In-silico metabolome target analysis towards PanC-based antimycobacterial agent discovery

Mycobacterium tuberculosis, the main cause of tuberculosis [TB], has still remained a global health crisis especially in developing countries. Tuberculosis treatment is a laborious and lengthy process with high risk of noncompliance, cytotoxicity adverse events and drug resistance in patient. Recently, there has been an alarming rise of drug resistant in TB. In this regard, it is an unmet need to develop novel antitubercular medicines that target new or more effective biochemical pathways to prevent drug resistant Mycobacterium. Integrated study of metabolic pathways through in-silico approach played a key role in antimycobacterial design process in this study. Our results suggest that pantothenate synthetase [PanC], anthranilate phosphoribosyl transferase [TrpD] and 3-isopropylmalate dehydratase [LeuD] might be appropriate drug targets. In the next step, in-silico ligand analysis was used for more detailed study of chemical tractability of targets. This was helpful to identify pantothenate synthetase [PanC, Rv3602c] as the best target for antimycobacterial design procedure. Virtual library screening on the best ligand of PanC was then performed for inhibitory ligand design. At the end, five chemical intermediates showed significant inhibition of Mycobacterium bovis with good selectivity indices [SI] >/= 10 according to Tuberculosis Antimicrobial Acquisition and Coordinating Facility of US criteria for antimycobacterial screening programs

Cellular immunity survey against urinary tract infection using pVAX/fimH cassette with mammalian and wild type codon usage as a DNA vaccine

PURPOSE: FimH (the adhesion fragment of type 1 fimbriae) is implicated in uropathogenic Escherichia coli (UPEC) attachment to epithelial cells through interaction with mannose. Recently, some studies have found that UPEC can thrive intracellularly causing recurrent urinary tract infection (UTI). Almost all vaccines have been designed to induce antibodies against UPEC. Yet, the humoral immune response is not potent enough to overcome neither the primary UTI nor recurrent infections. However, DNA vaccines offer the possibility of inducing cell mediated immune responses and may be a promising preventive tool. MATERIALS AND METHODS: In this study, we employed two different open reading frames within mammalian (mam) and wild type (wt) codons of fimH gene. Optimized fragments were cloned in pVAX-1. Expression of the protein in COS-7 was confirmed by western blot analysis after assessing pVAX/fimH(mam) and pVAX/fimH(wt). The constructs were injected to BALB/c mice at plantar surface of feet followed by electroporation. RESULTS: The mice immunized with both constructs following booster injection with recombinant FimH showed increased interferon-gamma and interleukin-12 responses significantly higher than non-immunized ones (p<0.05). The immunized mice were challenged with UPEC and then the number of bacteria recovered from the immunized mice was compared with the non-immunized ones. Decreased colony count in immunized mice along with cytokine responses confirmed the promising immune response by the DNA vaccines developed in this study. CONCLUSION: In conclusion, DNA vaccines of UPEC proteins may confer some levels of protection which can be improved by multiple constructs or boosters.

effects of altered ultrasound parameters on the recovery of sciatic nerve injury

Initial studies have shown that low-energy ultrasound stimulates living tissue cells to reduce regeneration or speed up their recovery. The purpose of this study was to examine the effects of various ultrasound parameters on the speed of recovery in injured sciatic nerves. NMRI mice [n = 200] with injured left paw, caused by crushing their sciatic nerves, were randomly selected. The animals were exposed to ultrasound radiation with various frequencies, intensities, and exposure time. They were allocated into 20 groups [19 treatment and 1 control groups]. Sciatic functional index [SFI] test was used to evaluate the difference between the groups with respect to functional efficiency of the sciatic nerve and its recovery. The results of SFI test obtained from the 14th day showed a significant difference among the groups [P<0.05]. On the 14th day after treatment, one of the groups [US11] recovered up to 90%. Altered ultrasound exposure parameters had more favorable outcomes compared with our previous work