bluuebunny/crossref_metadata_2025 huggingface dataset:
Cationic peptidomimetics derived from CAP-18 cathelicidin shown activity in vitro against mycobacterium tuberculosis H37RA and synergy combined with isoniazid
Authors: Chingaté S , Salazar L , Guzmán Fanny
Published: January 2021

Aims & objectives:

Tuberculosis (TB) is the infectious disease that results in the largest number of deaths around the world caused by a bacterial infectious agent. According to the World Health Organization (WHO), there were 10 million new cases and 1.2 million deaths caused by TB in 2018. The problem increases due to drug resistance and HIV/AIDS coinfection, which is why it is necessary to develop new anti-TB compounds with different mechanisms of action. Antimicrobial peptides are molecules gener42ated in the innate immune response of the host with an effect on pathogens. This is the case of the cathelicidins present in mammals involved in the formation of channels in the bacterial cell membrane that leads to cell lysis.

Methods:

In this study, the antimycobacterial activity and the possible mechanism of action of peptides derived from CAP-18 with unnatural amino acids were evaluated. Using bioinformatic tools, the epitope of the native sequence with the best characteristics of helical structure, amphipathicity and antibacterial value was determined. The corresponding peptides, CAP18-B and CAP18-C, had changes from L-Lys to D-Lys and from L-Gly to N-methyl-Gly in the sequence, generating the peptides CAP18-BD, CAP18-CD and the peptoid CAP18-BP. Peptides and peptoids were synthesized using the Fmoc strategy, purified by RP-HPLC and characterized by ESI-MS, circular dichroism and H1 -NMR. The in vitro assesament of the designed sequences was carried out in Mycobacterium smegmatis mc2 155 and

Results:

Mycobacterium tuberculosis H37Ra, obtaining a minimum inhibitory concentration increase from 2 to 6 times in the ranges from 1200 to 60 μg/mL, less than 10% hemolytic activity and no cytotoxic activity on U937 monocytes with LC50 values at least 10 times higher than the MIC and synergy combined with isoniazid. Confocal microscopy showed the insertion of peptides and peptoids in Mycobacterium smegmatis mc2 155 and transmission electron microscopy showed alterations in the cell membrane of Mycobacterium smegmatis mc2 155 evidenced in the formation of blisters, microspheres, channels and cell lysis.

Conclusions:

The results suggest that analogous sequences derived from CAP-18 can be considered as potential antimycobacterial compounds.
Transmissibility study of two vaccine candidates against bovine tuberculosis
Authors: Muñiz Ximena Ferrara , García Elizabeth , Blanco Federica Carlos , Bigi Fabiana , Zumarraga Martín José , Cataldi Angel , Eirin María Emilia
Published: January 2021

Aims & Objective:

We studied the transmissibility in mice of two candidate vaccine M. bovis strains, M. bovis Δmce2 and M. bovis Δmce2ΔphoP, attenuated by gene knock out. The parental strain M. bovis UK (NCTC10772) and a M. bovis wild boar isolate (Mb04-303) were inoculated as controls.

Methods:

Groups of animals including 3 non-inoculated + 3 subcutaneously inoculated with every strain (1.105 CFU) were placed in the same cage (in duplicate). Necropsy was performed at 60 and 90 days post inoculation (dpi) in search of visible tuberculosis lesions (VTL) in lungs and spleens. Organs were analyzed to determine the bacterial load.

Results:

None of the animals presented VTL and no M. bovis was detectable in the non-inoculated group at 60 and 90 dpi. M. bovis was isolated from spleen of all the inoculated mice and from lungs of mice inoculated with control strains. The candidate vaccines were not isolated from lungs.

Conclusions:

This study shows the absence of transmissibility up to 90 days. Recovery of viable control strains from lungs and spleen suggests that the administration route would not alter the capacity of virulent strains to colonize organs. Contrasting, the candidate vaccines were only isolated from spleen. Longer periods should be assayed to evaluate transmissibility of these strains in mice.
IS900 RFLP Analysis of Mycobacterium avium subsp. Paratuberculosis of iranian isolates and analyze using bioinformatics tools
Authors: Khiav Lida Abdolmohammadi , Mosavari Nader , Haghkhah Masoud , Tadayon Keyvan
Published: January 2021

Aims and objectives:

Mycobacterium avium subspecies paratuberculosis was first diagnosed and reported in imported cattle from England during 1962-1965. Further studies have shown that paratuberculosis is scattered throughout Iranian ruminants population. In spite of heavy economic loses; little genetic information is available. So, present study was carried out in order to obtain a better understanding of the MAP genetic diversity in Iranian isolates.

Methods:

During a 7-month period, 36 isolates of MAP was collected from milk and feces obtained from sheep, goat and cattle originating 10 provinces plus a further eleven archived MAP isolates and two MAP vaccine strains (MAP 316F & III &V) and two D4 reference strains (Weybridge and Turkish) were included and digested by restriction endonucleases PstI. Digested DNA was hybridised with a standard IS900 probe. Genomic fingerprints were scanned by camera and analysed using the software Gel pro analyzer & Bionumerix software package version 6.5 (Applied Maths Belgium). Dendrogram and Minimum Spanning Tree was draw using arithmetic averages (UPGMA) for Iranian isolates and vaccine strains.

Results:

ten RFLP (Pst I) types were detected and designated as B (bovine, ovine and caprine), and G (bovine, and caprine) and eight new type (three of them singletons and five cluster) in accordance with the study by pavlik et al 1999. The largest cluster (G strain) contained 45.8% of Iranian isolates (n = 22) then type II (B strain), represented by 11 isolates, recovered from all the three animals in six provinces. A single type was identified only in cattle from three provinces. The identical RFLP type was shown by MAP vaccine strains were different from the Iranian isolates. Two D4 reference strains had restriction pattern but did not hybridize to IS900 sequence. Dendrogram result showed B profiles close to G profile and MAP vaccine strains profiles (>98). The results of Minimum Spanning Tree also showed that all of the clusters and orphan were derived directly or indirectly from B profile with multiple hosts throughout the country. This pattern has been seen from throughout the world and multiple hosts.

Conclusions:

RFLP analysis showed that the Iranian isolates have perhaps originated from B specific profile but it still need to more future works using genotyping methods.
Performance comparison of GenoTypeMTBDRs/Version 1 and version 2 for rapid detection of resistance to fluoroquinolones and injectable drugs among multidrug-resistant mycobacterium tuberculosis in a low MDR-TB setting
Authors: Mokaddas Eiman , Mishari Noura , Ahmed Suhail
Published: January 2021

Aims and objectives:

Fluoroquinolones (FQs) are potent antibiotics against multidrug-resistant Mycobacterium tuberculosis (MDR-TB) and resistance to FQs is associated with higher patient mortality. Rapid and accurate drug susceptibility testing (DST) against FQs and injectable drugs is essential for successful treatment of MDR-TB. FQ resistance mutation detection is equivalent to culture-based DST for predicting MDR-TB treatment outcome. This study compared performance of GenoTypeMTBDRsl Version 1 and Version 2 for detection of resistance to FQs and injectable drugs among MDR-TB strains in Kuwait, a country with low incidence of MDR-TB.

Methods:

MDR-TB strains (n=93) cultured from 74 pulmonary and 19 extra-pulmonary specimens from 93 patients were analyzed by GenoTypeMTBDRsl Version 1 and Version 2 for FQ and injectable drug resistance-associated mutation detection. Repeat isolates from 22 patients, 50 pansusceptible isolates and M. tuberculosis H37 Rv (as control) were also tested. Results were confirmed by PCR-sequencing of gyrA, gyrB, rrs, embB and eis genes. Fingerprinting of isolates was done by spoligotyping.

Results:

Among 93 MDR-TB strains, GenoTypeMTBDRsl Version 1 with/without PCR-sequencing identified 10 isolates with gyrA mutation (A90V, n=2; D94G, n=4; D94A, n=2; D94N, n=1; D94Y, n=1), seven isolates with rrs mutation (A1401G) and 51 isolates with an embB mutation. GenoTypeMTBDRsl Version 2 with/without PCR-sequencing confirmed Version 1 results for gyrA/rrs genes and additionally identified two isolates with gyrB mutation (T500N, n=1; N538D, n=1) and one isolate with eis mutation (-10 G/A). Repeat isolates yielded expected results. No mutation was detected in 50 pansusceptible isolates. Spoligotyping identified 10 Shared International Types among 12 isolates with gyrA/B mutation.

Conclusions:

Our data show that GenoTypeMTBDRsl Version 2 is superior to Version 1 as it identified two additional isolates with gyrB mutations. Improved detection of FQ resistance by GenoTypeMTBDRsl Version 2 will help in proper management of MDR-TB patients in low MDR-TB settings. Supported by KURS grant MI01/18.
Forecasting the Incidence of Tuberculosis in Algeria
Authors: Selmane Scherhazad
Published: January 2021

Aims and objectives:

Tuberculosis (TB) remains a health public issue in Algeria. In spite of the great efforts made for controlling the disease, the number of new reported cases exceeds the twenty thousand cases per year. One worthy of note epidemiological characteristics of TB is its seasonality. The seasonality of TB has been explored in some countries but no study has been conducted in Algeria. The aim of this study is to find out the seasonal pattern and the trend of TB incidence in Algeria.

Methods:

The monthly TB notification data from January 2008 to December 2017 extracted from the National Health Tuberculosis Register were examined to find out the seasonal pattern and trend of TB in Algeria. The seasonal autoregressive integrated moving average (SARIMA) model was used in analysing and predicting TB cases from 2008 to 2016. Cases registered in 2017 were used to assess the prediction accuracy of the selected models. The metrics mean square error (MSE) and mean absolute error (MAE) were applied to assess the better performance of prediction between the selected models.

Results:

From January 2008 to December 2017 there were 215,581 TB notified cases in Algeria with a yearly average 21,467 cases leading to an average annualized morbidity rate of 56,7 per 100,000 inhabitants. The monthly mean was 1796.5, the maximum number (2499) of reported cases was registered in April 2015 and the minimum (1202) in November 2009. The monthly TB data are normally distributed. The incidence of notified pulmonary TB cases decreased from 24 cases per 100,000 inhabitants in 2008 to 15 cases per 100,000 inhabitants in 2017; thus registering a drop of 37.6%, whereas the incidence of notified extrapulmonary TB cases increased from 27.5 cases per 100,000 inhabitants in 2008 to 35.8 cases per 100,000 inhabitants in 2017; thus registering an increase of 29.9%. The heatmap of TB incidence per quarter per province showed a significant temporal and geospatial variation. Time series analysis revealed seasonality with peaks in spring and summer and troughs in autumn and winter and showed that a (3,1,6)×(1,0,1)12 SARIMA model offered the best fit to the TB surveillance data for the period 2008-2016. This model was used to predict TB cases for the year 2017, and the fitted data showed considerable agreement with the actual data.

Conclusions:

Our findings are optimistic for forecasting TB by means of SARIMA models and afford useful information to public health policy makers in formulating targeted prevention and preparedness measures.
Migrating mycobacteriology laboratories
Authors: Shakoor Sadia , Ahmed Imran , Hasan Rumina
Published: January 2021

Aims and objectives: Commissioning and transferring services to new facilities can be a challenge and an opportunity for enhancing services for mycobacteriology laboratories. Challenges are especially prohibitive in low resource settings; however these create an opportunity to develop low cost solutions for maintaining quality reporting while staggering facility based services for laboratories. Since mycobacterial cultures require longer incubation times, commissioning of new facilities and decommissioning older facilities requires careful and time sensitive planning so that services run uninterrupted.
Identification of drug resistant tuberculosis
Authors: Varma-Basil Mandira
Published: January 2021

Aims and objectives:

Multidrug resistant TB (MDR-TB) and extensively drug resistant (XDR) are responsible for significant obstruction to global control of tuberculosis (TB). Drug resistance in Mycobacterium tuberculosis mainly occurs through chromosomal mutations in genes encoding for drug target or drug activating enzymes. However, alternative mechanisms, such as efflux pumps may also be responsible for drug resistance.

Methods:

Insight into the mechanisms of drug resistance would help in limiting the disease through the use and development of better diagnostic and therapeutic tools. The importance of rapid and effective methods of identification of drug resistance in M. tuberculosis cannot be over emphasized. Traditional laboratory techniques are time consuming and cumbersome and not sufficiently sensitive or specific to fight this menace.

Results:

Though rapid molecular methods such as GeneXpert and line probe assays are vital tools in the fight against drug resistant TB, major advances in next generation sequencing (NGS) technology are allowing increasingly rapid and accurate sequencing of entire bacterial genomes, providing unprecedented depth of information.

Conclusion:

NGS has the ability to cause a revolutionary paradigm shift in the diagnosis of drug resistant tuberculosis and in understanding the evolution of drug resistance. However, certain challenges need to be overcome to enable the use of this promising technology in routine diagnosis and research, especially in resource limited regions of the world.
Clinical characteristics and outcome of tuberculous meningitis in the pediatric population- a cross sectional study at the aga khan university hospital
Authors: Qamar Farah , Ahmad Naveed , Mir Fatima
Published: January 2021

Aims and Objectives:

The aim of this retrospective chart review was to determine the clinical characteristics and treatment outcome of the patients diagnosed with Tuberculous Meningitis (TBM) at a tertiary care hospital setting.

Methods:

This was a cross sectional study included children of either gender aged 1-18 years treated for suspected or confirmed diagnosis of TBM at the pediatric unit of Aga Khan University Hospital. The patients enrolled from January 2015 to June 2020 were identified through electronic medical record system. The data was collected on a predesigned paper-based questionnaire and results are presented as mean, standard deviation and interquartile ranges for continuous variables. Whereas, the qualitative variables presented as frequency and percentages.

Results:

There were 52 children included in the study with a mean age of 10.66 ± 6.21 years. Of these 27 (52%) were males and 25 (48%) females. Most of the patients 33 (63.5%) presented within ≤14 days of the onset of symptoms. Median length of stay at hospital was 10 days. The median time from admission to initiation of anti-tuberculous treatment (ATT) was 5 days however, the ATT was initiated before admission to hospital in 11 (21%). In 19 (35%) of patients. Cerebrospinal fluid (CSF) culture was positive for TBM in 5 (9%) patients. GeneXpert test performed on CSF sample in 40 (77%) children. Treatment was completed in half of the patients, seven each lost to follow up and died during treatment. A significant number of patients 33 (63%) developed mild to moderate disability in the initial phase followed by full recovery.

Conclusion:

The findings revealed that TBM is highly associated with morbidity and disability. Early initiation of treatment and diagnosis likely to play pivot role in outcomes.
Adolescent tuberculosis
Authors: Kranzer Katharine
Published: January 2021

Aim and objectives:

Adolescence is characterised by a dramatic increase in the risk of developing tuberculosis, a fact that has been appreciated since the early 20th century. The majority of the world's adolescents live in low and middle-income countries where tuberculosis remains common. It is increasingly apparent that adolescents lie at the heart of the global tuberculosis epidemic, but they have not yet been addressed as a distinct population in tuberculosis policy or within tuberculosis treatment services.

Methods:

We will review the epidemiology of Mycobacterum tuberculosis (M.tb) infection among 10-24 year olds globally and in high tuberculosis burden countries.

Results:

We will discuss the management of M.tb infection and disease, including HIV co-infection and multidrug resistant tuberculosis.

Conclusions:

In addition, we will highlight special considerations regarding the tuberculosis continuum of care during adolescents.
Host-directed Therapy Alleviates Intracellular Mycobacterial Infection via Mediating Innate Immune Responses
Authors: Hussain Tariq , Zhao Deming , Ali Shah Syed Zahid , Sabir Naveed , Wang Jie , Song Yinjuan , Dong Haodi , Mangi Mazhar Hussain , Ni Jiamin , Lifeng Y , Zhou Xiangmei
Published: January 2021

Aims and objectives:

Mycobacterium bovis (M. bovis) is the causative agent of bovine tuberculosis; however, it also causes infection in a wide range of hosts including human beings. Johne's disease (JD) is characterized by chronic granulomatous enteritis predominantly observed in ruminants, caused by Mycobacterium avium subsp. paratuberculosis (MAP). Autophagy is an innate immune defense mechanism for the control of intracellular bacteria, regulated by diverse signaling pathways. It plays a key role in the innate immune defense mechanism for controlling intracellular mycobacterium. Nilotinib, a tyrosine kinase inhibitor (TKI), has been studied extensively in various tumor models; however, no information exists about the pharmacological action of nilotinib in bacterial infections. We hypothesized to investigate the effect of nilotinib on the regulation of host innate immune responses against Mycobacterial infection. In addition, to devise best possible therapeutic remedy towards achieving the goal of mycobacterial diseases treatment based on molecular mechanism both in vitro and in vivo, which may provide useful strategy to treat animal's tuberculosis.

Methods:

In the current study, we conducted both in vitro and in vivo experiments to investigate the role of nilotinib in the regulation of autophagy. Western blot, confocal microscopy, electron microscopy and immunohistochemistry were performed to investigate the induction of autophagy upon nilotinib treatment during M. bovis infection. Gross and histological analysis were conducted to observe the effect of nilotinib on the development of lung's lesions in mice infected with M. bovis. CFU assay was performed to determine the effect of nilotinib on the intracellular growth of M. bovis.

Results:

We found that nilotinib improved the host immune responses by inhibiting intracellular survival and growth of both M. bovis and MAP. Nilotinib also attenuated PI3k/Akt/mTOR axis for the regulation of autophagic degradation of intracellular mycobacterium mediated by inhibition of Abelson (c-ABL) tyrosine kinase. In addition, nilotinib promoted ubiquitination of intracellular bacteria through activation of E3 ubiquitin ligase parkin. Similarly, lung tissue sections of nilotinib treated mice also showed high intensity of parkin protein post M. bovis infection. From in-vivo experiment, we found that nilotinib effectively controlled M. bovis growth and the severity of disease in infected mice.

Conclusions:

Altogether, our data shows that nilotinib regulates protective host innate immune responses against intracellular pathogen both in-vitro and in-vivo and can be exploited as a novel host-directed therapeutic agent in the control of M. bovis and MAP infections.
Hepatitis C Virus and Helicobacter pylori infections in Patients with Pulmonary Tuberculosis
Authors: Nasir Ammara , Rahim Kashif , Ameen Fuad , Ashraf Waqas , Rehman Asad ur , Shehzad Faisal , Alwakeel Suaad S. , Mansoor Muhammad Khalid , Hussian Firasat
Published: January 2021

Aims and objectives:

The control of pulmonary tuberculosis (PTB) mainly depends upon prompt diagnosis and effective treatment which can be hindered by co-infections affecting organs like gastrointestinal system and liver involved in drug bioavailability and metabolism. Among those, Helicobacter pylori (H. pylori) causes chronic gastritis leading to peptic ulcer and related gastric malignancies and liver malfunctions induced by Hepatitis C virus (HCV) co-infections increase the risk of anti-tuberculosis drug-induced hepatotoxicity. The rates of Hepatitis C virus and Helicobacter pylori infections seem to increase day by day in pulmonary tuberculosis patients which need proper investigation followed by timely control and prevention form disease.

Methods:

The hospital-based study was performed in the area of Southern Punjab, Pakistan. The Tehsil Headquarter hospital in the area, where the study was conducted contained TB Clinic. Inclusion and exclusion criteria of suspected patients of pulmonary tuberculosis included in the study. A total of 132 subjects were screened for PTB, HCV and H. pylori infections for finding the role of co-infection. The positive PTB subjects were screened for HCV and H. pylori infections. Both infections (H. pylori and HCV) were also found among TB patients.

Results:

A total of 52.27% were positive for PTB in which 50.72% and 17.39% cases were found in H. pylori and HCV infections respectively. H. pylori had increased rate of infection among PTB cases when compared with that of non-TB cases. HCV was also found to be associated with PTB when compared with non-TB cases. Non-significant gender and age-based differences were present, among study population. Among all PTB patients, HCV and H. pylori coinfections were also found in 5.8% patients in current study.

Conclusions:

Our study showed a high prevalence of HCV and H. pylori infections among PTB patients, which suggest that PTB patients must be screened for the two infections which might be helpful in the control of further complications during the treatment of PTB.
Identification of Cytoplasmic and Conserved Hypothetical Mycobacterium tuberculosis proteins based on mass spectrometry and iedb analysis resource as potent biomarkers for therapeutic strategies or valuable new TB vaccine
Authors: Tasbiti Alireza Hadizadeh , Yari Shamsi , Khanipour Sharareh , Abdolrahimi Farid , Masoumi Morteza , Nematollahi Ali Noor , Ebrahimzadeh Nayyerh , poorazar Shahin , Ghazanfari Morteza , Davari Mehdi , Sakhaee Fatemeh , Siadat Seyed Davar
Published: January 2021

Aims and objectives:

Tuberculosis (TB) remains a major challenge to public health. The disease burden caused by TB is falling globally, in all WHO regions, and in most countries, but not fast enough to reach the first, 2020, milestones of the End TB Strategy. By 2020, the TB incidence rate needs to be falling at 4–5% per year, and the proportion of people with Tuberculosis who die from the disease needs to fall to 10%. These deaths are in part due to late or missed diagnosis and could be prevented with appropriate treatment. The aim of this study was to identify new relevant cytoplasmic and conserved hypothetical protein biomarkers as TB diagnostic or therapeutic targets.

Methods:

TB Protein contents were prepared by detergent phase separation method. Two dimensional gel electrophoresis (2DE) was performed using the Ettan IPGphor 3 isoelecteric focusing (GE Healthcare). Comprehensive genomic and proteomic data were retrieved from MycoBrowser portal (http://www.mycobrowser.epfl.ch) followed by matrix assisted laser desorption ionization time-of flight Mass Spectrometry. The predictive of the MHC I peptide binding is carried out by the IEDB analysis resource (http://www.iedb.org).

Results:

At least six purified proteins with assigned putative functions were determined that corresponded to aldehyde dehydrogenase (Rv0147), iron regulated H-NS (Rv3597c), translocase SecE2 (Rv0379), methyltransferase (Rv3699), adenosylmethionin (Rv1392) and conserved hypothetical protein of Rv0443. Within the category of cytoplasm, membrane and conserved hypothetical patterns, six identified proteins were shown high expression during exponential growth in middlebrook 7H9 broth media. The MHC I peptide binding predictions of these protein spots on the SDS page gels were valued by the Immune Epitope Database tools. The IEDB-AR prediction result is given based on IC50nM value and percentile grad. Principally, a lower score, specified higher affinity. Peptides with IC50 estimation < 50 nM are regarded as high affinity.

Conclusions:

The output scores of the predicting peptides have been shown strong binding for each of identified peptides. Rank of the predicted output affinity of Rv0443 compared to the two others, Rv0147 and Rv0379, presumably leads to stronger binding affinity. The conserved hypothetical proteins, Rv3699 and Rv0443 displayed significant homology with several other TB proteins. These proteins can also be included in TB vaccine constructs or subunit vaccine mixtures combined with a potent adjuvant. We considered these protein profiles to represent reputed biomarker as a new TB vaccine or therapeutic strategies against TB.
Molecular characterization of Mycobacterium tuberculosis isolates using Spoligotyping method in Kurdistan province, iran
Authors: Ramazanzadeh Rashid , Shakib Pegah , Roohia Samaneh , Mohammadi Bahman , Mohajeri Parviz , Borji Soroush
Published: January 2021

Aims and objectives:

Mycobacterium tuberculosis (M. tuberculosis) is one of the major threatening infectious diseases and public health in the world. Spoligotyping is a method for r genotyping assay and molecular epidemiology of M. tuberculosis strains. The aim of this study was to determine the molecular epidemiology of M. tuberculosis by spoligotype isolated from patients with tuberculosis at Kurdistan province, Iran.

Methods:

a total of 47 M. tuberculosis isolates were collected from TB patient at Kurdistan province, Iran. All sample cultures on Lowenstein-Jensen medium and were incubated at 37 °C and were kept for 8 weeks. Bacterial isolates were identified as M. tuberculosis using standard biochemical tests. Multi-drug resistance (MDR) were determined by using proportional method. Spoligotyping was performed.

Results:

Spoligotyping of 47 strains created 8 patterns. Out of 47 samples, 23 (48.94%) were new Spoligotype International Type (SIT). In preexisting shared pattern STI 2669 were highest degree 23.40% that belong to Ural family of M. tuberculosis spoligotypes. Frequency of MDR were 6.38%. Resistance to Isoniazid and Rifampin were 14.89% and 10.64%, respectively.

Conclusion:

In overall, half of isolated strains from patients showed new patterns that, indicated the dynamic transmission of tuberculosis. Therefore, highlights dominance of differ pattern of tuberculosis distribution in west Iran. The low frequency of resistance is very interested.
Kallikrein 12 regulates innate resistance of murine macrophages against Mycobacterium bovis infection by modulating autophagy and apoptosis
Authors: Sabir Naveed , Hussain Tariq , Zhou Xiangmei
Published: January 2021

Aims and objectives:

Tuberculosis (TB) is a leading infectious cause of mortality worldwide and millions of people continue to develop active TB each year. Mycobacterium bovis (M. bovis) is a member of Mycobacterium tuberculosis (Mtb) complex causing bovine tuberculosis and imposing high zoonotic threat to human health. Kallikreins (KLKs) belong to a subgroup of secreted serine proteases and their role is established in various physiological and pathological processes. Since the KLKs expression has been linked to various cancerous, degenerative and infectious conditions, it is likely that KLKs expression may mediate host immune response against M. bovis infection.

Methods:

In the current study, we investigated the expression of KLK12 in M. bovis infection in vivo and in vitro by using C57BL/6 mice and two types of murine macrophages, RAW264.7 cells and bone marrow derived macrophages (BMDMs). In this study, we used two strains of M. bovis, M. bovis C68004 strain and M. bovis N strain. To define the role of KLK12 in immune response regulation of murine macrophages, we produced KLK12 knockdown BMDMs by using siRNA transfection. We investigated the expression of various proteins through QRT-PCR, Immunoblot and ELISA. Apoptotic cells were detected by flow cytometry. Furthermore, cell viability and phagocytic ability of macrophages was also determined after transfection and infection. CFU assay was performed to know the intracellular survival of M. bovis.

Results:

Our results exhibited an increase in the expression of KLK12 in the lung and spleen of M. bovis infected mice. A dose (MOI) and time dependent up-regulation was also noticed in RAW264.7 and BMDMs. Then, we evaluated the role of KLK12 in innate immune response regulation against M. bovis. Interestingly, knockdown of KLK12 resulted into significant down regulation of autophagy and apoptosis in M. bovis infected BMDMs. Furthermore, we demonstrated that this KLK12 mediated regulation of autophagy and apoptosis involves mTOR/AMPK/TSC2 and BAX/Bcl-2/Cytochrome c/Caspase 3 pathways, respectively. Similarly, inflammatory cytokines IL-1β, IL-6, IL-12 and TNF-α were also significantly down regulated in KLK12 knockdown macrophages but difference in IL-10 and IFN-β expression was non-significant. Our finding also depicted that knockdown of KLK12 enhances the intracellular survival of M. bovis.

Conclusion:

Taken together, these findings suggest that up-regulation of KLK12 in M. bovis infected murine macrophages plays a substantial role in the protective immune response regulation by modulating autophagy, apoptosis and pro-inflammatory pathways. To our knowledge, this is the first report on expression and the role of KLK12 in M. bovis infection and the data may contribute to a new paradigm for diagnosis and treatment of bovine and human TB.
Successful management of multidrug-resistant tuberculosis with shorter treatment regimen in niger
Authors: Piubello Alberto , Souleymane Mahamadou Bassirou , Maman-Lawan Ibrahim , Attaher Sala , Boukary Ibrahim , Morou Soumaila , Hassane-Harouna Souleymane
Published: January 2021

Aims and objectives:

Niger is a challenging environment, with limited resources for TB control. Health staff are scarce, access to care is limited due to long distances between communities and health facilities and security threats.

Methods:

Sputum specimens were collected before treatment initiation and monthly for smear and culture during treatment. Follow-up after cure continued up to one year after cure with smear and culture performed every 6 months.

Results:

Among 1,550 patients tested for rifampicin resistance, mainly previously treated patients, 411 (26.5%) were diagnosed with pulmonary RR-TB. The proportion of retreatment cases tested for RR-TB increased from 11.2% in 2008 to 60.5% in 2017. Of the 411 patients with confirmed RR-TB, 359 (87.3%) were enrolled on treatment and 324 (90.3%) started on STR. 24 patients received a modified STR mainly because of hearing loss at baseline or during treatment. Among 300 patients on standardised STR, 250 (83.3%) (IC95%: 78.7-87.1) were cured relapse-free, ten (3.3%) had failure, 28 (9.3%) died, eight (2.7%) were lost to follow-up and four (1.3%) relapsed. Among the 14 patients with bacteriological unfavourable outcome (failure and relapse), nine had initial resistance to fluoroquinolones (FQ) and five had susceptible strains. Among 251 patients with initial susceptibility to FQ there were 4 (1.6%) acquisition of resistance. Of 24 patients receiving a modified STR, 21 (87.5%) were cured relapse-free. Serious ototoxicity was reported in 6 of 120 patients (5%) with clinical monitoring, audiometry at the end of the intensive phase and switch from kanamycin to capreomycin and in 2 of 74 patients (2.8%) with bimonthly audiometry and reduction of kanamycin dose from daily to thrice a week.

Conclusions:

A comprehensive nationwide approach to multidrug-resistant tuberculosis management using the STR was feasible and successful. Hearing loss was manageable replacing the injectable with linezolid. Our study confirms the effectiveness and safety of the STR.
Mapping Mycobacterium tuberculosis genetic diversity in the gulf of guinea – africa
Authors: Thumamo Pokam Benjamin D. , Yeboah-Manu D. , Lawson L. , Guemdjom P. W. , Teyim P. M. , Okonu R. E. , Asuquo Anne E.
Published: January 2021

Aims and objectives:

Tuberculosis (TB) worldwide re-emergence constitutes a major public health problem with multi-drug resistant (MDR) strains becoming a greater threat. Genotyping and drug susceptibility of Mycobacterium tuberculosis complex (MTBC) are important in order to improve the control of the disease. This study sought to determine drug susceptibility and genetic diversity of MTBC in three West/Central African countries, namely Cameroon, Nigeria and Ghana located in the Gulf of Guinea.

Methods:

A collection of 498 isolates (150 from Cameroon, 202 from Nigeria and 146 from Ghana) obtained from culture using Lowenstein–Jensen medium, Middlebrook 7H9 Broth and/or Xpert MTB/RIF and Line Probe Assay (LPA) were included in the study and some isolates tested for drug susceptibility using the proportion method. Isolates were further subjected to IS6110 typing, region of difference 1, 4, 9, 12, 702, 711, spoligotyping and hsp65 gene sequencing analysis.

Results:

The LAM10_CAM was the most prevalent genetic family [128/202 (63.4%) in Nigeria, 46/123 (37.4%) in Ghana and 54/150 (36%) in Cameroon] and SIT 61 the largest cluster with 111/202 (55%) and 43/150 (27%) isolates in Nigeria and Cameroon respectively. Multidrug resistance (MDR) was observed in 29/202 (14.4%) cases in Nigeria, and HIV positivity [37/202 (18.3%) patients] was associated with rifampicin 9/37 (24.3%) resistance (p=0.012), as well as gender (p=0.009). In Cameroon, 13/147 (8.8 %) were resistant to rifampicin (RIF), with 6 (46.2%) of the 13 RIF resistant isolates associated with UgandaI sub-lineage (p=0.0007). Other sublineages such as Haarlem, Ghana, West African 1 were recorded in the three countries.

Conclusion:

Rifampicin resistance in TB/HIV co-infected patient and its association with a sublineage calls for a reinforced surveillance strategy to curb drug resistance development in the Gulf of Guinea.
Implementation of biomedical and social support for people affected by multidrug resistant tuberculosis in guinea
Authors: Hassane H S , Chérif G , Camara S , Diallo D B , Alberto Piubello , Bangoura A , Gutierrez N
Published: January 2021

Aims and objectives:

In 2016, Guinea had an estimated notification rate of 177 new tuberculosis (TB) cases per 100.000 population, with 360 estimated-number of rifampicin-resistant (RR) TB cases. In 2014, Damien Foundation and the National Tuberculosis Programme (NTP) of Guinea started a biomedical-social-support to people treated by multidrug-resistant TB (MDR-TB) in one-pilot health-facility. The aim of this study is to analysis effectiveness of biomedical-social-support on MDR-TB-care.

Methods:

All MDR-TB-cases treated during 2016 to 2017 were analysed. Treatment-outcomes were compared according to the provision of biomedical-social-support in one pilot-health-facility to two-health-facilities without it. In biomedical-social-support, all biological-tests, ancillary drugs were provided free of charge and a nutritional-kit and transport-refunds were monthly provided during the whole treatment. Treatment regimen included 20-month treatment regimen with Kanamycin (Km), Levofloxacin (Lfx), Cycloserine (Cs), Pyrazinamide (Z) and Prothionamide (Pto) during 6-month in the intensive-phase, followed by 12-18-month of same drugs but Km.

Results:

We included 75 MDR-TB cases, 7(9%) HIV-positive. Mean-age was 26years (IQR 15-49). All cases were pulmonary-TB, from which 10(13%) were new-cases. There were 27 MDR-TB cases with biomedical-social support and 48 without it. Mean delay of treatment-start in days was 20(IQR 9-110) in the pilot health-facility compared to 34(IQR 9-111). Treatment outcomes in the group with biomedical-social support were: cured 22(82%), treatment-completion 0(0%), death 2(7%), failure 1(4%) and 2(7%) lost-to-follow-up compared to those without biomedical-social support 23(48%), 2(4%), 9(19%), 2(4%) and 12(25%) respectively. Treatment success to unfavourable- outcomes (failure, death and lost-to-follow-up) in the pilot health-facility was 82% and 18% respectively compared to 52% and 48% respectively in those health-facilities without biomedical-social support (p<0.01).

Conclusions:

The introduction of biomedical-social support to people affected by MDR-TB was successful in Guinea. People who benefited from this strategy had more favourable treatment-outcomes. The biomedical-social support could improve treatment-success if extended to all MDR-TB people under treatment.
Efficacy and safety of intravenous chemotherapy during intensive treatment phase in patients with newly diagnosed pulmonary tuberculosis
Authors: Butova Tetyanas S , Butov Demytro O.
Published: January 2021

Aims and objectives:

The purpose of our study was to examine the efficacy and safety of intravenous chemotherapy during intensive treatment phase in patients with newly diagnosed pulmonary tuberculosis (pulmonary TB).

Methods:

The study involved 92 patients with newly diagnosed pulmonary TB aged between 20 and 68. All patients with newly diagnosed pulmonary TB and chemosensitive tuberculosis were enrolled in the study. The patients were allocated to two groups. The first (control) group of 46 patients received standard chemotherapy orally. The second (main) group consisted of 46 patients who were prescribed isoniazid, rifampin, ethambutol by i.v. infusion, and pyrazinamide orally as a part of the standard treatment.

Results:

Symptoms of intoxication and chest manifestations in pulmonary TB patients from the second group were eliminated faster than the same symptoms in the group 1. In the group 2, the mycobacterial clearance in sputum smears was achieved more rapidly, and up to 2 months it was reached in 37 patients (80.43%), while in the control group in 25 patients (54.35%),p = 0.0066. Destruction healing and inflitrative change alleviation after 4 months was reached in 38 patients (82.61%) (in control group — 28 (60.87%), (p = 0.0192). No additional negative effects were detected when compared with the control group at any time.

Conclusions:

Thanks to i.v. chemotherapy, clinical manifestations of the in-patients with pulmonary TB were eliminated faster, severe side effects of anti-TB drugs were not noticed, time of bacterial clearance and healing of destruction was shorter, healing frequency of destructions increased and the of residual changes decreased.
Acquisition of Cross-Resistance to Bedaquiline and Clofazimine following Treatment for Tuberculosis in Pakistan
Authors: Ghodousi Arash , Rizvi Alamdar Hussain , Baloch Aurangzaib Quadir , Ghafoor Abdul , Khanzada Faisal Masood , Qadir Mehmood , Borroni Emanuele , Trovato Alberto , Tahseen Sabira , Cirillo Daniela Maria
Published: January 2021

Aims and objectives:

In the present study, we report on the first seven cases of acquired bedaquiline and clofazimine resistance identified in Pakistan. The study was conducted within the framework of a retrospective surveillance project to monitor the acquisition of resistance to bedaquiline implemented by the National TB Reference Laboratory in Pakistan and TB Supranational Reference Laboratory.

Methods:

Seventy three sequential isolates from 31 drug-resistant-tuberculosis patients on bedaquiline-containing regimens were retrospectively tested for bedaquiline resistance by MIC testing in 7H11 medium, and for all isolates showing an increased MIC compared to the baseline isolate, further MIC testing for bedaquiline and clofazimine was conducted using the Bactec MGIT960 (BD, Franklin Lakes, NJ, USA). The H37Rv (ATCC 27294) strain was used as a susceptible control in MIC testing. Bedaquiline dry powder was supplied by Janssen-Pharmaceutica (Beerse, Belgium). WGS was carried out with the Illumina Nextera-XT DNA sample preparation kit to prepare paired-end libraries of 150-bp read length to sequence on an Illumina NextSeq platform. Data analysis and single nucleotide polymorphism (SNP) calling were performed using the MTBseq-Pipeline on low-frequency detection mode. Genes associated with resistance to bedaquiline and/or clofazimine (atpE, Rv0678, pepQ, and Rv1979c) were screened for mutations.

Results:

We studied 73 isolates from patients (n=31; 8 MDR, 16 pre-XDR, and 7 XDR) enrolled in bedaquiline-containing regimens. The sequential isolates tested included two strains from 22 patients, three from 7 patients, and four from 2 patients. All baseline strains included in the study were sensitive to bedaquiline. Seven patients developed an increase in bedaquiline MICs in 7H11 medium (range, 0.125 to >0.5mg·liter−1) during therapy, and six of them became resistant to bedaquiline according to the current critical concentration proposed for the drug. In all seven patients we detected genetic variants in Rv0678 that were not present at baseline isolates. Moreover in five out of seven patients the treatment was ultimately failed.

Conclusions:

We documented cases failing therapy that developed specific mutations in Rv0678 and had increased MICs associated with cross-resistance to clofazimine during treatment. This study underlines the relevance of surveillance programs following the introduction of new drugs.
Effectiveness of intravenous isoniazid and ethambutol administration in patients with tuberculosis meningoencephalitis and HIV-infection
Authors: Butov Dmytro O. , Butova Tetyanas S
Published: January 2021

Aims and objectives:

of our study was to investigate the effectiveness of intravenously (i/v) isoniazid (H) and ethambutol (E) administration in patients with new sputum positive drug-susceptible pulmonary tuberculosis (TB) with tuberculous meningoencephalitis (TM) and human immunodeficiency virus (HIV) co-infection in the intensive phase of treatment.

Methods:

Fifty-four patients with TB / TM and HIV co-infection were enrolled in this study. Group 1 included 23 patients treated with E and H i/v, while rifampicin and pyrazinamide were prescribed orally. Group 2 consisted of 31 patients treated with the first-line anti-TB drugs orally. Concentrations of H and E in blood serum were detected using a chromatographic method.

Results:

A significant improvement in clinical symptoms and X-ray signs in patients treated i/v with H and E was observed compared with group 2. Sputum Mycobacterium tuberculosis positivity was observed during the second month of treatment in 25.0 % of patients from group 1 and 76.1% of patients from the control group (p<0.01). In addition, 9 (39.1%) patients died up to six months when H and E were prescribed i/v compared with 22 (70.9%) in group 2 (p=0.02).

Conclusions:

In TB / TM with HIV, i/v H and E treatment was more effective than oral H and E treatment at 2 months of intensive treatment in sputum conversion as well as in clinical improvement, accompanied by significantly higher mean serum concentrations. In addition, the mortality rate was lower in i/v H and E treatment than oral.
Twenty Year Experience of Thoracic Surgery in a Tertiary Institute for Post TB Sequelae
Authors: Dewan Ravindra
Published: January 2021

Aims & Objective:

Post TB sequelae like hemoptysis and recurrent chest infections are a source of lot of morbidity around the globe. Many of these patients present to thoracic surgeons and surgical intervention is able to offer significant relief in many of these patients.

Methods:

Retrospective analysis was carried out for cases operated upon for post TB seqalae in the period from March 1997 to December 2017 in the Thoracic Surgery Department of the National Institute of Tuberculosis and Respiratory Diseases, New Delhi, India. A total of 1325 cases of post TB sequelae were taken up for surgery Out of these, those operated upon primarily for troublesome hemoptysis were 1154 cases. 171 cases were taken up for surgery because they had for recurrent chest infections. Out of these, 558 had tubercular cavities or destroyed lungs, 213 had aspergilloma and post tubercular bronchiectasis was present in 554 cases. Procedures performed were pneumonectomy in 417 patients, lobectomy in 732, bilobectomy in 98, segmental and wedge resections in 16 and thoracoplasty in 62 cases.

Results:

A total of 60 deaths occurred in this series. There were 18 early and 42 late deaths. 7 patients died of hemorrhage, 4 of respiratory failure, 3 sudden deaths in postoperative period (possible cause-Cardiac arrhythmia or cardiac herniation) and 4 of sepsis. Bronchopleural fistula developed postoperatively in 88 cases, which was managed by tube thoracostomy, followed by thoracoplasty, if required. Other complications like wound infection, atelectasis and pneumonitis were quite manageable.

Conclusion:

Overall quite gratifying results are achieved if surgical intervention is offered in patients suffering from debilitating post TB symptoms in carefully selected and well prepared cases.
Value of serologic assay in diagnosis of COVID-19 among patients with negative molecular assay
Authors: Rezaei Mitra , Shiva Parvaneh Baghaei , Sadr Makan , Babamahmoodi Abdolreza , Marjani Majid
Published: January 2021

Aims and objectives:

Due to the growing burden of coronavirus 2019 and considerable rate of RT-PCR false-negative results, a complementary diagnostic tool is necessary for the detection of active cases.

Methods:

This study was performed in Masih Daneshvari Hospital, Tehran, Iran, among admitted symptomatic cases clinically and radiologically suspected to COVID-19 and their first RT-PCR assay was negative. Results of second RT-PCR assay and serologic study were compared.

Results:

In the period of study 137 symptomatic patients with negative RT-PCR assay on the first sample were recruited. Second RT-PCR assay was positive for 45 (32.8%) of them. IgM and IgG antibodies against SARS-CoV-2 were positive in 83 (60.6%) and 86 (62.8%) of patients respectively. Finally, the serologic assay was diagnostic among 73% of patients and the yield of serology was higher after the first week of illness (p-value <0.001). The combination of RT-PCR and serology increased the diagnostic yield to 83.9%.

Conclusions:

This study showed that the serologic assay in parallel to RT-PCR may be useful as a complementary tool for diagnosis of COVID-19 among admitted cases.