ANTIDIABETIC EFFECTS OF A SERIES OF CURCUMIN PYRAZOLES IN-VITRO

Objective: To investigate the antidiabetic potential of a series of curcumin pyrazole derivatives in-vitro.Methods: The in-vitro antidiabetic ability of curcumin pyrazole derivatives (3a-3e) was evaluated by the inhibition studies of the digestive enzymes;including alpha-amylase, rat intestinal alpha-glucosidase, and sucrase. The glucose lowering effect of the compounds was further assessed by theglucose uptake assay using a porcine diaphragm.Results: The curcumin pyrazole derivatives (3a-3e) showed a prominent inhibition of the enzymes studied. Compounds 3a and 3b were found tobe more pronounced in their hypoglycemic effects and 3b exhibited the maximum enzyme inhibitory activity. The glucose uptake through porcinediaphragm was extensively promoted by the compounds 3a and 3b than the other compounds in the series.Conclusion: The results suggest that the pyrazole derivatives of curcumin 3a and 3b have potent hypoglycemic properties. These compounds in theseries of curcumin pyrazole derivatives (3a-3e) can be selected for further in-vitro and in vivo studies.Keywords: Diaphragm, Insulin, Hypoglycemic, Enzyme inhibition

Molecular mechanisms involved in arsenic perception and tolerance in "Arabidopsis thaliana"

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 28-11-2014Arsenate is a toxic metalloid to living organisms. The incorporation of this toxic element to soil and water by anthropogenic and volcanic activities pose continuous threat to sessile organisms. Because of close chemical similarity between arsenate [As(V)] and phosphate (Pi), As(V) uses the Pi transporter to enter plant cell, which makes plants vulnerable for this metalloid. Suppression of Pi transporters is a strategy widely use by plants in nature to prevent arsenate uptake. However, closing Pi-transporters compromises phosphate availability and thus plant survival. Currently it is completely unknown, how plants manage this delicate situation preventing arsenic uptake without impairing phosphate acquisition. Here we found that in response to arsenate, the Pi/As(V) transporter PHT1;1 rapidly repressed its expression during initial 2 to 8 hrs of As(V) exposure. The early repression of the Pi-transporter is regulated by WRKY6, a repressor that is upregulated by As(V). The repression is coordinated by the degradation of PHR1, a transcriptional activator of PHT1;1. The repression is also accompanied by delocalization of the transporter from the plasma membrane. All these responses leads to an instant suppression of As(V) uptake. PHR1 degradation is mediated by ASK18, an SKP1-like protein that is transcriptionally regulated by As(V). Interestingly, ASK18 responsiveness depends on the arsenic detoxification status of plants. Furthermore, we show that plants reduce their endogenous cytokinin (CK) content as an early response to As(V) that leads to increase detoxification capacity. PHT1;1 repression and CKs reduction thus allows plants to coordinately reduce As(V) uptake and increased capacity to detoxify intracellular arsenic. Accordingly, transgenic lines with reduced levels of endogenous CKs shows remarkable arsenic tolerance phenotype. Surprisingly, after 24 hrs of As(V) exposure PHT1;1 repression is abolished and the expression is reactivated even in the presence of As(V). Here we found that PHT1;1 expression recovery is controlled by PHR1 stabilization that is also dependent on plant detoxification capacity. Overall the plants are able to adapt arsenic uptake to their detoxification capacity.El arsenato es un metaloide tóxico para los seres vivos. La incorporación de este elemento tóxico al suelo y al agua por actividades antropogénicas y volcánicas supuso una amenaza continua para los organismos sésiles. Debido a la estrecha similitud química del [As(V)] con el fosfato (Pi), el As(V) usa el transportador de Pi para entrar en las células vegetales, lo que hace a las plantas particularmente vulnerables a este metaloide. La supresión de los transportadores de Pi es una estrategia ampliamente utilizada por las plantas en la naturaleza para prevenir la absorción de arsenato. Sin embargo, el cierre de los transportadores de fosfato compromete la disponibilidad de este elemento y, por tanto, la supervivencia de la planta. Actualmente se desconoce el modo en que las plantas gestionan esta delicada situación, previniendo la absorción de arsenato sin dificultar la adquisición de fosfato. Hemos descubierto que el transportador de As(V)/Pi PHT1;1 reprime rápidamente su expresión en respuesta a arsenato, durante las 2 a 8 horas iniciales de exposición a dicho metaloide. La represión temprana del transportador de Pi es regulada por WRKY6, un represor cuya expresión aumenta en respuesta a As(V). Dicha represión está coordinada por la degradación de PHR1, un activador transcripcional de PHT1;1, y además está acompañada por la deslocalización del transportador de la membrana plasmática. Todas estas respuestas llevan a una supresión inmediata de la absorción de As(V). La degradación de PHR1 está mediada por ASK18, una proteína del tipo SKP1 cuya trascripción está regulada por As(V). Curiosamente, la respuesta de ASK18 depende del estado de detoxificación de arsénico de la planta. Además, hemos demostrado que las plantas reducen su contenido endógeno en citoquininas (CK) como respuesta temprana al As(V) lo cual produce un aumento en la capacidad de detoxificación. Por tanto la represión de PHT1;1 y la reducción de CKs permite a las plantas dar una respuesta coordinada, reduciendo la absorción de As(V) e incrementando la capacidad de detoxificación del arsénico intracelular. Así, líneas transgénicas con niveles reducidos de CKs endógenas muestran un fenotipo significativo de tolerancia a arsénico. Sorprendentemente, tras 24 horas de exposición a As(V), la represión de PHT1;1 se anula, reactivando su expresión incluso en presencia de As(V). Hemos encontrado que la recuperación de la expresión de PHT1;1 está controlada por la estabilización de PHR1, la cual también depende de la capacidad de detoxificación de la planta. En resumen, las plantas son capaces de adaptar la absorción de As(V) a su capacidad de detoxificació

ANTAGONISTIC EFFECTS OF COMBINATION OF FLAXSEED OIL AND SPIRULINA PLATENSIS OIL ON THEIR BIOLOGICAL PROPERTIES

Objective: To determine the composition of flaxseed oil and Spirulina platensis oil and to evaluate their biological activities either alone or in combination by in-vitro studies.Methods: Proximate analysis of flaxseeds and S. platensis powder, followed by compositional analysis of oil by gas chromatography was conducted. Cytotoxic studies were performed by MTT and trypan blue dye exclusion assay. Hypoglycemic studies were assessed by glucose uptake assay using porcine diaphragm and by gluconeogenesis assay using rat liver slices. Anti-denaturation assay and HRBC membrane assay were conducted to evaluate the anti-inflammatory properties.Results: α-linolenic acid (ALA) was the predominant component of flaxseed oil while γ-linolenic acid was found only in S. platensis oil. In all the experiments, S. platensis oil and flaxseed oil alone exhibited prominent anticancer, antidiabetic and anti-inflammatory activities. Whereas, all the above activities were reduced in the combination studies irrespective of the ratio of the mixture of oils used.Conclusion: The results clearly suggested that the combination of S. platensis oil and flaxseed oil does not have any synergistic properties, indeed the combination of oils induced antagonistic effects on their biological properties.Â

IN VITRO AND IN SILICO SCREENING OF 6-FLUORO-3-(PIPERIDIN-4-YL) BENZO[D]ISOXAZOLE DERIVATIVES FOR BLOOD BRAIN BARRIER PERMEABILITY

Blood Brian Barrier (BBB) is a physiologically and metabolically significant membrane that enables and limits the uptake of specific molecules by the brain, thus preserving the homeostasis within the Central nervous System (CNS).All the Antipsychotic drugs are permeable across the BBB, however most of First/Second generation antipsychotic drugs employed in the medical field are known to be associated with Extrapyramidal symptoms (EPS). Hence our interest is devoted in the development of novel antipsychotic drugs that ameliorate psychosis with least frequency of EPS. In our previous study the newly synthesized molecules, 6-fluoro-3-(piperidin-4-yl) benzo[d]isoxazole derivatives (S1-S4) which have shown positive alterations in chemical properties and biological activities demonstrating significant neuroleptic properties. However possibility and efficiency of these synthesized molecules to penetrate through the Blood Brain Barrier will be of utmost importance to determine its treatment efficacy. In this line of investigation, in the present study, in silico and in vitro methods are employed to evaluate the permeability in comparison with that of standard antipsychotic drugs, Resperidone and Haloperidol. In vitro analysis was done by Parallel artificial Membrane Permeability Assay (PAMPA) in PBS, where in permeability of S2(8.3x10-6) and S3(6.6x10-6) molecules showed higher permeability than known standards Warfarin(1x10-6), Resperidone(2.3x10-6) and Haloperidol(3.5x10-6), and the molecules exhibited recovery above 50% and in silico studies also showed high permeability rate with all the synthesized molecules. Thus we can conclude that, all the synthesized molecules have the potential to develop into promising Antipsychotic drugs.Key words: Antipsychotics, Blood Brain Barrier, EPS, PAMPA, In silicoÂ

IN VITRO ANTICANCER AND ANTI-LIPOXYGENASE ACTIVITIES OF CHIA SEED OIL AND ITS BLENDS WITH SELECTED VEGETABLE OILS

  Objective: To investigate the anticancer and anti-inflammatory potential of chia seed oil (CSO) and its blends in vitro.Methods: The in vitro cancer cytotoxic properties ability of CSO and its blends was evaluated by (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) and trypan blue assays using major cell lines including chronic myelogenous leukemia, HeLa and MCF-7 cells. The anti-inflammatory effect of the CSO and its blends was studied by assessing the anti-lipoxygenase (LOX) activity in vitro.Results: Incubation with CSO alone significantly inhibited the LOX activity. In the combination groups, CSO slightly increased the inhibition among blends of soybean and palmolein oils. In addition, CSO alone and in blends demonstrated potent and differential anticancer activity in MTT and trypan blue assays.Conclusion: Our results pave the way for the conclusion that CSO alone or in combination with the vegetable oils proves to be a healthy synergistic supplement or an adjuvant for the current diet. The supplementation with CSO is suggested for the modern lifestyle to delay or prevent the incidence of degenerative disorders

Targeting ie-1 gene by RNAi induces baculoviral resistance in lepidopteran cell lines and in transgenic silkworms

RNA interference (RNAi)-mediated viral inhibition has been used in a few organisms for eliciting viral resistance. In the present study, we report the use of RNAi in preventing baculovirus infection in a lepidopteran. We targeted the baculoviral immediate early-1 (ie-1) gene in both a transformed lepidopteran cell line and in the transgenic silkworm Bombyx mori L. Constitutive expression of double-stranded RNA was achieved by piggyBac-mediated transformation of Sf9 cell line with a transgene encoding double-stranded ie-1 RNA (dsie-1). Strong viral repression was seen at early stages of infection but subsequent recovery of viral proliferation was observed. In contrast, the same transgene inserted into the chromosomes of transgenic silkworms induced long-term inhibition of B. mori nucleopolyhedrovirus infection, with nearly 40% protection compared with nontransgenic animals. Protection was efficient at larval stages after oral infection with occlusion bodies or hemocoel injection of budded viruses. Virus injected pupae also displayed resistance. These results show that heritable RNAi can be used to protect silkworm strains from baculovirus infection

Differential attainment and recruitment to Intensive Care Medicine training in the UK, 2018–2020

Background Differences exist among doctors in examination performance, clinical and academic career progression, and prevalence of performance assessment by professional regulatory bodies. Some of these differences have been reported in relation to individual characteristics. The purpose of this study is to establish whether any specific individual characteristics are associated with performance in selection for entry into specialty training in Intensive Care in the United Kingdom. Methods We evaluated data of 509 candidates from the national recruitment rounds of 2018/19 and 2019/20. The outcome evaluated was “success at interview". Variables reaching statistical significance at univariate logistic regression analysis were fed in the multivariable analysis to identify independent predictors of success, with additional exploratory analyses performed, where indicated. Results The candidates’ median age was 31.5 (interquartile range, IQR 30–33.7) years, 324 (63.7%) were male, 256 (50.3%) not married/in civil partnership, 6 (1.2%) pregnant. The majority (316, 62.1%) were White British, 99 (19.5%) of Asian background, other ethnicities represented less than 20% of the sample. Of the 509 candidates, 155 (30.5%) were Atheist, 140 (27.5%) Christian; most were heterosexual (440, 86.4%); 432 (84.9%) reported no disability, while 4 (0.8%) had a minor and 1 (0.2%) had a major disability; 432 (84.9%) candidates held a UK medical degree; 77 (15.1%) a non-UK degree. At univariate logistic regression analysis (LRA) multiple factors were found to be associated with a lower likelihood of success, the strongest being an international medical graduate (IMG, holding a non-UK medical degree); others were increasing age, male gender, being married, Asian or mixed ethnicity, specific religious beliefs (Buddhism, Islam and Hinduism). After feeding all factors significant at univariate analysis, the only two retained as independent predictors at multivariable regression were Asian ethnicity and holding a non-UK degree. Asian UK graduates success rate was 92.7%, comparable to the national average of 92.3%, the Asian IMGs success rate was significantly lower, at 45.5%. Conclusions As the imbalances seen within the candidates of Asian background are explained by considering the country of primary medical training, the variations in performance is likely to reflect differences in training systems and understanding of the UK NHS

Management of Critically Ill Patients of Hoiiow Viscus Perforation by Mini Laprotomy Followed by Regular Laprotomy

Background: Gastrointestinal perforation is a common abdominal emergency having high morbidity and mortality. Surgery plays an important role in the management of perforation.Methods: 20 cases of critically ill patients with abdominal compartment syndrome are taken for the study. Abdominal compartment pressure more than 25 cm of NS are taken for the study. Study period is from January 2016 to February 2020.Patients are managed differently .mini laparotomy done under local anesthesia, abdomen decompressed. later regular surgery done. Results: Results obtained in this study are analyzed. In Majority of patients BP improved by10to20mmhg after mini laparotomy done under local anesthesia and urine out put also star ted increasing. Duodenal ulcer perforation is the commonest cause and then typhoid ulcer perforation. Smoking and alcohol intake are the common etiology for D.U. perforation. In this study mortality is 5%.Conclusion :Though mortality is high in critically ill patients of hollow viscus perforation, here probably mortality has reduced due to mini laprotomy done under local anesthesia. More studies are required to know more about This study

Interproximal bone loss assessment: Comparison of conventional and digital radiographs.

Intraoral radiographs are important diagnostic tools that play a major role in diagnosis, treatment plan and prognosis of periodontal diseases. This study assesses the accuracy of intraoral periapical (IOPA) and radiovisiography (RVG) radiographs in detection of interproximal alveolar bone loss using intrasurgical (IS) measurements as the gold standard. Interproximal alveolar bone loss were measured in 23 patients with moderate to severe periodontitis, using digital vernier caliper for IOPA and linear measurement tool inbuilt in RVG system, from radiographs taken with standardized techniques which were compared with the surgical readings. 106 interproximal sites were measured in IOPA, RVG radiographs and compared with IS method. Paired t test showed significant difference in bone levels between the systems. A contingency analysis of categorical bone levels (early, moderate & advanced) showed significant differences in the imaging systems. Overall radiographic assessment of interproximal bone loss by either IOPA or RVG radiographs shows no total agreement in comparison with IS measurements, although it was found that both conventional and digital radiographs are of use in interproximal bone loss assessment but in different similarities