipidemic, and In Vivo Antioxidant Properties of Sphaeranthus indicus Linn. in Type 1 Diabetic Rats: An Identification of Possible Biomarkers.

The present investigation was aimed to study the antidiabetic, antihyperlipidemic, and in vivo antioxidant properties of the root of Sphaeranthus indicus Linn. in streptozotocin- (STZ-) induced type 1 diabetic rats. Administration of ethanolic extract of Sphaeranthus indicus root (EESIR) 100 and 200 mg/kg to the STZ-induced diabetic rats showed significant (P < .01) reduction in blood glucose and increase in body weight compared to diabetic control rats. Both the doses of EESIR-treated diabetic rats showed significant (P < .01) alteration in elevated lipid profile levels than diabetic control rats. The EESIR treatment in diabetic rats produced significant increase in superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and decrease in thiobarbituric acid reactive substances (TBARS) levels than diabetic control rats. Administration of EESIR 200 mg/kg produced significant (P < .01) higher antioxidant activity than EESIR 100 mg/kg. The high performance liquid chromatography (HPLC) analysis of EESIR revealed the presence of biomarkers gallic acid and quercetin. In conclusion, EESIR possess antidiabetic, antihyperlipidemic, and in vivo antioxidant activity in type 1 diabetic rats. Its antioxidant and lipid lowering effect will help to prevent diabetic complications, and these actions are possibly due to presence of above biomarkers.

Quality assessment of different marketed brands of Dasamoolaristam, an Ayurvedic formulation.

Arista is a classical Ayurvedic preparation that is typically used as a digestive and cardiotonic. The present Investigation evaluated five different brands of Dasamoolaristam available in the market as per WHO and Indian Pharmacopoeial specifications. Various physicochemical parameters such as alcohol-soluble extractive, water-soluble extractive, total ash, acid-insoluble ash, total solid, and alcohol content were determined. The present investigation reveals that all the preparations contain acceptable levels of alcohol (less than 12% v/v). However, the preparations were found to contain unacceptable limits of microbial load although all showed the absence of Escherichia coli, Salmonella species, and Staphylococcus aureus.

Evidence for a potential tumor suppressor role for the Na,K-ATPase beta1-subunit.

The Na,K-ATPase, consisting of two essential subunits (alpha, beta), plays a critical role in the regulation of ion homeostasis in mammalian cells. Recent studies indicate that reduced expression of the beta1 isoform (NaK-beta1) is commonly observed in carcinoma and is associated with events involved in cancer progression. In this study, we present evidence that repletion of NaK-beta1 in Moloney sarcoma virus-transformed Madin-Darby canine kidney cells (MSV-MDCK), a highly tumorigenic cell line, inhibits anchorage independent growth and suppresses tumor formation in immunocompromised mice. Additionally, using an in vitro cell-cell aggregation assay, we showed that cell aggregates of NaK-beta1 subunit expressing MSV-MDCK cells have reduced extracellular regulated kinase (ERK) 1/2 activity compared with parental MSV-MDCK cells. Finally, using immunohistochemistry and fully quantitative image analysis approaches, we showed that the levels of phosphorylated ERK 1/2 are inversely correlated to the NaK-beta1 levels in the tumors. These findings reveal for the first time that NaK-beta1 has a potential tumor-suppressor function in epithelial cells.

Unphosphorylated STAT6 contributes to constitutive cyclooxygenase-2 expression in human non-small cell lung cancer.

Cyclooxygenase-2 (COX-2) is frequently overexpressed in human cancers and contributes to the malignant phenotype. Our data indicate unphosphorylated signal transducers and activators of transcription 6 (STAT6) may transcriptionally upregulate COX-2 expression and protect against apoptosis in NSCLC cells. In A427 and H2122, NSCLC cell lines that constitutively express COX-2, only unphosphorylated STAT6 was detectable by western blot, thus, all of the following STAT6-dependent effects are attributed to the unphosphorylated protein. In both cell lines, small-interfering RNA-mediated knockdown of STAT6 or stable expression of dominant-negative STAT6 decreased COX-2 expression. In contrast, transfection with a phosphorylation-deficient mutant STAT6 increased COX-2 levels. Immunofluorescent staining revealed the presence of STAT6 in H2122 nuclei, suggesting a direct role in gene regulation for the unphosphorylated protein. Consistent with this hypothesis, unphosphorylated STAT6 increased luciferase expression from a COX-2 promoter reporter construct. STAT6 co-immunoprecipitated with the transcriptional co-activator, p300, and chromatin immunoprecipitation assays demonstrated that these proteins bind a consensus STAT6 binding site located within the COX-2 promoter. STAT6 DNA-binding specificity was confirmed by electrophoretic mobility shift assay. As COX-2 over-expression has been clearly linked to apoptosis resistance and other hallmarks of malignancy, these findings suggest a novel role of unphosphorylated STAT6 in the pathogenesis of non-small cell lung cancer.

Identification of protein kinase C as an intermediate in Na,K-ATPase beta-subunit mediated lamellipodia formation and suppression of cell motility in carcinoma cells.

We have shown that repletion of Na,K-ATPase Beta1-subunit (Na,K-Beta) in Moloney Sarcoma virus transformed MDCK (MSV-Na,K-Beta) cells induced lamellipodia and suppressed motility in a PI3-Kinase dependent manner. In this study, we provide evidence that decreased cell motility is due to increased attachment of Na,K-Beta expressing cells to the substratum. Treatment of MSV-Beta-GFP cells with bisindolylmalemide, a general Protein Kinase C (PKC) inhibitor, abolished PI3-Kinase activation and its down stream effects of Rac1 activation, binding of Na,K-Beta to annexin II, and suppression of cell motility and attachment. Thus, these studies unraveled that a PKC is involved upstream of PI3-Kinase in the suppression of Na,K-Beta mediated cell motility in carcinoma cells.