SENNA PLANT INDUCES DISRUPTION ON THE MITOCHONDRIA OF HYMENOLEPIS DIMINUTA

Objective: The present study aims at observing the effects of three species of Senna plants, viz. Senna alata, S. alexandrina and S. occidentalis on the ultrastructure of the mitochondria of the tapeworm, Hymenolepis diminuta.Methods: Worms were treated with leaf extracts of the three plant species with a standard dose concentration of 40 mg/ml and keeping one group of parasites in phosphate buffer saline (PBS) as a control. The parasites from control and treated medium were simultaneously removed after the loss of motility and fixed in 3% gluteraldehyde. They were processed for ultramicrograph observations of the worm's mitochondria with special reference to shape and cytoplasm through transmission electron microscopy (TEM).Results: The study showed loss of architecture in the outer mitochondrial membrane. The inner membrane became distorted with inconspicuous cristae and matrix became lucent in all plant treated worms compared to control. Amongst the three plants, S. alexandrina showed overall distortion in the shape leading to bloating of mitochondria.Conclusion: The observations depict pronounced alterations in the structure of mitochondria, thus signifying depletion of energy synthesis in the parasite. Senna plant could, therefore, be a potent anthelmintic alternative

IN VITRO ANTHELMINTIC EFFECTS OF SENNA OCCIDENTALIS (L.) LINK (LEGUMINOSAE) ON RAT TAPEWORM HYMENOLEPIS DIMINUTA

Objective: This study was conducted to evaluate the anthelmintic activity of ethanolic leaf extract of Senna occidentalis on rat tapeworm Hymenolepis diminuta.Methods: Adult H. diminuta worms were obtained from laboratory maintained infection in rats. Different concentrations of crude ethanolic leaf extracts of S. occidentalis were tested in vitro on H. diminuta and praziquantel (PZQ) was used as reference drug. The efficacy of plant was assessed on the basis of motility of the worms. The anthelmintic effects were also evaluated using scanning and transmission electron microscopy.Results: A dose dependent efficacy was observed in all the experiments. At the highest concentration (80 mg/ml), the mortality of worms was recorded in 12.82±0.24 hrs. The scanning electron microscope observations of extract treated worms showed irrevocable destruction all over the body tegument,which was accompanied with sloughing of microthriches and shrinkage of scolex. Whereas, the transmission electron micrography of treated worms revealed destruction of the syncytial layer along with sparse cytoplasmic cytons and depletion of parenchymatous layer. Exposure of basal lamina, decrease in an electron lucency in the nucleus and intense vacuolization was also observed in our study.Conclusion: In view of these observations, S. occidentalis may be regarded as potential anthelmintic plant and it could be exploited further for the development of a novel anthelmintic.Â

Affinity binding of non-histone chromatin proteins to the X chromosome of Drosophila by in situ chromatin reconstitution and its significance

Cytophotometric analysis of the in situ binding affinity of non-histone chromosomal protein (NHCP) to the polytenic X chromosome and autosome of Drosophila melanogaster has been carried out using Feulgen-Napthol Yellow S staining technique. The results reveal that the mean transformed absorbance ratio (male:female) with a 547nm interference band filter for the two specific segments of the X chromosome is close to 0-5, while for a specific segment of an autosome it is close to l"0, in the two sets of control; namely, the positive control (no treatment) and the negative control (treated with 1 M-urea+2M-NaCl) as well as in the reconstituted chromosomal preparations, which received 1 M-urea+2M-NaCl and the NHCP isolated from D. melanogaster. In contrast, the transformed absorbance ratios (male:female) with a 433 nm interference band filter yielded an interestingly different result. The ratios with a 433 nm filter for the X chromosome segments are significantly greater than 0-5 in all three sets of experiments. This finding by itself suggests that the NHCP binding affinity is dissimilar for the X chromosomes of male and female. When the 433 to 547 nm absorbance ratios were compared among the three sets, the data clearly revealed that in both positive control and NHCP reconstituted samples, the absorbance ratios (i.e. 433:547 nm) are significantly different between X chromosomes from males and those from females, while they are different between autosomes from males and females. The ratios are also not significantly different between male and female, either for the X chromosome or for the autosome in the negative control. These findings, therefore, suggest that there is a stronger binding affinity of NHCP for the male X chromosome of Drosophila, and reinstate the view that the X chromosomal hyperactivity in male Drosophila is the consequence of a regulated organizational change in the DNA template

Ultrahigh temperature and strain hybrid integrated sensor system based on Raman and femtosecond FBG inscription in a multimode gold-coated fiber

In this paper, a novel approach for hybrid systems combining Raman distributed sensors with fiber Bragg grating (FBG) sensors to carry out distributed and quasi-distributed temperature/strain measurements was proposed and experimentally demonstrated. Three FBGs were inscribed by the point-by-point technique in a simple setup for type I femtosecond inscription in a pure silica core multimode gold-coated fiber. Employing a single fiber, temperatures up to 600 °C and strains up to 4144 µE approximately were measured simultaneously and without interferences between both distributed and point measurements. Moreover, a new calibration technique was implemented to calibrate the distributed temperature system using the FBG measurements as reference.Spanish Government FEDER Funds (TEC2016-76021-C2-2-R)

Curative Effect of 18β-Glycyrrhetinic Acid in Experimental Visceral Leishmaniasis Depends on Phosphatase-Dependent Modulation of Cellular MAP Kinases

We earlier showed that 18β-glycyrrhetinic acid (GRA), a pentacyclic triterpenoid from licorice root, could completely cure visceral leishmaniasis in BALB/c mouse model. This was associated with induction of nitric oxide and proinflammatory cytokine production through the up regulation of NF-κB. In the present study we tried to decipher the underlying cellular mechanisms of the curative effect of GRA. Analysis of MAP kinase pathways revealed that GRA caused strong activation of p38 and to a lesser extent, ERK in bone marrow-derived macrophages (BMDM). Almost complete abrogation of GRA-induced cytokine production in presence of specific inhibitors of p38 and ERK1/2 confirmed the involvement of these MAP kinases in GRA-mediated responses. GRA induced mitogen- and stress-activated protein kinase (MSK1) activity in a time-dependent manner suggested that GRA-mediated NF-κB transactivation is mediated by p38, ERK and MSK1 pathway. As kinase/phosphatase balance plays an important role in modulating infection, the effect of GRA on MAPK directed phosphatases (MKP) was studied. GRA markedly reduced the expression and activities of three phosphatases, MKP1, MKP3 and protein phosphatase 2A (PP2A) along with a substantial reduction of p38 and ERK dephosphorylation in infected BMDM. Similarly in the in vivo situation, GRA treatment of L. donovani-infected BALB/c mice caused marked reduction of spleen parasite burden associated with concomitant decrease of individual phosphatase levels. However, activation of kinases also played an important role as the protective effect of GRA was significantly abrogated by pharmacological inhibition of p38 and ERK pathway. Curative effect of GRA may, therefore, be associated with restoration of proper cellular kinase/phosphatase balance, rather than modulation of either kinases or phosphatases

Rectal gel application of Withania somnifera root extract expounds anti-inflammatory and muco-restorative activity in TNBS-induced Inflammatory Bowel Disease

<p>Abstract</p> <p>Background</p> <p>Inflammatory Bowel Disease (IBD) is marked with chronic inflammation of intestinal epithelium driven by oxidative stress. Traditional treatments with plant extracts gained renewed interest due to their ability to ameliorate the multi factorial conditions like inflammation. We investigated the beneficial effects of <it>Withania somnifera </it>in Trinitro Benzyl Sulfonic Acid (TNBS) induced experimental IBD through a rectally applicable formulation.</p> <p>Methods</p> <p>The study included (i) preparation of gel formulation from aqueous <it>Withania somnifera </it>root extract (WSRE), (ii) biochemical assays to determine its performance potential, (iii) testing of formulation efficacy in TNBS-induced IBD rat model, and (iv) histo-patholgical studies to assess its healing and muco-regenerative effect in IBD-induced rats. For this purpose, concentration dependant antioxidant activity of the extracts were evaluated using biochemical assays like (a) inhibition of lipid peroxidation, (b) NO scavenging, (c) H₂O₂scavenging, and (d) ferric reducing power assay.</p> <p>Results</p> <p>The extract, at 500 μg/ml, the highest concentration tested, showed 95.6% inhibition of lipid peroxidation, 14.8% NO scavenging, 81.79% H₂O₂scavenging and a reducing capacity of 0.80. The results were comparable with standard antioxidants, ascorbic acid and curcumin. WSRE treatment positively scored on histopathological parameters like necrosis, edema, neutrophil infiltration. The post treatment intestinal features showed restoration at par with the healthy intestine. In view of these results, gel formulation containing an aqueous extract of <it>W. somnifera</it>, prepared for rectal application was tested for its anti-inflammatory activity in TNBS-induced rat models for IBD. Commercially available anti-inflammatory drug Mesalamine was used as the standard in this assay.</p> <p>Conclusions</p> <p>Dose of the rectal gel applied at 1000 mg of WSRE per kg rat weight showed significant muco-restorative efficacy in the IBD-induced rats, validated by histo-pathological studies.</p

Fission Yeast Cells Undergo Nuclear Division in the Absence of Spindle Microtubules

Through a previously undescribed mechanism, fission yeast cells can undergo nuclear division and enter the next cell cycle, even in the absence of spindle microtubules

Comparing algorithms for automated vessel segmentation in computed tomography scans of the lung: the VESSEL12 study

The VESSEL12 (VESsel SEgmentation in the Lung) challenge objectively compares the performance of different algorithms to identify vessels in thoracic computed tomography (CT) scans. Vessel segmentation is fundamental in computer aided processing of data generated by 3D imaging modalities. As manual vessel segmentation is prohibitively time consuming, any real world application requires some form of automation. Several approaches exist for automated vessel segmentation, but judging their relative merits is difficult due to a lack of standardized evaluation. We present an annotated reference dataset containing 20 CT scans and propose nine categories to perform a comprehensive evaluation of vessel segmentation algorithms from both academia and industry. Twenty algorithms participated in the VESSEL12 challenge, held at International Symposium on Biomedical Imaging (ISBI) 2012. All results have been published at the VESSEL12 website http://vessel12.grand-challenge.org. The challenge remains ongoing and open to new participants. Our three contributions are: (1) an annotated reference dataset available online for evaluation of new algorithms; (2) a quantitative scoring system for objective comparison of algorithms; and (3) performance analysis of the strengths and weaknesses of the various vessel segmentation methods in the presence of various lung diseases.Rudyanto, RD.; Kerkstra, S.; Van Rikxoort, EM.; Fetita, C.; Brillet, P.; Lefevre, C.; Xue, W.... (2014). Comparing algorithms for automated vessel segmentation in computed tomography scans of the lung: the VESSEL12 study. Medical Image Analysis. 18(7):1217-1232. doi:10.1016/j.media.2014.07.003S1217123218

Functional Analysis of the Kinome of the Wheat Scab Fungus Fusarium graminearum

As in other eukaryotes, protein kinases play major regulatory roles in filamentous fungi. Although the genomes of many plant pathogenic fungi have been sequenced, systematic characterization of their kinomes has not been reported. The wheat scab fungus Fusarium graminearum has 116 protein kinases (PK) genes. Although twenty of them appeared to be essential, we generated deletion mutants for the other 96 PK genes, including 12 orthologs of essential genes in yeast. All of the PK mutants were assayed for changes in 17 phenotypes, including growth, conidiation, pathogenesis, stress responses, and sexual reproduction. Overall, deletion of 64 PK genes resulted in at least one of the phenotypes examined, including three mutants blocked in conidiation and five mutants with increased tolerance to hyperosmotic stress. In total, 42 PK mutants were significantly reduced in virulence or non-pathogenic, including mutants deleted of key components of the cAMP signaling and three MAPK pathways. A number of these PK genes, including Fg03146 and Fg04770 that are unique to filamentous fungi, are dispensable for hyphal growth and likely encode novel fungal virulence factors. Ascospores play a critical role in the initiation of wheat scab. Twenty-six PK mutants were blocked in perithecia formation or aborted in ascosporogenesis. Additional 19 mutants were defective in ascospore release or morphology. Interestingly, F. graminearum contains two aurora kinase genes with distinct functions, which has not been reported in fungi. In addition, we used the interlog approach to predict the PK-PK and PK-protein interaction networks of F. graminearum. Several predicted interactions were verified with yeast two-hybrid or co-immunoprecipitation assays. To our knowledge, this is the first functional characterization of the kinome in plant pathogenic fungi. Protein kinase genes important for various aspects of growth, developmental, and infection processes in F. graminearum were identified in this study