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Galbokka Hewage Roshanthi Eranga Karunaratne, Sandhya Sulochana Wijesundera, Dhammika Vidanagama, Chamila Priyangani Adikaram, Jennifer Perera
The emergence and spread of drug-resistant tuberculosis (TB) pose a threat to TB control in Sri Lanka. Isoniazid (INH) is a key element of the first-line anti-TB treatment regimen. Resistance to INH is mainly associated with point mutations in katG, inhA, and ahpC genes. The objective of this study was to determine mutations of these three genes in INH-resistant Mycobacterium tuberculosis (MTb) strains in Sri Lanka. Complete nucleotide sequence of the three genes was amplified by polymerase chain reaction and subjected to DNA sequencing...
April 23, 2018: Microbial Drug Resistance: MDR: Mechanisms, Epidemiology, and Disease
Tao Hu, Ying-Ning Zheng, Meng-Jie Li, Wen-Bin Liang, Yaqin Chai, Ruo Yuan
In this work, a highly sensitive photoelectrochemical (PEC) assay was constructed based on a donor-acceptor (D-A)-type material poly{4,8-bis[5-(2-ethylhexyl) thiophen-2-yl]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl-alt-3-fluoro-2-[(2-ethylhexyl)-carbonyl]thieno[3,4-b]thiophene-4,6-diyl} (PTB7-Th) as photoactive material and polyaniline (PANI) in-situ deposited on the surface of PTB7-Th as signal enhancer. Initially, PTB7-Th that contains an electron-rich unit as donor and an electron-deficient unit as acceptor with the easy separation of electron-hole pairs and intermolecular electron transfer provided an excellent photocurrent response...
April 20, 2018: Analytical Chemistry
Esmat Mangoli, Ali Reza Talebi, Morteza Anvari, Fatemeh Taheri, Mahboobeh Vatanparast, Tahereh Rahiminia, Akram Hosseini
OBJECTIVE: Aim of this study was to evaluate the effects of vitamin C on sperm parameters, sperm chromatin quality and apoptosis resulted of vitrification in neat semen and prepared spermatozoa of normozoospermic samples. MATERIAL AND METHODS: Forty semen samples from normozoospermic men were included in this prospective study. Each sample was divided into five groups. Group I: control or fresh semen, group II: semen prepared by swim-up method and then vitrified, group III: neat semen was vitrified, group IV: vitamin C (600 μM) was added to prepared spermatozoa and then vitrified and group V: vitamin C (600 μM) was added to neat semen and then vitrified...
April 2018: Taiwanese Journal of Obstetrics & Gynecology
Tahereh Rahiminia, Ehsan Farashahi Yazd, Farzaneh Fesahat, Mohammad Reza Moein, Ali Mohammad Mirjalili, Ali Reza Talebi
Objective: To investigate sperm chromatin/DNA integrity, global DNA methylation, and DNMT mRNA transcription in men with oligoasthenoteratozoospermia (OAT) compared with normozoospermic men. Methods: Semen samples from 32 OAT patients who comprised the case group and 32 normozoospermic men who comprised the control group were isolated and purified using a standard gradient isolation procedure according to World Health Organization criteria. DNMT1 , DNMT3A , and DNMT3B transcripts were then compared between groups using real-time quantitative reverse-transcription polymerase chain reaction...
March 2018: Clinical and Experimental Reproductive Medicine
Maikel Boot, Susanna Commandeur, Amit K Subudhi, Meriem Bahira, Trever C Smith, Abdallah M Abdallah, Mae van Gemert, Joël Lelièvre, Lluís Ballell, Bree B Aldridge, Arnab Pain, Alexander Speer, Wilbert Bitter
Due to the rise of drug resistant forms of tuberculosis there is an urgent need for novel antibiotics to effectively combat these cases and shorten treatment regimens. Recently, drug screens using whole cell analyses have been shown to be successful. However, current high-throughput screens focus mostly on stricto sensu life-death screening that give little qualitative information. In doing so, promising compound scaffolds or non-optimized compounds that fail to reach inhibitory concentrations are missed. To accelerate early TB drug discovery, we performed RNA sequencing on Mycobacterium tuberculosis and Mycobacterium marinum to map the stress responses that follow upon exposure to sub-inhibitory concentrations of antibiotics with known targets: ciprofloxacin, ethambutol, isoniazid, streptomycin and rifampicin...
April 16, 2018: Antimicrobial Agents and Chemotherapy
Joonhong Park, So Youn Shin, Kyungjong Kim, Kuhn Park, Soyoung Shin, Chunhwa Ihm
BACKGROUND: We examined the feasibility of a full-length gene analysis for the drug resistance-related genes inhA, katG, rpoB, pncA, rpsL, embB, eis, and gyrA using ion semiconductor next-generation sequencing (NGS) and compared the results with those obtained from conventional phenotypic drug susceptibility testing (DST) in multidrug-resistant Mycobacterium tuberculosis (MDR-TB) isolates. METHODS: We extracted genomic DNA from 30 pure MDR-TB isolates with antibiotic susceptibility profiles confirmed by phenotypic DST for isoniazid (INH), rifampin (RIF), ethambutol (EMB), pyrazinamide (PZA), amikacin (AMK), kanamycin (KM), streptomycin (SM), and fluoroquinolones (FQs) including ofloxacin, moxifloxacin, and levofloxacin...
July 2018: Annals of Laboratory Medicine
Faranak Rezaei, Mehri Haeili, Abbasali Imani Fooladi, Mohammad Mehdi Feizabadi
Objectives: Development of molecular techniques for rapid detection of drug resistant tuberculosis allows for the prompt initiation of appropriate anti-TB treatment. We aimed to assess high-resolution melting (HRM) analysis for the detection of rpsL, rrs and embB mutations to identify streptomycin and ethambutol resistance in Mycobacterium tuberculosis. Material and Methods: A total of 76 clinical isolates of M. tuberculosis including 25 SM-R, 21 EB-R and 30 drug susceptible - determined by the proportion method of drug susceptibility testing (DST) - were analyzed by HRM analysis, and the results were confirmed using DNA sequencing...
December 2017: Mædica
R Nazari-Vanani, N Sattarahmady, H Yadegari, H Heli
Bacteria, parasites and viruses are found widely in the environment as potential pathogens, and can be the source of infections. Therefore, sensitive and rapid methods for identification of the pathogens are required to achieve a better quality of life. Enterococcus faecalis commonly colonizes and threatens human health. In the present study, we demonstrate the fabrication of a novel electrochemical DNA biosensor based on electrodeposited gold nanostructures as a transducer substrate combined with toluidine blue (TB) as a redox marker...
March 20, 2018: Colloids and Surfaces. B, Biointerfaces
Kyle Ellrott, Matthew H Bailey, Gordon Saksena, Kyle R Covington, Cyriac Kandoth, Chip Stewart, Julian Hess, Singer Ma, Kami E Chiotti, Michael McLellan, Heidi J Sofia, Carolyn Hutter, Gad Getz, David Wheeler, Li Ding
The Cancer Genome Atlas (TCGA) cancer genomics dataset includes over 10,000 tumor-normal exome pairs across 33 different cancer types, in total >400 TB of raw data files requiring analysis. Here we describe the Multi-Center Mutation Calling in Multiple Cancers project, our effort to generate a comprehensive encyclopedia of somatic mutation calls for the TCGA data to enable robust cross-tumor-type analyses. Our approach accounts for variance and batch effects introduced by the rapid advancement of DNA extraction, hybridization-capture, sequencing, and analysis methods over time...
March 28, 2018: Cell Systems
Xia Yang, Shufan Wang, Yue Wang, Yi He, Yaqin Chai, Ruo Yuan
In this work, one stimuli-responsive DNA microcapsule was designed to combine duplex-specific nuclease (DSN) amplification strategy and SERS technology for sensitive detection of MicroRNA 155 (miRNA155). Firstly, Toluidine blue (TB) as Raman dye and CaCO3 as core-templates co-precipitated to form TB@CaCO3 composite. Then, DNA networks were layer by layer (LbL) constructed with oligonucleotide layers crosslinked by Linker ssDNA L to lock TB@CaCO3 inside. In the presence of EDTA, the core CaCO3 would be dissolved to form TB loading DNA microcapsule...
March 29, 2018: ACS Applied Materials & Interfaces
Najmeh Ansari, Kiarash Ghazvini, Mohammad Ramezani, Mahin Shahdordizadeh, Rezvan Yazdian-Robati, Khalil Abnous, Seyed Mohammad Taghdisi
The Mycobacterium Ag85 complex is the major secretory protein of M. tuberculosis. It is a potential marker for early diagnosis of tuberculosis (TB). The authors have identified specific aptamers for Ag85A (FbpA) via protein SELEX using magnetic beads. After twelve rounds of selection, two aptamers (Apt8 and Apt22) were chosen from different groups, and their binding constants were determined by flow cytometry. Apt22 (labeled with Atto 647N) binds to FbpA with high affinity (Kd  = 63 nM) and specificity...
December 5, 2017: Mikrochimica Acta
Husain Poonawala, Surbhi Leekha, Sandra Medina-Moreno, Mala Filippell, J Kristie Johnson, Robert R Redfield, Kapil K Saharia
BACKGROUNDHospitalized patients with suspected tuberculosis (TB) are placed in airborne isolation until 3 sputum smear samples are negative for acid-fast bacilli (AFB). The Xpert MTB/RIF assay ("Xpert") nucleic acid amplification test (NAAT) to identify Mycobacterium tuberculosis DNA and resistance to rifampicin is superior to AFB sputum smear microscopy for the diagnosis of TB.OBJECTIVETo compare the performance of a single Xpert to AFB smear microscopy for time to airborne infection isolation (AII) discontinuation...
May 2018: Infection Control and Hospital Epidemiology
Amin A Seleem, Belal H M Hussein
The objective of this study was to synthesize Gibberellic lanthanide complex and evaluate its biological activity to reduce the Gibberellic acid toxicity on liver and kidney. The new bis (Gibberellic)-nitro-terbium(III) complex was characterized by different analytical methods: elemental analyses, Uv-vis, molar ratio, fluorescence, FT-IR, and TGA-DTA measurements. Thirty newborns were classified into three groups control, Gibberellic acid, and Terbium gibberellic acid complex. Livers and kidneys of studied groups proceed for general histology and immunohistochemical staining of Cyr61, cytochrome C, and TNFR2...
March 25, 2018: Chemical Biology & Drug Design
Benson R Kidenya, Stephen E Mshana, Daniel W Fitzgerald, Oksana Ocheretina
BACKGROUND: Drug resistant Tuberculosis (TB) is considered a global public health threat. Whole-genome sequencing (WGS) is a new technology for tuberculosis (TB) diagnostics and is capable of providing rapid drug resistance profiles and genotypes for epidemiologic surveillance. Therefore, we used WGS to determine genotypic drug resistance profiles and genetic diversity of drug resistant Mycobacterium tuberculosis isolates from Mwanza, North-western Tanzania. METHODS: A cross-sectional study was conducted at the Bugando Medical Center (BMC) from September 2014 to June 2015...
March 2018: Tuberculosis
Jale Moradi, Maryam Izad, Mina Tabrizi, Nader Mosavari, Behnaz Esmaeili, Mohammad Mehdi Feizabadi
One third of the world population are latently infected with Mycobacterium tuberculosis and are at the risk of reactivation of tuberculosis (TB). The most effective strategy for control of TB worldwide is the development of a vaccine that inhibits progression of latent TB to active infection. In this study, two optimized constructs consisting of multi-epitopes DNA derived from three latency antigens Rv2029c, Rv2031c, and Rv2627c fused with or without light chain 3 (LC3) are synthetized. The immunogenicity effectiveness of two DNA constructs was evaluated in the mouse model...
March 19, 2018: Acta Microbiologica et Immunologica Hungarica
Jana Amlerova, Ibrahim Bitar, Jaroslav Hrabak
Tuberculosis (TB) is considered one of the most serious infectious diseases worldwide. Effective control of tuberculosis infection involves multiple steps, such as reliable detection, treatment, an epidemiological control as a part of case management, and further surveillance and monitoring of TB spread in the human population. Due to the accelerating advances in molecular biology, especially in DNA sequencing, in the past decade, the application of these methods has become crucial for TB evolution studies, differentiation of Mycobacterium tuberculosis genotypes, and their distribution...
March 17, 2018: Folia Microbiologica
Maopeng Tian, Zijie Zhou, Songwei Tan, Xionglin Fan, Longmeng Li, Nadeem Ullah
Despite the vaccine Mycobacterium bovis Bacillus Calmette-Guérin is used worldwide, tuberculosis (TB) remains the first killer among infectious diseases. An effective vaccine is urgently required. DNA vaccine has shown prophylactic as well as therapeutic effects against TB, while its weak immunogenicity hinders the application. As a strong inducer of Th1-biased immune response, DMT, consisting of dimethyldioctadecylammonium (DDA) and two pattern recognition receptor agonists monophosphoryl lipid A and trehalose 6,6'-dibehenate (TDB), was a newly developed liposomal adjuvant...
2018: Frontiers in Immunology
Mehmet Demirci, Suat Saribas, Nigar Ozer, Sezer Toprak, Emel Caglar, Gonenc Ortakoylu, Pelin Yuksel, Gulsel Ayaz, Hrisi Bahar Tokman, Omer Uysal, Harika Oyku Dinc, Tevhide Ziver, Bekir Kocazeybek
BACKGROUND: Several nucleic acid amplification techniques (IS6110, 16S rRNA, and 85B mRNA) were developed for the rapid, direct detection of Mycobacterium tuberculosis. We aimed to assess the diagnostic performance of 85B mRNA-based RT-qPCR by comparing with the real-time PCR COBAS TaqMan MTB Kit while using the BACTEC MGIT 960 method as the gold standard. METHODS: 60 patients with confirmed pulmonary TB and 60 individuals without TB were included as the study and control groups, respectively...
March 8, 2018: Journal of Infection and Public Health
Astha Giri, Shraddha Gupta, Hassan Safi, Anshika Narang, Kamal Shrivastava, Naresh Kumar Sharma, Subramanya Lingaraju, Mahmud Hanif, Anuj Bhatnagar, Balakrishnan Menon, David Alland, Mandira Varma-Basil
Mutations at embB306 are the most prevalent polymorphisms associated with ethambutol (EMB) resistance, responsible for 40-60% of EMB resistant clinical cases of tuberculosis (TB). The present study analyzed additional mutations associated with EMB resistance in the embB, embC, embA and Rv3806c (ubiA) genes in 29 EMB resistant and 29 EMB susceptible clinical isolates of M. tuberculosis selected from 360 patients with TB. The entire ubiA gene, mutational hotspot regions of embB, embC, and upstream region of embA were screened for polymorphisms by DNA sequencing and the results correlated with minimum inhibitory concentrations (MIC) of EMB...
January 2018: Tuberculosis
Nastassja L Kriel, James Gallant, Niël van Wyk, Paul van Helden, Samantha L Sampson, Robin M Warren, Monique J Williams
Nucleoid associated proteins (NAPs) are known organisers of chromosomal structure and regulators of transcriptional expression. The number of proposed NAPs in mycobacteria are significantly lower than the number identified in other organisms. An interesting feature of mycobacterial NAPs is their low sequence similarity with those in other species, a property that has hindered their identification. In this review, we discuss the current evidence for the proposed classification of six mycobacterial proteins, Lsr2, EspR, mIHF, HupB, MDP2 and NapM, as NAPs in mycobacterial species with an emphasis on their roles in modulating chromosome structure and transcriptional regulation...
January 2018: Tuberculosis
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