Overexpression of fatty acid synthase in Middle Eastern epithelial ovarian carcinoma activates AKT and Its inhibition potentiates cisplatin-induced apoptosis

Fatty acid synthase (FASN), the enzyme responsible for de novo synthesis of fatty acids, has been shown to be deregulated in several cancers, including epithelial ovarian carcinoma (EOC). In this study, we investigated the function of the FASN signaling pathway in a large series of Middle Eastern EOC patient samples, a panel of cell lines and nude mouse model. Using immunohistochemistry, we detected overexpression of FASN in 75.5% (114/151) of the tumor samples. Overexpression of FASN was associated significantly with tumor proliferative marker Ki-67 (P = 0.0009), activated AKT (P = 0.0117) and XIAP (P = 0.0046). Treatment of EOC cell lines with C-75, a selective inhibitor of FASN, caused inhibition of EOC cell viability via induction of apoptosis. Inhibition of FASN by C-75 led apoptosis via the mitochondrial pathway. FASN inhibition caused downregulation of activated AKT and its downstream targets. In addition, inhibition by FASN siRNA caused downregulation of FASN and activation of caspases, suggesting the role of FASN in C-75 mediated apoptosis. Furthermore, treatment of EOC cells with subtoxic doses of C-75 augmented the effect of cisplatin-mediated induction of apoptosis. Finally, treatment of EOC cell line xenografts with a combination of C-75 and cisplatin resulted in growth inhibition of tumors in nude mice through downregulation of FASN and activation of caspases. Altogether, our results show overexpression of FASN in Middle Eastern EOC, suggesting that FASN may be a potential therapeutic target in a subset of EOC, alone or in combination with other conventional chemotherapeutic agents.     

Primary human epithelial cell culture system for studying interactions between female upper genital tract and sexually transmitted viruses, HSV-2 and HIV-1

Evidence from clinical and epidemiological studies indicates that women are disproportionately susceptible to sexually transmitted viral infections. To understand the underlying biological basis for this increased susceptibility, more studies are needed to examine the acute events in the female reproductive tract following exposure to viruses during sexual transmission. The epithelial lining of the female reproductive tract is the primary barrier that sexually transmitted viruses, such as HIV-1 and HSV-2 need to infect or traverse, in order to initiate and establish productive infection. We have established an ex-vivo primary culture system to grow genital epithelial cells from upper reproductive tract tissues of women. Using these cultures, we have extensively examined the interactions between epithelial cells of the female genital tract and HSV-2 and HIV-1. In this review, we describe in detail the experimental protocol to grow these cultures, monitor their differentiation and inoculate with HSV-2 and HIV-1. Prospective use of these cultures to re-create the microenvironment in the reproductive tract is discussed.     

Role of transgenic plants in agriculture and biopharming

At present, environmental degradation and the consistently growing population are two main problems on the planet earth. Fulfilling the needs of this growing population is quite difficult from the limited arable land available on the globe. Although there are legal, social and political barriers to the utilization of biotechnology, advances in this field have substantially improved agriculture and human life to a great extent. One of the vital tools of biotechnology is genetic engineering (GE) which is used to modify plants, animals and microorganisms according to desired needs. In fact, genetic engineering facilitates the transfer of desired characteristics into other plants which is not possible through conventional plant breeding. A variety of crops have been engineered for enhanced resistance to a multitude of stresses such as herbicides, insecticides, viruses and a combination of biotic and abiotic stresses in different crops including rice, mustard, maize, potato, tomato, etc. Apart from the use of GE in agriculture, it is being extensively employed to modify the plants for enhanced production of vaccines, hormones, etc. Vaccines against certain diseases are certainly available in the market, but most of them are very costly. Developing countries cannot afford the disease control through such cost-intensive vaccines. Alternatively, efforts are being made to produce edible vaccines which are cheap and have many advantages over the commercialized vaccines. Transgenic plants generated for this purpose are capable of expressing recombinant proteins including viral and bacterial antigens and antibodies. Common food plants like banana, tomato, rice, carrot, etc. have been used to produce vaccines against certain diseases like hepatitis B, cholera, HIV, etc. Thus, the up- and down-regulation of desired genes which are used for the modification of plants have a marked role in the improvement of genetic crops. In this review, we have comprehensively discussed the role of genetic engineering in generating transgenic lines/cultivars of different crops with improved nutrient quality, biofuel production, enhanced production of vaccines and antibodies, increased resistance against insects, herbicides, diseases and abiotic stresses as well as the safety measures for their commercialization.     

The Path from Skin to Brain: Generation of Functional Neurons from Fibroblasts

Cell fate reprogramming makes possible the generation of new cell types from healthy adult cells to replace those lost or damaged in disease. Additionally, reprogramming patient cells into specific cell types allows for drug screening and the development of new therapeutic tools. Generation of new neurons is of particular interest because of the potential to treat diseases of the nervous system, such as neurodegenerative disorders and spinal cord injuries, with cell replacement therapy. Recent advances in cell fate reprogramming have led to the development of novel methods for the direct conversion of fibroblasts into neurons and neural stem cells. This review will highlight the advantages of these new methods over neuronal induction from embryonic stem cells and induced pluripotent stem cells, as well as outline the limitations and the potential for future applications.     

Morphological indicators of salinity stress and their relation with osmolyte associated redox regulation in mango cultivars

Soil salinization is a global issue impeding horticulture production and is approaching an alarming status due to climate change and urbanization. Breeding salt-tolerant rootstock varieties is an ideal strategy to mitigate stress due to salinity in mango and other perennial fruit species. Stress combating strategies employed by seedlings of 7 mango were studied under saline conditions (200 mM NaCl, EC: > 4.0 dSm⁻¹, pH 8.5) in pot experiments. Significantly high accumulation of proline (19.07 µg g⁻¹ FW in Bappakai), glycine betaine (55.11 µg g⁻¹ FW in 13–1), and total sugars (17.33 mg g⁻¹ FW in Kurukkan) were found to be the common mechanism employed by the tolerant cultivars to counter the osmotic stress, under suboptimal conditions. Non-enzymatic antioxidants viz., tannins (17.18 mg g⁻¹), phenols (18.68 mg g⁻¹), and anthocyanins (1.59 mg g⁻¹) were increased in seedling of "13–1", the salt-resistant cultivar from Israel. Reactive oxygen species (ROS) regulation by increased activity of superoxide dismutase and catalase in the two polyembryonic cultivars of Indian origin (Kurukkan and Nekkare) suggests their potential use as rootstocks to combat oxidative stress. The tolerance index of various cultivars was calculated by averaging the scores of morphological stress indicators, and its correlation with studied parameters suggests that salinity resilience is more tightly linked to enhanced catalase accumulation (r² = 0.8361) that is reduced ionic stress. This evidence assign the role of osmotic stress alleviation and redox regulation in salt tolerance mechanism operational in native Indian cultivars, Nekkare and Kurukkan at par with known salt tolerant rootstocks.

     

Air-borne fungi at Doha, Qatar

Thirty-five genera and 73 species, were identifiedfrom 312 daily exposures set up during theperiod March 1997–March 1998. The total fungalcatch exhibited two peaks in July and December1997 and a trough in February 1998. Cladosporium (6 spp. 40.1% of total fungi),Alternaria (4 spp., 21%) andUlocladium (4 spp., 9.2%) were the maincomponents of air-borne fungi, and thecommonest species were Cladosporium.sphaerospermum (29.7%), C.cladosporioides (6.9%), Alternaria.alternata (13.9%) and U. atrum (5%).The predominance of these dark-coloured fungiin air is discussed and is attributed to one orboth of two hypotheses. Aspergillus (9spp., 4.3%) and Penicillium (8 spp.,3.95%) came next and were represented mainlyby A.niger (1.3%) andP. chrysogenum (2.4%).Spore showers of C.cladosporioides, C. sphaerospermum, Penicillium chrysogenum and Myrotheciumverrucaria were noticed with no regularseasonal pattern.The monthly number of species ranalmost parallel to the total count of fungi.The broadest species spectrum (25–29 spp.) wasrecorded in the summer months May–August 1997and the narrowest (11–12 spp.) in February andMarch 1998.The highest monthly wind velocity wasregularly associated with higher fungal colonycounts than in case of the lowest velocity. Onthe other hand, wind direction did not exhibitany regular correlation either with the colonycounts of fungi or with the wind velocity. Highwind velocity could bring more fungal spores tobe sedimented on the surface of exposed agar.Diurnal fluctuations of fungal spores offungi displayed one peak at 12 noon when thehighest temperature and wind velocity, and theleast relative humidity were recorded and onetrough at midnight.     

Coordinate activation of human platelet protease-activated receptor-1 and -4 in response to subnanomolar alpha-thrombin

We previously demonstrated that human platelets activated with SFLLRN release PAR-1 activation peptide, PAR-1-(1-41), even in the presence of hirudin. This observation suggests that during their activation, platelets generate a protease that activates PAR-1. In this study, PAR-1 and -4 activation peptides were detected 10 s after 相似文献   

Coverage and uptake of systematic postal screening for genital Chlamydia trachomatis and prevalence of infection in the United Kingdom general population: cross sectional study

Objective To measure the coverage and uptake of systematic postal screening for genital Chlamydia trachomatis and the prevalence of infection in the general population in the United Kingdom. To investigate factors associated with these measures.Design Cross sectional survey of people randomly selected from general practice registers. Invitation to provide a specimen collected at home.Setting England.Participants 19 773 men and women aged 16-39 years invited to participate in screening.Main outcome measures Coverage and uptake of screening; prevalence of chlamydia.Results Coverage of chlamydia screening was 73% and was lower in areas with a higher proportion of non-white residents. Uptake in 16-24 year olds was 31.5% and was lower in men, younger adults, and practices in disadvantaged areas. Overall prevalence of chlamydia was 2.8% (95%confidence interval 2.2% to 3.4%) in men and 3.6% (3.1% to 4.9%) in women, but it was higher in people younger than 25 years (men 5.1%; 4.0% to 6.3%; women 6.2%; 5.2% to 7.8%). Prevalence was higher in the subgroup of younger women who were harder to engage in screening. The strongest determinant of chlamydial infection was having one or more new sexual partners in the past year.Conclusions Postal chlamydia screening was feasible, but coverage was incomplete and uptake was modest. Lower coverage of postal screening in areas with more non-white residents along with poorer uptake in more deprived areas and among women at higher risk of infection could mean that screening leads to wider inequalities in sexual health.     

Functional Overexpression and Purification of a Codon Optimized Synthetic Glucarpidase (Carboxypeptidase G2) in Escherichia coli

Glucarpidase (former name: carboxypeptidase G2, or CPG2) is a bacterial enzyme that is widely used in detoxification of the cytotoxic drug, methotrexate, and in Antibody Directed Enzyme Prodrug Therapy for cancer treatment. The glucarpidase gene of Pseudomonas sp. strain RS-16 was previously cloned in E coli, but expresses at a level that is approximately 100-fold lower than in the native strain. In this study, a synthetic gene coding for glucarpidase was codon-optimised and synthesized for maximum expression in E. coli using the vector pET28a. Our work indicated that the enzyme was expressed to ~60% of the total host protein and that purification of the recombinant His-tagged protein could be achieved in a single step by Ni²⁺ charged column chromatography. The synthetic recombinant glucarpidase expressed within this system was biologically active and zinc dependant. Our study showed that Mg²⁺ as well as Mn²⁺ ions inhibit the activity of the recombinant enzyme.