Modulation of NKG2D Expression in Human CD8+ T Cells Corresponding with Tuberculosis Drug Cure

Background

Biomarkers predicting tuberculosis treatment response and cure would facilitate drug development. This study investigated expression patterns of the co-stimulation molecule NKG2D in human tuberculosis and treatment to determine its potential usefulness as a host biomarker of tuberculosis drug efficacy.

Methods

Tuberculosis patients (n = 26) were recruited in Lahore, Pakistan, at diagnosis and followed up during treatment. Household contacts (n = 24) were also recruited. NKG2D expression was measured by qRT-PCR in RNA samples both ex vivo and following overnight mycobacterial stimulation in vitro. Protein expression of NKG2D and granzyme B was measured by flow cytometry.

Results

NKG2D expression in newly diagnosed tuberculosis patients was similar to household contacts in ex vivo RNA, but was higher following in vitro stimulation. The NKG2D expression was dramatically reduced by intensive phase chemotherapy, in both ex vivo blood RNA and CD8⁺ T cell protein expression, but then reverted to higher levels after the continuation phase in successfully treated patients.

Conclusion

The changes in NKG2D expression through successful treatment reflect modulation of the peripheral cytotoxic T cell response. This likely reflects firstly in vivo stimulation by live Mycobacterium tuberculosis, followed by the response to dead bacilli, antigen-release and finally immunopathology resolution. Such changes in host peripheral gene expression, alongside clinical and microbiological indices, could be developed into a biosignature of tuberculosis drug-induced cure to be used in future clinical trials.     

Mycosubtilin overproduction by Bacillus subtilis BBG100 enhances the organism's antagonistic and biocontrol activities

A Bacillus subtilis derivative was obtained from strain ATCC 6633 by replacement of the native promoter of the mycosubtilin operon by a constitutive promoter originating from the replication gene repU of the Staphylococcus aureus plasmid pUB110. The recombinant strain, designated BBG100, produced up to 15-fold more mycosubtilin than the wild type produced. The overproducing phenotype was related to enhancement of the antagonistic activities against several yeasts and pathogenic fungi. Hemolytic activities were also clearly increased in the modified strain. Mass spectrometry analyses of enriched mycosubtilin extracts showed similar patterns of lipopeptides for BBG100 and the wild type. Interestingly, these analyses also revealed a new form of mycosubtilin which was more easily detected in the BBG100 sample. When tested for its biocontrol potential, wild-type strain ATCC 6633 was almost ineffective for reducing a Pythium infection of tomato seedlings. However, treatment of seeds with the BBG100 overproducing strain resulted in a marked increase in the germination rate of seeds. This protective effect afforded by mycosubtilin overproduction was also visualized by the significantly greater fresh weight of emerging seedlings treated with BBG100 compared to controls or seedlings inoculated with the wild-type strain.     

The response of transgenic <Emphasis Type="Italic">Brassica</Emphasis> species to salt stress: a review

Salt stress is considered one of the main abiotic factors to limit crop growth and productivity by affecting morpho-physiological and biochemical processes. Genetically, a number of salt tolerant Brassica varieties have been developed and introduced, but breeding of such varieties is time consuming. Therefore, current focus is on transgenic technology, which plays an important role in the development of salt tolerant varieties. Various salt tolerant genes have been characterized and incorporated into Brassica. Therefore, such genetic transformation of Brassica species is a significant step for improvement of crops, as well as conferring salt stress resistance qualities to Brassica species. Complete genome sequencing has made the task of genetically transforming Brassica species easier, by identifying desired candidate genes. The present review discusses relevant information about the principles which should be employed to develop transgenic Brassica species, and also will recommend tools for improved tolerance to salinity.     

Effect of growing region on quality characteristics and phenolic compounds of chemlali extra-virgin olive oils

The present work aims at the characterizing chemlali extra-virgin olive oils from different locations in northern, central and southern Tunisia in terms of their quality indices, fatty acids, sterol content, phenolic composition and sensory profiles to show the classification of oil samples according to the geographical area. The majority of the analytical parameters have presented statistically significant differences (p < 0.05). The main sterols found in all chemlali olive oils were β-sitosterol, ?-5-avenasterol, campesterol and stigmasterol. The phenolic compounds present in five olive oil samples were analysed by high-performance liquid chromatography (HPLC) method, thus identifying 16 phenolic compounds belonging to different phenolic types. The results have shown no qualitative differences in the phenolic fractions among extra-virgin olive oils from different geographical regions. However, the quantitative differences were observed in a wide number of phenolic compounds. In all studied olive oil samples, secoiridoids were the most abundant, followed by lignans, phenolic alcohols and flavonoids, respectively. Although there is no significant influence on the sensory scores of oils, some slight changes in sensorial profiles were noted: slightly higher intensities of sensory characteristics that are pungent, fruity and bitter in chemlali olive oil from Hammamet and Gafsa.     

Fluorescence probe enhanced spectrofluorimetric method for the determination of sparfloxacin in tablets and biological fluids

Yttrium‐sensitized fluorescence was used to develop a sensitive and simple spectrofluorimetric method for the determination of sparfloxacin. The method is based on the strong fluorescence of sparfloxacin after adding the fluorescence probe yttrium in buffer solution (pH = 8), and various factors influencing fluorescence were investigated. Under optimum conditions, the enhanced fluorescence intensity of the system showed a good linear relationship with the concentration of sparfloxacin over the range 8 × 10^?7 to 1.4 × 10^?5 mol L^?1 with a correlation coefficient of 0.9997. The detection limit (S/N = 3) was determined as 9.01 × 10^?8 mol L^?1. The mechanism of the sensitizing effect of probe was discussed. This method has been successfully applied for the determination of sparfloxacin in pharmaceuticals, human urine and serum samples; the result obtained was satisfactory. Copyright © 2010 John Wiley & Sons, Ltd.     

Toxic potential of some indigenous plant oils against the rice weevil,Sitophilus oryzae (Linnaeus)

The present study was conducted to evaluate the toxic potential of five indigenous plant oils: black pepper (Piper nigrum), Chinese cinnamon (Cinnamomum cassia), garlic (Allium sativum), river red gum (Eucalyptus camaldulensis), and yellow oleander (Thevetia peruviana), against laboratory reared Sitophilus oryzae adults. The bioassays were done by the diet incorporation method with concentrations ranging from 50 ppm to 500 ppm. Based on lethal concentrations to kill 50% (LC₅₀) of the subjected weevils, T. peruviana proved to be the most toxic having the lowest LC₅₀ values, 414.58, 201.94, and 129.52 ppm, after 7, 14, and 21 days of exposure, respectively, followed by E. camaldulensis (475.51, 366.65, and 251.28 ppm, respectively). The rest of the plant oils also showed toxic potential, but these were less toxic compared with T. peruviana and E. camaldulensis. With respect to the time taken to cause 50% mortality (LT₅₀) of the exposed weevils, T. peruviana had LT₅₀ at 14.54 days followed by P. nigrum (22.09 days), E. camaldulensis (24.29 days), and C. cassia (28.71 days). Whereas, A. sativum took the longest time (44.47 days) to cause 50% mortality of the exposed weevils. In conclusion, the result revealed toxic potential of tested plant oils, and suggests further studies under simulated‐field conditions should be included in the management plan for S. oryzae.     

CD80/CD86 costimulation regulates acute vascular rejection

Xenotransplantation may provide the only solution to the shortage of human donor organs. Although hyperacute rejection associated with xenotransplantation can now be overcome, acute vascular rejection (AVR) remains a primary barrier to xenotransplantation. To date, standard immunosuppressive agents fail to block AVR or prolong xenograft survival. The present study was undertaken to determine the role of CD80/CD86 costimulatory molecules in regulating AVR. Lewis rat hearts were transplanted heterotopically into wild-type or IL-12, CD80- or CD86-deficient C57BL/6 mice. Wild-type recipients were treated with CD80 or CD86 neutralizing Ab with and without daily cyclosporin A (CsA, 15 mg/kg). Transplanted hearts in untreated wild-type recipients were rejected on postoperative days (POD) 17-21 and showed cell-mediated rejection (CMR) and AVR pathologies. In contrast, transplanted hearts in IL-12 and CD80 recipients or wild-type recipients treated with CD80 neutralizing Ab were rapidly rejected on POD 5 and 6 with AVR pathology. Interestingly, hearts transplanted into CD86 knockout recipients or wild-type recipients treated with CD86 neutralizing Ab underwent CMR on POD 17. Finally, blockade of CD86 but not CD80 rendered xenograft recipients sensitive to daily CsA therapy, leading to indefinite xenograft survival. To conclude, we demonstrate that AVR can be overcome by blocking the CD86 costimulatory pathway. Furthermore, we demonstrate that CD80 and CD86 have opposing roles in regulation of xenotransplantation rejection, where CD80 drives CMR and attenuates AVR while CD86 drives AVR. Most strikingly, indefinite xenograft survival can be achieved by suppressing AVR with CD86 neutralization in combination of CsA therapy, which inhibits CMR.     

Identification of nerve plexi in connective tissues of the sea cucumber Holothuria glaberrima by using a novel nerve-specific antibody

The echinoderm nervous system is one of the least studied among invertebrates, partly because the tools available to study the neurobiology of this phylum are limited. We have now produced a monoclonal antibody (RN1) that labels a nervous system component of the sea cucumber Holothuria glaberrima. Western blots show that our antibody recognizes a major band of 66 kDa and a minor band of 53 kDa. Immunohistological experiments show that, in H. glaberrima, the antibody distinctly labels most of the known nervous system structures and some components that were previously unknown or little studied. A surprising finding was the labeling of nervous plexi within the connective tissue compartments of all organs studied. Double labeling with holothurian neuropeptides and an echinoderm synaptotagmin showed that RN1 labeled most, if not all, of the fibers labeled by these neuronal markers, but also a larger component of cells and fibers. The presence of a distinct connective tissue plexus in holothurians is highly significant since these organisms possess mutable connective tissues that change viscosity under the control of the nervous system. Therefore, the cells and fibers recognized by our monoclonal antibodies may be involved in controlling tensility changes in echinoderm connective tissue.     

The Study of Chromium and Zinc Contaminated Soil Influence on Iron Content and Protein Profile of Ornamental Cabbage Plant

Biology Bulletin - To evaluate the accumulation potential of zinc (Zn) and chromium (Cr) by Brassica oleracea L. plant, 15-day-old seedlings were exposed to different concentrations of Zn and Cr...