On the macro-micro-morphology of organs of host invasion in hemiparasite Helicanthes elasticus (Desv.) Danser

Loranthaceae family includes hemiparasitic members which are seen invading a wide range of commercial crops. Helicanthes elasticus (Desv.) Danser is very common on mango trees. Though parasitic in nature, this mistletoe is also medicinally important as fetoprotective, against vesicular calculi and kidney infections. This study is an attempt to document macro-microscopical features of parasitic root, fruit and host-mistletoe tissue interaction in the haustorium of H. elasticus growing on mango stems. Collection, preservation, sectioning, staining and photomicrography of the root, fruit and host-mistletoe union were done as per standard methodologies of anatomical studies. Though there is resemblance to the normal roots in morphology as well as anatomy, the microscopic finding of large number of branched stone cells in the roots is interesting. The morpho-anatomical features recorded would help in understanding the infection biology of this mistletoe. The eradication during the earlier stages of its establishment from seed or from the root creeping over the surface of the host can help in controlling this parasite infection on commercially important host plants.     

The effect of high-fat diet and inhibition of ceramide production on insulin action in liver

Liver, as one of the most important organs involved in lipids and glucose metabolism, is perceived as a key tissue for pharmacotherapy of insulin resistance (IRes) and type 2 diabetes. Ceramides (Cer) are biologically active lipids, which accumulation is associated with the induction of muscle IRes. We sought to determine the role of intrahepatic bioactive lipids production on insulin action in liver of insulin-resistant rats and after myriocin administration. The experiments were conducted on male Wistar rats divided into three groups: Control, fed high-fat diet (HFD), and fed HFD and treated with myriocin (HFD/Myr). Before sacrifice, the animals were infused with a [U-¹³C]palmitate to calculate lipid synthesis rate by means of tracer incorporation technique in particular lipid groups. Liver Cer, diacylglycerols (DAG), acyl-carnitine concentration, and isotopic enrichment were analyzed by LC/MS/MS. Proteins involved in lipid metabolism and insulin pathway were analyzed by western blot analysis. An OGTT and ITT was also performed. HFD-induced IRes and increased both the synthesis rate and the content of DAG and Cer, which was accompanied by inhibition of an insulin pathway. Interestingly, myriocin treatment reduced synthesis rate not only of Cer but also DAG and improved insulin sensitivity. We conclude that the insulin-sensitizing action of myriocin in the liver is a result of the lack of inhibitory effect of lipids on the insulin pathway, due to the reduction of their synthesis rate. This is the first study showing how the synthesis rate of individual lipid groups in liver changes after myriocin administration.     

Identification of BARD1 splice-isoforms involved in human trophoblast invasion

The tumor suppressor protein BARD1, originally discovered as BRCA1-binding protein, acts in conjunction with BRCA1 as ubiquitin ligase. BARD1 and BRCA1 form a stable heterodimer and dimerization, which is required for most tumor suppressor functions attributed to BRCA1. In addition, BARD1 has BRCA1-independent functions in apoptosis, and a role in control of tissue homeostasis was suggested. However, cancer-associated mutations of BARD1 are rare; on the contrary, overexpression of truncated BARD1 was found in breast and ovarian cancer and correlated with poor prognosis. Here we report that human cytotrophoblasts, which show a strong similarity with cancer cells in respect of their invasive behavior and capacity of matrix metalloprotease production, overexpress isoforms of BARD1 derived from differential splicing. We demonstrate that expression of BARD1 and its isoforms is temporally and spatially regulated by human chorionic gonadotropin and by hypoxia, both factors known to regulate the invasive phase and proliferation of cytotrophoblasts. Interestingly, we found a subset of BARD1 isoforms secreted by cytotrophoblasts. BARD1 repression by siRNAs, mitigates the interference of cytotrophoblasts with cell adhesion of collagen matrix-dependent epithelial cells, suggesting a role of BARD1 isoforms in extracellular matrix remodelling and in cytotrophoblasts invasion.     

Divergent responses of different endothelial cell types to infection with Candida albicans and Staphylococcus aureus

Endothelial cells are important in the pathogenesis of bloodstream infections caused by Candida albicans and Staphylococcus aureus. Numerous investigations have used human umbilical vein endothelial cells (HUVECs) to study microbial-endothelial cell interactions in vitro. However, the use of HUVECs requires a constant supply of umbilical cords, and there are significant donor-to-donor variations in these endothelial cells. The use of an immortalized endothelial cell line would obviate such difficulties. One candidate in this regard is HMEC-1, an immortalized human dermal microvascular endothelial cell line. To determine if HMEC-1 cells are suitable for studying the interactions of C. albicans and S. aureus with endothelial cells in vitro, we compared the interactions of these organisms with HMEC-1 cells and HUVECs. We found that wild-type C. albicans had significantly reduced adherence to and invasion of HMEC-1 cells as compared to HUVECs. Although wild-type S. aureus adhered to and invaded HMEC-1 cells similarly to HUVECs, an agr mutant strain had significantly reduced invasion of HMEC-1 cells, but not HUVECs. Furthermore, HMEC-1 cells were less susceptible to damage induced by C. albicans, but more susceptible to damage caused by S. aureus. In addition, HMEC-1 cells secreted very little IL-8 in response to infection with either organism, whereas infection of HUVECs induced substantial IL-8 secretion. This weak IL-8 response was likely due to the anatomic site from which HMEC-1 cells were obtained because infection of primary human dermal microvascular endothelial cells with C. albicans and S. aureus also induced little increase in IL-8 production above basal levels. Thus, C. albicans and S. aureus interact with HMEC-1 cells in a substantially different manner than with HUVECs, and data obtained with one type of endothelial cell cannot necessarily be extrapolated to other types.     

New development for combined bioscouring and bleaching of cotton-based fabrics

A thorough investigation into conditions appropriate for effecting combined eco-friendly bioscouring and/or bleaching of cotton-based fabrics was undertaken. Fabrics used include cotton, grey mercerized cotton, cotton/polyester blend 50/50 and cotton/polyester blend 35/65. The four cotton-based fabric were subjected to bioscouring by single use of alkaline pectinase enzymes or by using binary mixtures of alkaline pectinase and cellulase enzymes under a variety of conditions. Results of bioscouring show that, the bioscoured substrates exhibit fabrics performances which are comparable with these of the conventional alkali scouring. It has been also found that, incorporation of ethylenediaminetetraacetic acid (EDTA) in the bioscouring with mixture from alkaline pectinase and cellulase improves the performance of the bioscoured fabrics. Addition of β-cyclodextrin to the bioscouring solution using alkaline pectinase in admixtures with cellulase acts in favor of technical properties and performance of the bioscoured fabrics. Concurrent bioscouring and bleaching by in situ formed peracetic acid using tetraacetylethylenediamine (TAED) and H₂O₂ was also investigated. The results reveal unequivocally that the environmentally sound technology brought about by current development is by far the best. The new development involves a single-stage process for full purification/preparation of cotton textiles. The new development at its optimal comprises treatment of the fabric with an aqueous formulation consisting of alkaline pectinase enzyme (2 g/L), TAED (15 g/L), H₂O₂ (5 g/L), nonionic wetting agent (0.5 g/L) and sodium silicate (2 g/L). The treatment is carried out at 60 °C for 60 min. Beside the advantages of the new development with respect to major technical fabric properties, it is eco-friendly and reproducible. This advocates the new development for mill trials.     

Isolation of antifungal compounds from someZygophyllum species and their bioassay against two soil-borne plant pathogens

Roots and aerial shoots ofZygophyllum coccineum L.,Z. album L. andZ. dumosum Boiss were extracted with solvents of increasing polarity and tested against spore germination ofVerticellium albo-atrum andFusarium oxysporum f.sp.lycopersici. The effectiveness of each extract depends on the solvent, plant species, organ and test fungus. Petroleum ether extracts were ineffective as fungistatics while the methanol extracts possessed a high inhibitory effect toward spore germination suppression, depending on the fungus and plant species. Water or ethyl acetate root extracts ofZ. album caused a marked suppression of spore germination of the test fungi. Six saponins were isolated from the effective extracts on silica gel and Sephadex columns. They were identified by acid hydrolysis, alkaline saponification, IR and FAB MS. Quinovic acid was the dominant aglycone. Kaempferol was isolated from the flavonoid fraction ofZ. dumosum for the first time. Some of the isolated saponins and kaempferol provided successful control against the test plant pathogens under greenhouse experiment when used as seed treatment.     

The influence of laparoscopic adjustable gastric banding and laparoscopic sleeve gastrectomy on weight loss, plasma ghrelin, insulin, glucose and lipids

The aim of this study was to assess the impact of laparoscopic gastric banding and laparoscopic sleeve gastrectomy on the concentration of ghrelin, insulin, glucose, triglycerides, total and HDL-cholesterol, as well as AST and ALT levels in plasma in patients with obesity. The research includes 200 patients operated using LAGB (34 men average age 37.0 ± 12.6 years and 66 women average age 39.18 ± 12.17 years) and LSG (48 men average age 47.93 ± 9.24 years and 52 women, 19 ± 9.33 years). The percentage of effective weight loss, effective BMI loss, concentration of ghrelin, insulin, glucose, triglycerides, total cholesterol, HDL-cholesterol, LDL-cholesterol, ALT, AST and HOMA IR values was taken preoperatively and at 7th day, 1 month, 3 and 6 months after surgery. Both after LSG and after LAGB, statistically significant reduction in BMI, serum insulin, glucose and HOMA IR was noticed in comparison to the preoperative values. Post LAGB, patients showed an increase of ghrelin, while LSG proved ghrelin decreased. Correlations between glucose and BMI loss, and between insulin and BMI loss in both cases are more favorable in the LSG group. Lipid parameters, AST and ALT have undergone declines or increases in the particular time points. Both techniques cause weight loss and this way lead to changes in the concentration of ghrelin, as well as to the improvement of insulin, glucose, cholesterol and triglycerides metabolism. They reduce metabolic syndrome and multiple comorbidities of obesity.     

Humanized beta-thalassemia mouse model containing the common IVSI-110 splicing mutation

Splicing mutations are common causes of beta-thalassemia. Some splicing mutations permit normal splicing as well as aberrant splicing, which can give a reduced level of normal beta-globin synthesis causing mild disease (thalassemia intermedia). For other mutations, normal splicing is reduced to low levels, and patients are transfusion-dependent when homozygous for the disease. The development of therapies for beta-thalassemia will require suitable mouse models for preclinical studies. In this study, we report the generation of a humanized mouse model carrying the common IVSI-110 splicing mutation on a BAC including the human beta-globin ((hu)beta-globin) locus. We examined heterozygous murine beta-globin knock-out mice ((mu)beta(th-3/+)) carrying either the IVSI-110 or the normal (hu)beta-globin locus. Our results show a 90% decrease in (hu)beta-globin chain synthesis in the IVSI-110 mouse model compared with the mouse model carrying the normal (hu)beta-globin locus. This notable difference is attributed to aberrant splicing. The humanized IVSI-110 mouse model accurately recapitulates the splicing defect found in comparable beta-thalassemia patients. This mouse model is available as a platform for testing strategies for the restoration of normal splicing.