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   Table of Contents - Current issue
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January-March 2019
Volume 3 | Issue 1
Page Nos. 1-66

Online since Wednesday, March 13, 2019

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ORIGINAL ARTICLES  

High-resolution genotyping of Mycobacterium tuberculosis isolates from syria using mycobacterial interspersed repetitive unit-variable-number tandem repeat Highly accessed article p. 1
Hana Zarzour, Ammar Madania, Ifad Ghoury, Maya Habous
DOI:10.4103/bbrj.bbrj_139_18  
Background: We aimed to evaluate the utility of 24 loci mycobacterial interspersed repetitive unit-variable-number tandem repeat (MIRU-VNTR) genotyping method for discrimination of clinical Mycobacterium tuberculosis isolates in Syria. Methods: We studied 68 clinical tuberculosis (TB) isolates originating from unrelated Syrian TB patients from different regions in Syria. Genetic types (consisting of 24 digits) were determined and used to construct a dendrogram. Results: Fifty-six distinct MIRU patterns were revealed, from which 52 patterns were represented by unique isolates. Sixteen isolates were distributed into 4 clusters, 3 of which consisted of isolates belonging to the TUR lineage. Nine MIRUs showed high Hunter-Gaston index (HGI) values (>0.6), with QUB-26 having the highest discriminatory power (HGI = 0.821), followed by MIRU10, MIRU26, MIRU16, and Mtub39. The cumulative HGI value of the 24-MIRU set was 0.985. Interestingly, using the reduced 15-MIRU, set resulted in the same HGI. The TUR lineage was the most frequent in our sample (23.5%), and it appears that it is widespread in Syria as in Turkey. The discriminatory power of MIRU-VNTR among the subset belonging to the TUR lineage was extremely low due to the high clustering rate (62.5%) of TUR isolates, indicating that this method is inappropriate to discriminate isolates of this lineage. On the other hand, the Beijing lineage was not represented in our isolates. Conclusion: We demonstrated the high prevalence of the TUR lineage and the low prevalence of the Beijing lineage among Syrian clinical TB isolates. The MIRU-VNTR method was highly discriminative among non-TUR TB isolates, but it was inappropriate to discriminate isolates of the TUR lineage.
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Comparative genomics of Mycoplasma: Insights on genome reduction and identification of potential antibacterial targets p. 9
Angamuthu Kandavelmani, Shanmughavel Piramanayagam
DOI:10.4103/bbrj.bbrj_142_18  
Background: Mycoplasmas are cell wall-deficient bacteria which cause respiratory and urogenital infections in human. Mycoplasmas are resistant to many of the available antibiotics because of their high mutation rates and lack of cell wall. Various studies have identified the emergence of treatment-resistant Mycoplasma isolates. Novel drug target identification has become significant for the development of successful antibacterial treatments. Computational genomic analysis plays a significant role in facilitating the identification of potential drug targets in many bacterial pathogens. Methods: In the present study, 12 Mycoplasma genomes were subjected to comparative genomic analysis to reinforce the understanding of their genomic organization and to identify potential drug targets. The distributions of genes under the Clusters of Orthologous Groups of proteins (COG) functional categories were analyzed for all the 12 Mycoplasma genomes. Genes from each functional category that are conserved across all the Mycoplasma genomes were extracted to identify the backbone genome of Mycoplasma species. The genes in the backbone genome were subjected to similarity search against a database of essential genes to validate their essentiality. Essentiality of these genes was further analyzed based on their function and subcellular localization. Results: The 12 Mycoplasma genomes under study were found to exhibit marked similarities in COG functional category distributions. An overall loss of genes in various functional categories has been observed in all the 12 Mycoplasma genomes. In all the 12 Mycoplasma genomes under study, a maximum reduction in the genes involved in the secondary metabolites biosynthesis, transport, and catabolism (Q) is observed. Conclusion: Comparative genomic studies have identified a set of 170 genes which are commonly present all the 12 Mycoplasma genomes. Further analysis of these genes has identified a set of 158 core essential genes which serve as a promising cluster of novel antibacterial targets.
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Tuberculous pleural effusion: Modification of metabolome by the effect of common metabolic disease, diabetes mellitus p. 19
Pathum Sookaromdee, Viroj Wiwanitkit
DOI:10.4103/bbrj.bbrj_34_19  
Background: Pleural effusion is an important problem in chest medicine. This problem might be seen in several conditions including tuberculosis and cancer. The differential diagnosis between tuberculous effusion and other effusions usually requires a good diagnostic test. The advance metabolomics approach is mentioned for the usefulness for differential diagnosis. The important consideration is on the effect of background metabolic alterations of the patients. Methods: The authors hereby assess the effect of background diabetes mellitus on the metabolome in tuberculous effusion. The common specific metabolome between tuberculous effusion and diabetes is searched for. The simulation to assess the confounding and overlapping effect of metabolome in diabetes to tuberculous effusion is done. Results: The only one identified common metabolome is citric acid. According to the simulation, there is a significant confounding and overlapping effect of citric acid in diabetes to tuberculous effusion. Conclusion: Diabetes might interfere with the use of citric acid as a metabolomic biomarker for tuberculous effusion.
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Establishing reference ranges and normal values for coagulation screening in healthy Indian male volunteers enrolled for a longitudinal study p. 22
Rehan Ahmed, Prosenjit Ganguli, Natwar Singh, Surinderpal Singh, Umesh Das Gupta, Yogesh K Jaiswal, Zahid Asharaf, Prasanna Reddy, Velu Nair
DOI:10.4103/bbrj.bbrj_140_18  
Background: The study was designed for establishing reference ranges and normal values for coagulation screening in healthy Indian male volunteers. At present, there are no standard parameters established for coagulation screening assays of Indian population. The parameters used as a reference in the coagulation assays are of Western origin. We know that ethnicity of the western population is different from the Indian population which may result in a different set of reference ranges of the coagulation assay. It is necessary to determine the mean normal values for the coagulation assay, namely prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen in an Indian population. The aim of the study was to establish the reference ranges of coagulation assays for all future experiments on the cohort. Methods: Six hundred healthy male volunteers were sampled and underwent coagulation testing at a seven hundred-bedded hospital at Jammu (Jammu). Fresh normal pool plasma (FNPP) was prepared concurrently and investigated (n = 50 volunteers). In the study population, the arithmetic means of the coagulation assay were as follows: PT: 13.00 s, APTT: 34 s, TT: 17.3 s, and fibrinogen: 298 mg/dl (19 s), and in the FNPP, it was 12.8 s, 33.2 s, 17 s, and 298 mg/dl (19 s), respectively. Results: The reference range of coagulation screening in our study was established as the following values: PT: 10.7–15.3 s, APTT: 26.8–41.2 s, TT: 12.8–21.7 s, and fibrinogen: 223–372 mg/dl. This study has set a reference range of normal values for coagulation assay screening in longitudinal studies where these tests were repeated on the same set of individuals at six-month interval for the subsequent 3 years. Conclusion: We see no effect of age on Coagulation cascade in our study. Overall mean values resembled with various age groups in coagulation cascade. These parameters of Coagulation cascade set a standard for high altitude studies where these tests are in normal procedure.
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In vitro antimicrobial activity of aqueous extracts of Ocimum suave Willd., Plectranthus barbatus andrews and Zanthoxylum chalybeum Engl. against selected pathogenic bacteria p. 30
Joseph Mwanzia Nguta, Micheni Ndei Kiraithe
DOI:10.4103/bbrj.bbrj_128_18  
Background: There is an urgent need for new antimicrobials, due to the increase in drug resistance. Current studies suggest that, by 2050, in the absence of major improvements in drug discovery, more individuals will die from drug-resistant bacterial infections than from cancer. This will result in a cumulative effect on the global gross domestic product of as much as 100 trillion dollars. To discover new drugs, new targets, and lead compounds are badly needed. Natural products of plant biodiversity are a key source of starting points for novel antimicrobial agents with activity against sensitive and resistant bacterial strains. The current study is timely and of a high impact, since it sought to validate anecdotal efficacy of aqueous extracts from selected medicinal plants conventionally used against microbial infections, namely, leaves from Ocimum suave Willd. (Lamiaceae), roots from Plectranthus barbatus Andrews (Lamiaceae), and roots from Zanthoxylum chalybeum Engl. (Rutaceae). Methods: The study plant parts were collected from Msambweni Sub-county, Kwale County, Kenya. Agar well diffusion method was used to evaluate the antimicrobial activity of the extracts against methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 1385), Escherichia coli (ATCC 25922), S. aureus (ATCC 25923), and Bacillus cereus (ATCC 11778). Results: Root extract of P. barbatus was found to be the most active of the studied extracts exhibiting mean inhibition zone values of 18.67 mm, 20.00 mm, and 25.33 mm in S. aureus, MRSA and B. cereus strains, respectively, after 24 h of incubation. All the studied extracts did not exhibit activity against E. coli. In addition, the aqueous leaf extract from the leaves of O. suave did not display antimicrobial activity against the tested organisms. Conclusion: These findings justify the continued ethno pharmacological utilization of P. barbatus and Z. chalybeum extracts against bacterial infections in traditional herbal medicine by various local communities. Furthermore, the current findings lay a strong foundation for further investigation of extracts from P. barbatus and Z. chalybeum for isolation, identification, and characterization of bioactive molecules responsible for the observed antimicrobial activity. These molecules could serve as templates for the discovery of a new class of antimicrobial agents for the management of economically important bacterial infections.
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Biosecurity-related biomedical laboratory practices in Pakistan: Educating personnel regarding transportation of biohazardous substances p. 35
Humaira Shafaq, Shahida M Qureshi, Sadia Shakoor
DOI:10.4103/bbrj.bbrj_20_19  
Background: Unchecked and unregulated transport of biohazardous material is a serious risk to the population and the environment. More importantly, packaging and knowledge of transport requirements and regulations is essential among personnel preparing and dispatching biohazardous shipments. To ensure safe transport practices among biomedical staff, regular training sessions are required but are not frequently carried out. We report on pre- and posttest knowledge among clinical laboratory and biomedical staff from southern Pakistan after such training. Methods: A basic questionnaire developed from the International Air Transport Association Dangerous Goods Regulations was prepared and administered to 36 participants in two different workshops carried out in April 2016 in Karachi, Pakistan. Questionnaires were administered before (pretest) and after (posttest) the practical training exercises to test comprehension. Scores from each participant were entered in MS Excel. Normalized learning gains (NLGs) were calculated for each participant. Results: Participants demonstrated lack of baseline knowledge of transport regulations and procedures for biohazardous material. After a full-day training session comprising lectures, demonstrations, as well as hands-on application of triple packaging requirements for refrigerated, dry ice, and ambient temperature shipments, the average NLG for 36 participants was 0.7 ± 0.3. Conclusions: Our results show that baseline knowledge of biosecurity and transport guidelines among laboratory personnel in southern Pakistan is poor. Moreover, hands-on workshops can be instrumental in increasing knowledge and understanding of these guidelines among personnel and should be conducted regularly.
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Effect of ATP-Binding cassette, Sub-Family B polymorphism on plasma nevirapine and efavirenz levels in human immunodeficiency virus with tuberculosis-infected cases: A molecular structure analysis p. 39
Pathum Sookaromdee, Viroj Wiwanitkit
DOI:10.4103/bbrj.bbrj_23_19  
Background: Tuberculosis (TB) is an important infection that is usually seen in the patient with underlying human immunodeficiency virus (HIV) infection. An important concern on the cases with HIV/TB is the management of HIV by antiretroviral drug treatment (antiretroviral therapy). The effect of underlying genetic factor is interesting. The effect of adenosine triphosphate-binding cassette, sub-family B (ABCB-1) polymorphism on plasma nevirapine and efavirenz levels is inconclusive. Methods: The authors perform standard molecular structure analysis to assess the effect of C and T alleles of ABCB-1 C3435T polymorphism. The phenotype expressions, reflected by nevirapine and efavirenz levels, are comparatively studied. Results: With focus on 3435 position, the ratio of derived molecular weight after quantum assessment, comparing CC to CT and TT genotypes, is equal to 1:1.068:1.135. Conclusion: The authors can identify the effect of ABCB-1 on the plasma nevirapine and efavirenz levels in HIV/TB patients.
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Rate of children at Thailand–Myanmar border area prone to tuberculosis infection: An epidemiological prediction with referencing to bacillus Calmette–Guerin vaccination in Thailand p. 42
Beuy Joob, Viroj Wiwanitkit
DOI:10.4103/bbrj.bbrj_21_19  
Background: The high incidence of tuberculosis exists in several areas around the world. In Indochina, the area between Thailand and Myanmar is presently the problematic endemic area of tuberculosis. The extremely high incidence of tuberculosis at this area becomes an important global public health consideration. Methods: In this specific article, the authors perform a mathematical model study based on the local available data on the efficacy and coverage rate of Bacillus Calmette–Guerin vaccination in Thailand. The specific situation at the Thailand–Myanmar border is focused in the present assessment. Results: The high incidence of children at Thailand–Myanmar border area prone to tuberculosis infection can be detected. Conclusion: The increase Bacillus Calmette–Guerin coverage and finding for new effective tuberculosis vaccine might be necessary for reduce the problem of high incidence of tuberculosis at the Thailand and Myanmar border area.
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Pulmonary tuberculosis incidence and drug resistance-related factors analysis in North China p. 46
Zheng Qi, Wei Yang, Yan-Fu Wang, Chang Liu
DOI:10.4103/bbrj.bbrj_141_18  
Background: This study analysis the epidemiological characteristics of incidence, gender, age, occupation, educational level, income, residence location, and timescale of 773 pulmonary tuberculosis patients in Hulin County in North China from 2014 to 2016. Methods: Two hundred and sixty-seven strains were identified and nine drug sensitivity tests against isoniazid (INH), streptomycin (S), ethambutol (EMB), rifampicin (RFP), ofloxacin (OFX), capreomycin, prothionamide, para-aminosalicylic acid (PAS), and kanamycin. Results: The top five drug resistance sequences of nine kinds of anti-TB drugs were S (24.535%) >INH (13.383%) >RFP (8.178%) >OFX (4.089%) >EMB (1.859%). Conclusion: The anti-TB medicine needs to be use under the guidelines to minimize drug resistance, and the elder age and poor also the work focus in the future.
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Evaluation of pulmonary hypertension in stable chronic obstructive pulmonary disease patients using transthoracic echocardiography p. 53
Swetabh Purohit, Arsh Garg, Vinod Joshi, Gopal Purohit, Varsha Joshee
DOI:10.4103/bbrj.bbrj_4_19  
Background: Pulmonary hypertension (PH) frequently complicates chronic obstructive pulmonary disease (COPD) process and adversely affects the exercise capacity of such patients. However, majority of the COPD patients are not routinely evaluated for PH even though a rapid screening tool in the form of transthoracic echocardiography (TTE) is available for the early diagnosis of PH. This study was conducted with an aim to study the frequency and severity of PH among stable COPD patients. Materials and Methods: Fifty stable COPD patients who met the inclusion criteria were selected and staged for severity by spirometry and screened for PH using TTE. Tricuspid regurgitation jet velocity obtained was used to calculate systolic pulmonary artery pressure (sPAP). Mean pulmonary artery pressure was calculated from sPAP using Chemla formula. Results: (1) The frequency of PH in COPD patients was 48% (24 out of 50). The frequency of PH in patients of mild, moderate, severe, and very severe GOLD COPD stages was 12.5% (1/8), 45.83% (11/24), 61.53% (8/13), and 80% (4/5), respectively. (2) Out of the 24 patients with PH, the relative incidence of mild (25–35 mmHg), moderate (35–45 mmHg), and severe (>45 mmHg) PH was 54.16% (13/24), 29.16% (7/24), and 16.67% (4/24), respectively. Only one patient had “out-of-proportion” PH. Conclusion: PH is an underdiagnosed entity that frequently complicates the course of disease in COPD patients and adversely affects exercise capacity. TTE is an accurate, noninvasive screening tool for PH and should be routinely used in all COPD patients for early diagnosis of PH.
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Histological and biochemical changes induced by ethanolic leaf extract of Moringa oleifera in the liver and lungs of adult wistar rats p. 57
Ferdinand Uwaifo, Anthony Ngokere, Ejatuluchukwu Obi, Matthew Olaniyan, Oladeinde Bankole
DOI:10.4103/bbrj.bbrj_126_18  
Background: Moringa oleifera, otherwise referred to as a miracle tree, has been used in the treatment of numerous diseases. Moringa has been the subject of intensive scientific research; however, there has been scant information on its sub-acute effect on the kidneys. This work was done to investigate the histological and biochemical effect of M. oleifera on the liver and lungs of adult Wistar rats. Twenty-eight adult healthy rats were used for this study. The rats were divided into four groups of seven per group and fed with pellets and water ad libitum. Group A served as the control, Group B was fed with 500 mg/kg of ethanolic extract of Moringa leaf, Group C with 1000 mg/kg, while Group D with 1500 mg/kg body weight. Methods: Doses were administered once daily using oral gavage for 28 days. Before the time of sacrificing the animals, blood samples were collected into plain test tubes and the animals anesthetized before sacrifice. Necropsy was performed and the tissues (liver and lungs) processed and stained using hematoxylin and eosin staining techniques. Some liver enzymes (aspartate aminotransferase [AST], alanine aminotransferase, alkaline phosphatase) and albumin were analyzed. Results: There were significant increases in AST and albumin in the 1000 mg/kg (19.50 ± 2.32, P = 0.036 and39.18 ± 8.56, P = 0.041) and 1500 mg/kg (26.55 ± 3.81, P = 0.021 and 46.75 ± 8.57, P = 0.021), respectively, in the treated animals when compared with the control (21.47 ± 16.33 and 4.54 ± 1.09). Histoarchitecture of the liver showed vascular congestion in the 1500 mg/kg treated rats which is indicative of congestive hepatopathy. There was no noticeable structural damage on the lungs. Conclusion: The result showed that M. oleifera may be toxic to the liver at concentrations higher than 1000 mg/kg.
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CASE REPORT Top

Pulmonary tuberculosis and Warthin's tumor: Is this a sign for the underlying malignancy? p. 61
Swetabh Purohit, Varsha Joshee, Nishant Chauhan
DOI:10.4103/bbrj.bbrj_143_18  
The enigmatic relationship between pulmonary tuberculosis (TB) and lung cancer has long been attracted clinicians and researchers alike. Knowing the dismal prognosis of advanced bronchogenic carcinoma, efforts are always made for its early diagnosis. Here, we report an interesting case of a 57-year-old patient in which three different disease entities, namely pulmonary TB, lung cancer, and salivary gland tumor, were present concurrently. Premature diagnostic closure led to delay in the diagnosis of bronchogenic carcinoma.
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LETTER TO EDITOR Top

A rare cause of massive hemoptysis in pulmonary tuberculosis – Rasmussen's aneurysm: A forgotten scourge p. 65
Swetabh Purohit, Varsha Joshee
DOI:10.4103/bbrj.bbrj_6_19  
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