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Song Song,1,* Honghong Xu,1,* Jiawei Cao,2,* Guanghong Wu,2 Haiyan Sun,1 Xiaoqi Dai,1 Xuekui Li,1 Meng Chen,1 Menghan Zhang,1 Yueming Yan,1 Jingfeng Tong,3 Zhongdong Wang1 1Institute of Tuberculosis Control, Qingdao Preventive Medicine Research Institute, Qingdao Municipal Center for Disease Control and Prevention, Qingdao, People’s Republic of China; 2Department of Laboratory Medicine, Qingdao Chest Hospital, Qingdao, Shandong, People’s Republic of China; 3Department of Medical Affairs, Shanghai Conlight Medical Co., Ltd., Shanghai, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhongdong Wang, Institute of Tuberculosis Control, Qingdao Preventive Medicine Research Institute, Qingdao Municipal Center for Disease Control and Prevention, No. 8, Fengxian Road, Jinhu Road Street, Shinan District, Qingdao City, Shandong Province, 266000, People’s Republic of China, Email jkzxtb1@163.comPurpose: Nucleotide-based matrix-assisted laser desorption ionization time-of-flight mass spectrometry (nucleotide MALDI-TOF MS) is an emerging molecular technology used for the diagnosis of tuberculosis (TB) caused by Mycobacterium tuberculosis (MTB)and its drug resistance. This study aimed to compare the ability of nucleotide MALDI-TOF MS to detect rifampicin (RIF) resistance in drug-resistant TB (DR-TB) patients with Xpert MTB/RIF and to analyze the disparate results individually. Additionally, potential factors associated with rifampicin resistance among DR-TB patients in Qingdao were investigated.Patients and Methods: A retrospective study was conducted at Qingdao Chest Hospital, and patients with DR-TB were enrolled. Corresponding frozen isolates were recovered and subjected to nucleotide MALDI-TOF MS, Xpert MTB/RIF, and phenotypic drug susceptibility testing (pDST). Sanger sequencing was performed for the discordant results of nucleotide MALDI-TOF MS and Xpert MTB/RIF. Univariate and multivariate logistic regression analyses were used to identify potential factors associated with rifampicin resistance among patients with DR-TB.Results: A total of 125 patients with DR-TB (18.8%, 125/668) were enrolled in this study from May 1 to July 31, 2023. Rifampicin-resistant (DR-TB/RR, 29) and rifampicin-sensitive (DR-TB/RS, 96) groups were divided according to the pDST results. Nucleotide MALDI-TOF MS performed better than Xpert MTB/RIF in terms of sensitivity, specificity, accuracy, and agreement with pDST. Only six cases had inconsistent results, and the sequencing results of five cases were identical to nucleotide MALDI-TOF MS. Furthermore, chest pain (aOR=12.84, 95% CI, 2.29– 91.97, p=0.005), isoniazid sensitivity (aOR=0.14, 0.02– 0.59, p=0.013), and ethambutol sensitivity (aOR=0.02, 0.00– 0.10, p=0.000) were potential factors associated with rifampicin resistance among DR-TB patients in Qingdao.Conclusion: The overall concordance between nucleotide MALDI-TOF MS and Xpert MTB/RIF was 95.2%, with the former performing better in determining rifampicin susceptibility among DR-TB cases in Qingdao. Chest pain, isoniazid, and ethambutol resistance might be factors associated with RIF resistance among patients with DR-TB in Qingdao.Keywords: MTB, rifampicin, nucleotide MALDI-TOF MS, Xpert MTB/RIF, comparison, risk factors

Abstract The study aims to accurately identify differentially expressed genes (DEGs) and biological pathways in mycobacterial infections through bioinformatics for deeper disease understanding. Differentially expressed genes (DEGs) was explored by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Unique DEGs were submitted on least absolute shrinkage and selection operator (LASSO) regression analysis. 1,057 DEGs from two GSE datasets were identified, which were closely connected with NTM/ latent TB infection (LTBI)/active TB disease (ATB). It was demonstrated that these DEGs are mainly associated with detoxification processes, and virus and bacterial infections. Moreover, the METTL7B gene was the most informative marker for distinguishing LTBI and ATB with an area under the curve (AUC) of 0.983 (95%CI: 0.964 to 1). The significantly upregulated HBA1/2 genes were the most informative marker for distinguishing between individuals of IGRA-HC/NTM and LTBI (P

Background: Tuberculosis (TB) remains the number one cause of adult deaths by a curable infectious disease. Mycobacterium tuberculosis bacilli (MTB) is the most common causative organism isolated from mycobacterial lymphadenitis. Accurate and speedy diagnosis is required, especially in paucibacillary extrapulmonary TB. In this study, we compared the efficacy of CBNAAT with line probe assay and liquid culture in the evaluation of tubercular lymph node abscesses. Materials and Methods: This is a hospital-based observational cross-sectional study conducted at Gandhi Medical College and Hospital, Secunderabad, TS, in patients who presented with lymph node abscesses over a period of 18 months after obtaining clearance from the ethics committee and obtaining informed consent. Each sample was evaluated and analyzed in association with the clinical findings, AFB staining, CBNAAT, LPA, and AFB culture, and the results were compared. Results: Fifty patients with TB lymph node abscess diagnosed through FNAC were included in the present study. AFB smear was positive in 29 samples, CBNAAT was positive in 43 samples, LPA was positive in 44 samples, and liquid culture was positive in 39 samples. CBNAAT could detect TB in 16 smear-negative cases, 10 liquid culture-negative cases, and one LPA-negative case. In our study, CBNAAT was more effective in diagnosing TB lymph node abscess than AFB smear. No significant results were obtained when comparing CBNAAT with LPA and liquid culture. Conclusion: This study demonstrates excellent diagnostic accuracy of the Xpert MTB/RIF test in patients with tuberculous lymphadenitis, with a sensitivity of 86%, specificity of 83.33%, negative predictive value of 95.56%, and positive predictive value of 58.82%. Thus, it can be a rule in testing for lymph node TB. These efforts will contribute to the attainment of the TB elimination goal.

Abstract Mycobacterium tuberculosis, the causative agent of tuberculosis, is a pathogenic bacterium that has claimed millions of lives since the Middle Ages. According to the World Health Organization’s report, tuberculosis ranks among the ten deadliest diseases worldwide. The presence of an extensive array of genes and diverse proteins within the cellular structure of this bacterium has provided us with a potent tool for diagnosis. While the culture method remains the gold standard for tuberculosis diagnosis, it is possible that molecular diagnostic methods, emphasis on the identification of mutation genes (e.g., rpoB and gyrA) and single nucleotide polymorphisms, could offer a safe and reliable alternative. Over the past few decades, as our understanding of molecular genetics has expanded, methods have been developed based on gene expansion and detection. These methods typically commence with DNA amplification through nucleic acid targeted techniques such as polymerase chain reaction. Various molecular compounds and diverse approaches have been employed in molecular assays. In this review, we endeavor to provide an overview of molecular assays for the diagnosis of tuberculosis with their properties (utilization, challenges, and functions). The ultimate goal is to explore the potential of replacing traditional bacterial methods with these advanced molecular diagnostic techniques.

Abstract Purpose To assess the eligibility of patients with advanced or recurrent solid malignancies presented to a molecular tumor board (MTB) at a large precision oncology center for inclusion in trials with the endpoints objective response rate (ORR) or duration of response (DOR) based on Response Evaluation Criteria in Solid Tumors (RECIST version 1.1). Methods Prospective patients with available imaging at the time of presentation in the MTB were included. Imaging data was reviewed for objectifiable measurable disease (MD) according to RECIST v1.1. Additionally, we evaluated the patients with MD for representativeness of the identified measurable lesion(s) in relation to the overall tumor burden. Results 262 patients with different solid malignancies were included. 177 patients (68%) had MD and 85 (32%) had non-measurable disease (NMD) at the time point of MTB presentation in accordance with RECIST v1.1. MD was not representative of the overall tumor burden in eleven patients (6%). The main reasons for NMD were lesions with longest diameter shorter than 10 mm (22%) and non-measurable peritoneal carcinomatosis (18%). Colorectal cancer and malignant melanoma displayed the highest rates of MD (> 75%). In contrast, gastric cancer, head and neck malignancies, and ovarian carcinoma had the lowest rates of MD (

The study aims to accurately identify differentially expressed genes (DEGs) and biological pathways in mycobacterial infections through bioinformatics for deeper disease understanding. Differentially expressed genes (DEGs) was explored by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Unique DEGs were submitted on least absolute shrinkage and selection operator (LASSO) regression analysis. 1,057 DEGs from two GSE datasets were identified, which were closely connected with NTM/ latent TB infection (LTBI)/active TB disease (ATB). It was demonstrated that these DEGs are mainly associated with detoxification processes, and virus and bacterial infections. Moreover, the METTL7B gene was the most informative marker for distinguishing LTBI and ATB with an area under the curve (AUC) of 0.983 (95%CI: 0.964 to 1). The significantly upregulated HBA1/2 genes were the most informative marker for distinguishing between individuals of IGRA-HC/NTM and LTBI (P < 0.001). Moreover, the upregulated HBD gene was also differ between IGRA-HC/NTM and ATB (P < 0.001). We have identified gene signatures associated with Mycobacterium infection in whole blood, which could be significant for understanding the molecular mechanisms and diagnosis of NTM, LTBI, or ATB. [ABSTRACT FROM AUTHOR]

N6-methyladenosine (m6A), a well-characterized RNA modification, is involved in regulating multiple biological processes; however, genome-wide identification and functional characterization of the m6A modification in legume plants, including soybean (Glycine max (L.) Merr.), remains lacking. In this study, we utilized bioinformatics tools to perform comprehensive analyses of molecular writer candidates associated with the RNA m6A modification in soybean, characterizing their conserved domains, motifs, gene structures, promoters, and spatial expression patterns. Thirteen m6A writer complex genes in soybean were identified, which were assigned to four families: MT-A70, WTAP, VIR, and HAKAI. It also can be identified that multiple cis elements in the promoters of these genes, which were classified into five distinct groups, including elements responsive to light, phytohormone regulation, environmental stress, development, and others, suggesting that these genes may modulate various cellular and physiological processes in plants. Importantly, the enzymatic activities of two identified m6A writers, GmMTA1 and GmMTA2, were confirmed in vitro. Furthermore, we analyzed the expression patterns of the GmMTAs and GmMTBs under different abiotic stresses, revealing their potential involvement in stress tolerance, especially in the response to alkalinity or darkness. Overexpressing GmMTA2 and GmMTB1 in soybean altered the tolerance of the plants to alkalinity and long-term darkness, further confirming their effect on the stress response. Collectively, our findings identified the RNA m6A writer candidates in leguminous plants and highlighted the potential roles of GmMTAs and GmMTBs in the response to abiotic stress in soybean. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis, the causative agent of tuberculosis, is a pathogenic bacterium that has claimed millions of lives since the Middle Ages. According to the World Health Organization's report, tuberculosis ranks among the ten deadliest diseases worldwide. The presence of an extensive array of genes and diverse proteins within the cellular structure of this bacterium has provided us with a potent tool for diagnosis. While the culture method remains the gold standard for tuberculosis diagnosis, it is possible that molecular diagnostic methods, emphasis on the identification of mutation genes (e.g., rpoB and gyrA) and single nucleotide polymorphisms, could offer a safe and reliable alternative. Over the past few decades, as our understanding of molecular genetics has expanded, methods have been developed based on gene expansion and detection. These methods typically commence with DNA amplification through nucleic acid targeted techniques such as polymerase chain reaction. Various molecular compounds and diverse approaches have been employed in molecular assays. In this review, we endeavor to provide an overview of molecular assays for the diagnosis of tuberculosis with their properties (utilization, challenges, and functions). The ultimate goal is to explore the potential of replacing traditional bacterial methods with these advanced molecular diagnostic techniques. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (Mtb) is an intracellular pathogen capable of adapting and surviving within macrophages, utilizing host nutrients for its growth and replication. Cholesterol is the main carbon source during the infection process of Mtb. Cholesterol metabolism in macrophages is tightly associated with cell functions such as phagocytosis of pathogens, antigen presentation, inflammatory responses, and tissue repair. Research has shown that Mtb infection increases the uptake of low-density lipoprotein (LDL) and cholesterol by macrophages, and enhances de novo cholesterol synthesis in macrophages. Excessive cholesterol is converted into cholesterol esters, while the degradation of cholesterol esters in macrophages is inhibited by Mtb. Furthermore, Mtb infection suppresses the expression of ATP-binding cassette (ABC) transporters in macrophages, impeding cholesterol efflux. These alterations result in the massive accumulation of cholesterol in macrophages, promoting the formation of lipid droplets and foam cells, which ultimately facilitates the persistent survival of Mtb and the progression of tuberculosis (TB), including granuloma formation, tissue cavitation, and systemic dissemination. Mtb infection may also promote the conversion of cholesterol into oxidized cholesterol within macrophages, with the oxidized cholesterol exhibiting anti-Mtb activity. Recent drug development has discovered that reducing cholesterol levels in macrophages can inhibit the invasion of Mtb into macrophages and increase the permeability of anti-tuberculosis drugs. The development of drugs targeting cholesterol metabolic pathways in macrophages, as well as the modification of existing drugs, holds promise for the development of more efficient anti-tuberculosis medications. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) remains the number one cause of adult deaths by a curable infectious disease. Mycobacterium tuberculosis bacilli (MTB) is the most common causative organism isolated from mycobacterial lymphadenitis. Accurate and speedy diagnosis is required, especially in paucibacillary extrapulmonary TB. In this study, we compared the efficacy of CBNAAT with line probe assay and liquid culture in the evaluation of tubercular lymph node abscesses. Materials and Methods: This is a hospital‑based observational cross‑sectional study conducted at Gandhi Medical College and Hospital, Secunderabad, TS, in patients who presented with lymph node abscesses over a period of 18 months after obtaining clearance from the ethics committee and obtaining informed consent. Each sample was evaluated and analyzed in association with the clinical findings, AFB staining, CBNAAT, LPA, and AFB culture, and the results were compared. Results: Fifty patients with TB lymph node abscess diagnosed through FNAC were included in the present study. AFB smear was positive in 29 samples, CBNAAT was positive in 43 samples, LPA was positive in 44 samples, and liquid culture was positive in 39 samples. CBNAAT could detect TB in 16 smear‑negative cases, 10 liquid culture‑negative cases, and one LPA‑negative case. In our study, CBNAAT was more effective in diagnosing TB lymph node abscess than AFB smear. No significant results were obtained when comparing CBNAAT with LPA and liquid culture. Conclusion: This study demonstrates excellent diagnostic accuracy of the Xpert MTB/RIF test in patients with tuberculous lymphadenitis, with a sensitivity of 86%, specificity of 83.33%, negative predictive value of 95.56%, and positive predictive value of 58.82%. Thus, it can be a rule in testing for lymph node TB. These efforts will contribute to the attainment of the TB elimination goal. [ABSTRACT FROM AUTHOR]

MTB trail building and maintenance in Denmark frequently take place as co-production involving volunteers and public authorities. The aim of this paper is to explore how organizational factors of public authorities impact the level and quality of co-production in relation to voluntary MTB trail building. The aim is pursued through content analysis of qualitative interviews with 30 volunteers and representatives for public authorities in three Danish municipalities. The findings show that favourable conditions for voluntary MTB trail building depend on (i) the availability of the public officials; (ii) whether financial, practical and administrative resources are available; (iii) how the public officials perceive the volunteers; (iv) whether possibilities of joint decision-taking between volunteers and public officials exist and (v) to which extent the volunteers have a voice in the decision process and, if so, when they were involved. Finally, the importance of early involvement of volunteers in co-production processes and how the five points can contribute to this are discussed. [ABSTRACT FROM AUTHOR]

This study aimed to compare the degree of epigenetic modifications between a TB-HIV co-infected cohort and uninfected subjects. Formalin-fixed paraffin-embedded (FFPE) tissues were retrieved from 45 TB-HIV co-infected and 45 control individuals. Real-time PCR was applied to compare the level of expression of genes involved in epigenetic regulation. The protein multiplex assay was used to assess the degree of protein modification. DNA sequencing was used to determine the evolutionary relationships between the infecting HIV and Mtb strains. Our results indicated a significant increase in the expression of the five candidate genes in the patients with TB-HIV relative to the control cohort. A sharp increase in the degree of histone methylation, acetylation and phosphorylation was observed in TB-HIV co-infected patients. The phylogenetic analysis classified the strains into three distinct HIV clusters and five Mtb clusters. The disparities in the expression profiles of our candidate genes between the TB-HIV cohort and non-TB-HIV group highlights the important role played by various TB and HIV strains in regulating the host gene expression landscape. [ABSTRACT FROM AUTHOR]

Xue-Song Xiong,1,2,&ast; Xue-Di Zhang,3,&ast; Jia-Wei Yan,3 Ting-Ting Huang,1,2 Zhan-Zhong Liu,4 Zheng-Kang Li,5 Liang Wang,5 Fen Li1,2 1Department of Laboratory Medicine, The Affiliated Huai’an Hospital of Yangzhou University, Huai’an, Jiangsu Province, People’s Republic of China; 2Department of Laboratory Medicine, The Fifth People’s Hospital of Huai’an, Huai’an, Jiangsu Province, People’s Republic of China; 3Department of Laboratory Medicine, Xuzhou Infectious Diseases Hospital, Xuzhou, Jiangsu Province, People’s Republic of China; 4Department of Pharmacy, Xuzhou Infectious Diseases Hospital, Xuzhou, Jiangsu Province, People’s Republic of China; 5Department of Laboratory Medicine, Guangdong Provincial People’s Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou, Guangdong Province, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Liang Wang; Fen Li, Email healthscience@foxmail.com; 3063892835@qq.comAbstract: Multidrug-resistant tuberculosis (MDR-TB) is an essential cause of tuberculosis treatment failure and death of tuberculosis patients. The rapid and reliable profiling of Mycobacterium tuberculosis (MTB) drug resistance in the early stage is a critical research area for public health. Then, most traditional approaches for detecting MTB are time-consuming and costly, leading to the inappropriate therapeutic schedule resting on the ambiguous information of MTB drug resistance, increasing patient economic burden, morbidity, and mortality. Therefore, novel diagnosis methods are frequently required to meet the emerging challenges of MTB drug resistance distinguish. Considering the difficulty in treating MDR-TB, it is urgently required for the development of rapid and accurate methods in the identification of drug resistance profiles of MTB in clinical diagnosis. This review discussed recent advances in MTB drug resistance detection, focusing on developing emerging approaches and their applications in tangled clinical situations. In particular, a brief overview of antibiotic resistance to MTB was present, referred to as intrinsic bacterial resistance, consisting of cell wall barriers and efflux pumping action and acquired resistance caused by genetic mutations. Then, different drug susceptibility test (DST) methods were described, including phenotype DST, genotype DST and novel DST methods. The phenotype DST includes nitrate reductase assay, RocheTM solid ratio method, and liquid culture method and genotype DST includes fluorescent PCR, GeneXpert, PCR reverse dot hybridization, ddPCR, next-generation sequencing and gene chips. Then, novel DST methods were described, including metabolism testing, cell-free DNA probe, CRISPR assay, and spectral analysis technique. The limitations, challenges, and perspectives of different techniques for drug resistance are also discussed. These methods significantly improve the detection sensitivity and accuracy of multidrug-resistant tuberculosis (MRT) and can effectively curb the incidence of drug-resistant tuberculosis and accelerate the process of tuberculosis eradication. Keywords: MTB, antibiotic resistance, Raman spectroscopy, rapid detection

Background: Tuberculosis (TB) is one of the leading infectious diseases that cause death worldwide; the emergence of drug resistant mycobacterium tuberculosis is of great concern for TB control programme in Nigeria. Aim and objective: To detect multi-drug and extensively drug resistant mycobacterium tuberculosis among tuberculosis patients in Kaduna State North west Nigeria. Study design: This was a cross sectional study among tuberculosis patients attending directly observed treatment short course (DOTS) clinics in the three (3) senatorial districts of Kaduna State, Nigeria. Materials and methods: A total of 360 presumptive drug resistant tuberculosis patients’ samples were collected and decontaminated using modified Petroff method (NaOH-Na-citrate-NALC) from August 2018 to December 2020. Drug susceptibility testing was performed on first- and second-line anti-tuberculosis drugs for the 305 confirmed MTBC isolates using Lowenstein Jensen (LJ) proportion method. Results: A total of 67(21.8%) isolates were pan-susceptible, 176(57.5%) isolates were MDR-TB, rifampicin and isoniazid mono-resistant were 46(15.1%) and 17(5.6%) respectively. For the second line drugs, a total of 271(80.22%) isolates were found to be pan-susceptible and 35(19.88%) isolates were pre-extensively drug resistant TB. Conclusions: These findings established higher prevalence of MDR-TB and pre-extensively resistant TB in Kaduna state.

Background: Pulmonary tuberculosis (PTB) is a common infectious disease caused by mycobacterium tuberculosis (MTB) and the present study aims to explore the associations of genetic variants within tyrosine kinases 2 (TYK2) with PTB incidence. Methods: A population‐based case control study including 168 smear‐positive PTB cases and 251 controls was conducted. Five single nucleotide polymorphisms (SNPs) including rs280520, rs91755, rs2304256, rs12720270, rs280519 located within TYK2 gene were selected and MassARRAY® MALDI‐TOF system was employed for genotyping. SPSS 19.0 was adopted for statistical analysis, non‐conditional logistic regression was conducted. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were computed to estimate their contributions to PTB incidence. Results: In the overall study population, rs91755 TT and rs280519 AA genotypes were found to be associated with reduced PTB risk (OR = 0.34, 95% CI: 0.16–0.72; OR = 0.38, 95% CI: 0.18–0.79, respectively). After stratification for sex, we found that among the male population, rs91755TG/TT, rs12720270AG/GG and rs280519AG/AA genotypes were associated with reduced PTB risk (OR = 0.41, 95% CI: 0.21–0.80; OR = 0.44, 95% CI: 0.21–0.94; OR = 0.42, 95% CI: 0.21–0.82, respectively). After stratification for age, we found that among those aged <60 years, rs91755TT and rs280519AA genotype were associated with reduced PTB risk (OR = 0.29, 95% CI: 0.09–0.90; OR = 0.34, 95% CI: 0.11–1.08, respectively); while rs2304256AC/AA genotype was associated with increased PTB risk (OR = 2.68, 95% CI: 1.05–6.85). Haplotype analysis revealed that AGAAG and ATCGA (Combined with rs280520, rs91755, rs2304256, rs12720270 and rs280519) were associated with increased (OR = 1.54, 95% CI: 1.01–2.37) and decreased PTB risk (OR = 0.70, 95% CI: 0.52–0.94), respectively. Conclusions: The genetic variants located within TYK2 including rs91755, rs12720270 and rs280519 were found to be associated with modified PTB risk and the SNPs had potential to be the biomarkers to predict PTB incidence risk. TYK2 modified TB risk. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis (TB) is one of the leading infectious diseases that cause death worldwide; the emergence of drug resistant Mycobacterium tuberculosis is of great concern for TB control program in Nigeria. Aim and Objective: To detect multi-drug and extensively drug resistant Mycobacterium tuberculosis among Tuberculosis Patients in Kaduna State North west Nigeria. Study Design: This was a cross sectional study among tuberculosis patients attending Directly Observed Treatment Short Course (DOTS) clinics in the three (3) senatorial districts of Kaduna State, Nigeria. Material and Methods: A total of 360 presumptive drug resistant tuberculosis patients' samples were collected and decontaminated using modified Petroff method (NaOH-Na-citrate-NALC) from August 2018 to December 2020. Drug susceptibility testing was performed on first- and second-line anti-tuberculosis drugs for the 305 confirmed MTBC isolates using Lowenstein Jensen (LJ) Proportion Method. Results: A total of 67(21.8%) isolates were pan-susceptible, 176(57.5%) isolates were MDR-TB, Rifampicin and Isoniazid mono-resistant were 46(15.1%) and 17(5.6%) respectively. For the second line drugs, a total of 271(80.22%) isolates were found to be pan-susceptible and 35(19.88%) isolates were pre-extensively drug resistant TB. Conclusions: These findings established higher prevalence of MDR-TB and pre-extensively resistant TB in Kaduna state. [ABSTRACT FROM AUTHOR]

N6-methyladenosine (m6A), a well-characterized RNA modification, is involved in regulating multiple biological processes; however, genome-wide identification and functional characterization of the m6A modification in legume plants, including soybean (Glycine max (L.) Merr.), remains lacking. In this study, we utilized bioinformatics tools to perform comprehensive analyses of molecular writer candidates associated with the RNA m6A modification in soybean, characterizing their conserved domains, motifs, gene structures, promoters, and spatial expression patterns. Thirteen m6A writer complex genes in soybean were identified, which were assigned to four families: MT-A70, WTAP, VIR, and HAKAI. It also can be identified that multiple cis elements in the promoters of these genes, which were classified into five distinct groups, including elements responsive to light, phytohormone regulation, environmental stress, development, and others, suggesting that these genes may modulate various cellular and physiological processes in plants. Importantly, the enzymatic activities of two identified m6A writers, GmMTA1 and GmMTA2, were confirmed in vitro. Furthermore, we analyzed the expression patterns of the GmMTAs and GmMTBs under different abiotic stresses, revealing their potential involvement in stress tolerance, especially in the response to alkalinity or darkness. Overexpressing GmMTA2 and GmMTB1 in soybean altered the tolerance of the plants to alkalinity and long-term darkness, further confirming their effect on the stress response. Collectively, our findings identified the RNA m6A writer candidates in leguminous plants and highlighted the potential roles of GmMTAs and GmMTBs in the response to abiotic stress in soybean.

Background: Co-infection of Mycobacterium tuberculosis (MTB) and the Human Immunodeficiency Virus (HIV) is a major global public health issue, particularly in border areas of resource-limited nations, including Ethiopia. Objective: To explore the prevalence and associated variables of MTB/HIV co-infection among PTB presumptive patients in Northwest Ethiopia. Methods: From February to August 2021, a cross-sectional institutional investigation was conducted at the Metema and Abrehajira hospitals. Semi-structured questionnaires were used to collect socio-demographic and clinical data. The MTB/RIF Xpert assay was used to process sputum, and 3 ml of veins blood was collected for HIV rapid test (STAT-PAK, ABON, and SD BIOLINE HIV test algorithm) following the Ethiopian National HIV test algorithm. The Gene Xpert assay's sample processing control was checked to ensure data quality. Data entered into Epi-Data were exported to SPSS version 20 for analysis. Statistically significant variables (p-value ≤0.2) from bivariable analysis were included in multivariable analysis. A p-value ≤ 0.05 was judged statistically significant. Results: This study included 314 PTB presumptive patients with a median age of 35.0 years, of which 178 (56.69 %) were males. Among all patients, 40(12.7 %) and 51(16.2 %) were PTB, and HIV seropositivity, respectively. Of the PTB patients, 14/40 (35 %) (95 % CI: 24.4–45.6) were co-infected with HIV/AIDS. Married patients were 70 % less likely than unmarried individuals (AOR = 0.3 CI; 0.07–0.98) to have MTB/HIV co-infection. Patients who had contact history with MDR-TB patients (AOR = 5 CI; 1.37–18.00), and those who had a history of alcohol use (AOR = 12.2 CI; 2.56–57.8) were more likely to have MTB-HIV co-infection than their peers. Conclusion: Our findings showed that MTB-HIV co-infection is one of the most important community health concerns in the study area. Therefore, MTB/HIV cooperation activities should be fully in place to prevent co-infection and its impact on the population.

Abstract Background In neuro-oncology, the inclusion of tumor patients in the molecular tumor board has only become increasingly widespread in recent years, but so far there are no standards for indication, procedure, evaluation, therapy recommendations and therapy implementation of neuro-oncological patients. The present work examines the current handling of neuro-oncological patients included in molecular tumor boards in Germany. Methods We created an online based survey with questions covering the handling of neuro-oncologic patient inclusion, annotation of genetic analyses, management of target therapies and the general role of molecular tumor boards in neuro-oncology in Germany. We contacted all members of the Neuro-Oncology working group (NOA) of the German Cancer Society (DKG) by e-mail. Results 38 responses were collected. The majority of those who responded were specialists in neurosurgery or neurology with more than 10 years of professional experience working at a university hospital. Molecular tumor boards (MTB) regularly take place once a week and all treatment disciplines of neuro-oncology patients take part. The inclusions to the MTB are according to distinct tumors and predominantly in case of tumor recurrence. An independently MTB member mostly create the recommendations, which are regularly implemented in the tumor treatment. Recommendations are given for alteration classes 4 and 5. Problems exist mostly within the cost takeover of experimental therapies. The experimental therapies are mostly given in the department of medical oncology. Conclusions Molecular tumor boards for neuro-oncological patients, by now, are not standardized in Germany. Similarities exists for patient inclusion and interpretation of molecular alterations; the time point of inclusion and implementation during the patient treatment differ between the various hospitals. Further studies for standardization and harmonisation are needed. In summary, most of the interviewees envision great opportunities and possibilities for molecular-based neuro-oncological therapy in the future.

Background: In neuro-oncology, the inclusion of tumor patients in the molecular tumor board has only become increasingly widespread in recent years, but so far there are no standards for indication, procedure, evaluation, therapy recommendations and therapy implementation of neuro-oncological patients. The present work examines the current handling of neuro-oncological patients included in molecular tumor boards in Germany. Methods: We created an online based survey with questions covering the handling of neuro-oncologic patient inclusion, annotation of genetic analyses, management of target therapies and the general role of molecular tumor boards in neuro-oncology in Germany. We contacted all members of the Neuro-Oncology working group (NOA) of the German Cancer Society (DKG) by e-mail. Results: 38 responses were collected. The majority of those who responded were specialists in neurosurgery or neurology with more than 10 years of professional experience working at a university hospital. Molecular tumor boards (MTB) regularly take place once a week and all treatment disciplines of neuro-oncology patients take part. The inclusions to the MTB are according to distinct tumors and predominantly in case of tumor recurrence. An independently MTB member mostly create the recommendations, which are regularly implemented in the tumor treatment. Recommendations are given for alteration classes 4 and 5. Problems exist mostly within the cost takeover of experimental therapies. The experimental therapies are mostly given in the department of medical oncology. Conclusions: Molecular tumor boards for neuro-oncological patients, by now, are not standardized in Germany. Similarities exists for patient inclusion and interpretation of molecular alterations; the time point of inclusion and implementation during the patient treatment differ between the various hospitals. Further studies for standardization and harmonisation are needed. In summary, most of the interviewees envision great opportunities and possibilities for molecular-based neuro-oncological therapy in the future. [ABSTRACT FROM AUTHOR]

Aims/objectives: A multidisciplinary tumor board (MTB) is a crucial aspect of comprehensive cancer care, providing evidence-based treatment for patients. The aim of this analysis was to assess the importance of MTB in comprehensive cancer care and its impact on cancer patients' management and treatment compliance. Method: One-year audit of the patients discussed on the tumor board of a private comprehensive cancer care hospital between September 2020 and August 2021. The data were collected before, during, and after each MTB meeting. Result: A study of 800 cases discussed on the tumor board in one year. Approximately 60% of cases were presented by surgical oncology, 21% by medical oncology, and the rest by other departments. The median age of patients was 56 years. This audit revealed that the most common tumor was head and neck cancer (28.5%), followed by gastro-intestinal and breast cancer (55.2%). The most common treatment plan was surgical management (57.6%), followed by systemic therapy (45%), radiotherapy (32.1%), and palliative care (14.5%). Treatment compliance was over 50%, and after one year, 52.87% of patients were still alive. The audit revealed that more than 20% of the data was missing. Conclusion: The difficult management of advanced-stage disease is improved, and treatment compliance is increased, through MTB practice. The demographics of cancer in this region of the country were also displayed by this audit. In the future, a prospective trial with a larger patient population, comprehensive follow-up information, and fewer missing data points should be used to evaluate the true impact of multidisciplinary care. [ABSTRACT FROM AUTHOR]

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Monitoring ctDNA by liquid biopsies seems to represent the perfect match for precision oncology and its cornerstone clinical framework: the molecular tumour board (MTB). Detecting and scrutinising the success of targeted therapies or tracking and, for that matter, addressing the therapy with the evolutive nature of a tumour are some of the main advancements one considers to be important for the MTB. One challenge is correlating the estimated allele frequency of each identified genetic alteration determined by analysing the ctDNA sequencing results and matching these with the range of suitable drugs, which may limit the simultaneous treatment of all tumour variations. This limitation arises because a new biopsy would typically be required to evaluate the response to treatment. As a result, evaluating the success of MTB recommendations relies on traditional staging methods, highlighting an existing diagnostic gap. Thus, optimising liquid biopsy technology could enhance the efficacy of MTB treatment recommendations and ensuing tailored therapies. Herein, we discuss the prospect of ctDNA analyses in the molecular tumour board. [ABSTRACT FROM AUTHOR]

Colorectal cancer (CRC) is currently one of the most common tumor types diagnosed worldwide. In the early stages, the disease responds well to surgical and chemotherapeutic treatment, but in the later stages when therapeutic options are exhausted, comprehensive genomic profiling can guide further treatment decisions. We present the case of a 46-year-old man of Ashkenazi Jewish ancestry who was diagnosed with KRAS-mutated metastatic colorectal cancer. After surgery and progression on standard FOLFOX/FOLFIRI + bevacizumab therapy, as well as on Trifluridine/Tipiracil, comprehensive genomic profiling was performed with the hope of expanding therapeutic options. Following comprehensive tumor molecular profiling via NGS, a discussion of the case was discussed at the local molecular tumor board in order to determine further treatment strategy. An activating variant of KRAS and PIK3CA, FLT3 and SRC amplification and damaging TP53 and APC variants were discarded by MTB as potential targetable biomarkers. The BRCA2 p.S1415fs*4 founder frameshift variant was of interest and the patient was included in the clinical trial investigating the efficacy of a PARP inhibitor talazoparib. Unfortunately, the disease progression was detected within one month of talazoparib treatment and the patient died during the 8th cycle of FOLFIRI + bevacizumab therapy rechallenge. [ABSTRACT FROM AUTHOR]

Copyright of Hladnikia is the property of Botanical Society of Slovenia / Botanicno Drustvo Slovenije and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Background Pulmonary tuberculosis (PTB) is a common infectious disease caused by mycobacterium tuberculosis (MTB) and the present study aims to explore the associations of genetic variants within tyrosine kinases 2 (TYK2) with PTB incidence. Methods A population‐based case control study including 168 smear‐positive PTB cases and 251 controls was conducted. Five single nucleotide polymorphisms (SNPs) including rs280520, rs91755, rs2304256, rs12720270, rs280519 located within TYK2 gene were selected and MassARRAY® MALDI‐TOF system was employed for genotyping. SPSS 19.0 was adopted for statistical analysis, non‐conditional logistic regression was conducted. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were computed to estimate their contributions to PTB incidence. Results In the overall study population, rs91755 TT and rs280519 AA genotypes were found to be associated with reduced PTB risk (OR = 0.34, 95% CI: 0.16–0.72; OR = 0.38, 95% CI: 0.18–0.79, respectively). After stratification for sex, we found that among the male population, rs91755TG/TT, rs12720270AG/GG and rs280519AG/AA genotypes were associated with reduced PTB risk (OR = 0.41, 95% CI: 0.21–0.80; OR = 0.44, 95% CI: 0.21–0.94; OR = 0.42, 95% CI: 0.21–0.82, respectively). After stratification for age, we found that among those aged

This study aimed to compare the degree of epigenetic modifications between a TB-HIV co-infected cohort and uninfected subjects. Formalin-fixed paraffin-embedded (FFPE) tissues were retrieved from 45 TB-HIV co-infected and 45 control individuals. Real-time PCR was applied to compare the level of expression of genes involved in epigenetic regulation. The protein multiplex assay was used to assess the degree of protein modification. DNA sequencing was used to determine the evolutionary relationships between the infecting HIV and Mtb strains. Our results indicated a significant increase in the expression of the five candidate genes in the patients with TB-HIV relative to the control cohort. A sharp increase in the degree of histone methylation, acetylation and phosphorylation was observed in TB-HIV co-infected patients. The phylogenetic analysis classified the strains into three distinct HIV clusters and five Mtb clusters. The disparities in the expression profiles of our candidate genes between the TB-HIV cohort and non-TB-HIV group highlights the important role played by various TB and HIV strains in regulating the host gene expression landscape.

En la actualidad la práctica del ciclismo ha tenido un incremento considerable, así como el uso de bicicletas de montaña (Mountain Bike, MTB, en inglés), de cuadro rígido, utilizadas como medio de transporte y para competencia, debido a su costo asequible. Este tipo de bicicletas, al ser utilizadas para varios propósitos, presentan esfuerzos variados en su cuadro, que conllevan a sobrepasar las exigencias de diseño, presentándose fallos en las vainas superiores. Este tipo de fallo es analizado en este estudio, motivo por el cual se levanta la información referente al material del cuadro, cargas actuantes y modelado 3D. Posterior se genera un análisis de homologación del fallo y se determina una propuesta de mejora aplicando optimización geométrica, donde se determina un espesor de 3,50 mm en las vainas superiores, garantizando la resistencia del cuadro de bicicleta bajo las condiciones de estudio; es decir, un drop de 60 cm y carga de 74 kg, con la cual se garantiza que el esfuerzo en las vainas superiores no sobrepase el esfuerzo último del material de 890,94 MPa.

Colorectal cancer (CRC) is currently one of the most common tumor types diagnosed worldwide. In the early stages, the disease responds well to surgical and chemotherapeutic treatment, but in the later stages when therapeutic options are exhausted, comprehensive genomic profiling can guide further treatment decisions. We present the case of a 46-year-old man of Ashkenazi Jewish ancestry who was diagnosed with KRAS-mutated metastatic colorectal cancer. After surgery and progression on standard FOLFOX/FOLFIRI + bevacizumab therapy, as well as on Trifluridine/Tipiracil, comprehensive genomic profiling was performed with the hope of expanding therapeutic options. Following comprehensive tumor molecular profiling via NGS, a discussion of the case was discussed at the local molecular tumor board in order to determine further treatment strategy. An activating variant of KRAS and PIK3CA, FLT3 and SRC amplification and damaging TP53 and APC variants were discarded by MTB as potential targetable biomarkers. The BRCA2 p.S1415fs*4 founder frameshift variant was of interest and the patient was included in the clinical trial investigating the efficacy of a PARP inhibitor talazoparib. Unfortunately, the disease progression was detected within one month of talazoparib treatment and the patient died during the 8th cycle of FOLFIRI + bevacizumab therapy rechallenge.

To date, several emerging biomarkers have gained considerable interest in the field of predictive molecular oncology. The advent of precision medicine has led to the development of innovative drugs targeting rare molecular pathways independently from histology, defined as tissue-agnostic drugs. Although there is a lot of promise for this new tissue-agnostic model in the oncological scenario, crucial issues from both the diagnostic and therapeutic standpoint are emerging. This review aims to critically examine the role of tissue-agnostic biomarkers in different solid tumors, focusing on the prevalence and methods of detection of agnostic biomarkers together with drug approvals to guide clinicians in this evolving landscape. To strengthen the framework for tissue-agnostic approvals, the dialogue between regulatory, industrial, and academic parties should be intensified. Critical questions include the development of an efficient network system that can overcome the heterogeneity of patients' inclusion criteria along with the increasingly difficult interpretation of next-generation sequencing (NGS) profiling technologies. Cost-effectiveness and risk–benefit studies are needed in the national context considering the modalities of access to diagnostic tests and reimbursement of treatments. [ABSTRACT FROM AUTHOR]

Introduction: Neutrophil granulocytes predominate in the lungs of patients infected with Mycobacterium tuberculosis (Mtb) in earlier stages of the disease. During infection, neutrophils release neutrophil extracellular traps (NETs), an antimicrobial mechanism by which a DNA-backbone spiked with antimicrobial components traps the mycobacteria. However, the specific mycobacterial factors driving NET formation remain unclear. Proteins from the proline-glutamic acid (PE)/proline-proline-glutamic acid (PPE) family are critical to Mtb pathophysiology and virulence. Methods: Here, we investigated NET induction by PE18, PPE26, and PE31 in primary human blood-derived neutrophils. Neutrophils were stimulated with the respective proteins for 3h, and NET formation was subsequently assessed using confocal fluorescence microscopy. Intracellular ROS levels and cell necrosis were estimated by flow cytometry. Additionally, the influence of phorbol-12-myristate-13-acetate (PMA), a known NADPH oxidase enhancer, on NET formation was examined. Neutrophil integrity following incubation with the PE/PPE proteins was evaluated using transmission electron microscopy. Results: For the first time, we report that stimulation of primary human blood-derived neutrophils with Mtb proteins PE18, PPE26, and PE31 resulted in the formation of NETs, which correlated with an increase in intracellular ROS levels. Notably, the presence of PMA further amplified this effect. Following incubation with the PE/PPE proteins, neutrophils were found to remain viable and structurally intact, as verified through transmission electron microscopy, indicating the occurrence of vital NET formation. Discussion: These findings offer valuable insights that contribute to a better understanding of host-pathogen interactions during Mtb infection. Moreover, they underscore the significance of these particular Mtb antigens in triggering NET formation, representing a distinctive and previously unrecognized function of PE/PPE antigens. [ABSTRACT FROM AUTHOR]

Antimicrobial peptides (AMPs) can directly kill Gram-positive bacteria, Gram-negative bacteria, mycobacteria, fungi, enveloped viruses, and parasites. At sublethal concentrations, some AMPs and also conventional antibiotics can stimulate bacterial response increasing their resilience, also called the hormetic response. This includes stimulation of growth, mobility, and biofilm production. Here, we describe the discovery of AMPs that stimulate the growth of certain mycobacteria. Peptide 14 showed a growth stimulating effect on Mycobacteria tuberculosis (MTB), M. bovis, M. avium subsp. paratuberculosis (MAP), M. marinum, M. avium-intracellulare, M. celatum, and M. abscessus. The effect was more pronounced at low bacterial inocula. The peptides induce a faster transition from the lag phase to the log phase and keep the bacteria longer in the log phase before entering stationary phase when compared to nontreated controls. In some cases, an increase in the division rate was observed. An initial screen using MAP and a collection of 75 peptides revealed 13 peptides with a hormetic effect. For MTB, a collection of 25 artificial peptides were screened and 13 were found to reduce the time to positivity (TTP) by at least 5%, improving growth. A screen of 43 naturally occurring peptides, 11 fragments of naturally occurring peptides and 5 designed peptides, all taken from the database APD3, identified a further 44 peptides that also lowered TTP by at least 5%. Lasioglossin LL-III (Bee) and Ranacyclin E (Frog) were the most active natural peptides, and the human cathelicidin LL37 fragment GF-17 and a porcine cathelicidin protegrin-1 fragment were the most active fragments of naturally occurring peptides. Peptide 14 showed growth-stimulating activity between 10 ng/mL and 10 µg/mL, whereas the stability-optimised Peptide 14D had a narrow activity range of 0.1–1 µg/mL. Peptides identified in this study are currently in commercial use to improve recovery and culture for the diagnostics of mycobacteria in humans and animals. [ABSTRACT FROM AUTHOR]

Background We evaluated the relationship between response to efflux pump inhibition in fluoroquinolone-resistant Mycobacterium tuberculosis (Mtb) isolates and differences in gene expression and expression quantitative trait loci (eQTL). Methods We determined ofloxacin minimum inhibitory concentration (MIC) for ofloxacin-resistant and -susceptible Mtb isolates without and with the efflux pump inhibitor verapamil. We performed RNA sequencing (RNA-seq), whole genome sequencing (WGS), and eQTL analysis, focusing on efflux pump, transport, and secretion-associated genes. Results Of 42 ofloxacin-resistant Mtb isolates, 27 had adequate WGS coverage and acceptable RNA-seq quality. Of these 27, 7 had >2-fold reduction in ofloxacin MIC with verapamil; 6 had 2-fold reduction, and 14 had <2-fold reduction. Five genes (including Rv0191) had significantly increased expression in the MIC fold change >2 compared to <2 groups. Among regulated genes, 31 eQTLs (without ofloxacin) and 35 eQTLs (with ofloxacin) had significant allele frequency differences between MIC fold change >2 and <2 groups. Of these, Rv1410c , Rv2459 , and Rv3756c (without ofloxacin) and Rv0191 and Rv3756c (with ofloxacin) have previously been associated with antituberculosis drug resistance. Conclusions In this first reported eQTL analysis in Mtb , Rv0191 had increased gene expression and significance in eQTL analysis, making it a candidate for functional evaluation of efflux-mediated fluoroquinolone resistance in Mtb. [ABSTRACT FROM AUTHOR]

Pulmonary tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (MTB). Whole-genome sequencing (WGS) holds great promise as an advanced technology for accurately predicting anti-TB drug resistance. The development of a reliable method for detecting drug resistance is crucial in order to standardize anti-TB treatments, enhance patient prognosis, and effectively reduce the risk of transmission. In this study, our primary objective was to explore and determine the potential of WGS for assessing drug resistance based on genetic variants recommended by the World Health Organization (WHO). A total of 1105 MTB strains were selected from samples collected from 2014–2018 in Zhejiang Province, China. Phenotypic drug sensitivity tests (DST) of the anti-TB drugs were conducted for isoniazid (INH), rifampicin (RFP), streptomycin, ethambutol, fluoroquinolones (levofloxacin and moxifloxacin), amikacin, kanamycin, and capreomycin, and the drug-resistance rates were calculated. The clean WGS data of the 1105 strains were acquired and analyzed. The predictive performance of WGS was evaluated by the comparison between genotypic and phenotypic DST results. For all anti-TB drugs, WGS achieved good specificity values (>90%). The sensitivity values for INH and RFP were 91.78% and 82.26%, respectively; however, they were ≤60% for other drugs. The positive predictive values for anti-TB drugs were >80%, except for ethambutol and moxifloxacin, and the negative predictive values were >90% for all drugs. In light of the findings from our study, we draw the conclusion that WGS is a valuable tool for identifying genome-wide variants. Leveraging the genetic variants recommended by the WHO, WGS proves to be effective in detecting resistance to RFP and INH, enabling the identification of multi-drug resistant TB patients. However, it is evident that the genetic variants recommended for predicting resistance to other anti-TB drugs require further optimization and improvement. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (MTB), the causative agent of tuberculosis, stands as an immensely devastating and persistently relevant pathogen, claiming the lives of millions each year. This infectious bacterium remains a formidable global health challenge, necessitating urgent attention and comprehensive strategies to combat its profound impact on public health. MTB is a finicky bacterium that manages to sneak into macrophages and fibroblasts to avoid being eliminated. Current first-line treatments allow the control of the spread of an active MTB, but are not capable of effectively controlling when MTB is in its latent phase and struggle against MTB resistant strains. Lipid-based nanoparticles have gained significant attention in the field of tuberculosis nanotechnology treatments, owing to their compelling logical underpinnings, remarkable merits, and acknowledged demerits. These nanoparticles offer a rational approach by harnessing the unique properties of lipids, such as biocompatibility and stability, to encapsulate and protect anti-tuberculosis drugs. Their inherent ability to actively target infected macrophages holds immense promise for precise drug delivery to the infection site, enhancing therapeutic efficacy. However, it is crucial to consider potential limitations, such as the restricted payload capacity due to their small size and challenges in achieving sustained drug release. Despite these challenges, lipid-based nanotechnology represents an exciting frontier for combating drug resistance and advancing tuberculosis treatment strategies, warranting further exploration and development in this field. In addition, we emphasize the characteristics of lipid-based nanoparticles with the ability to improve the administration, stability, and dosage of these molecules. New modified systems are expected to be successful in the coming years as nanotechnology has improved various treatments in other diseases.

IntroductionNeutrophil granulocytes predominate in the lungs of patients infected with Mycobacterium tuberculosis (Mtb) in earlier stages of the disease. During infection, neutrophils release neutrophil extracellular traps (NETs), an antimicrobial mechanism by which a DNA-backbone spiked with antimicrobial components traps the mycobacteria. However, the specific mycobacterial factors driving NET formation remain unclear. Proteins from the proline-glutamic acid (PE)/proline-proline-glutamic acid (PPE) family are critical to Mtb pathophysiology and virulence.MethodsHere, we investigated NET induction by PE18, PPE26, and PE31 in primary human blood-derived neutrophils. Neutrophils were stimulated with the respective proteins for 3h, and NET formation was subsequently assessed using confocal fluorescence microscopy. Intracellular ROS levels and cell necrosis were estimated by flow cytometry. Additionally, the influence of phorbol-12-myristate-13-acetate (PMA), a known NADPH oxidase enhancer, on NET formation was examined. Neutrophil integrity following incubation with the PE/PPE proteins was evaluated using transmission electron microscopy.ResultsFor the first time, we report that stimulation of primary human blood-derived neutrophils with Mtb proteins PE18, PPE26, and PE31 resulted in the formation of NETs, which correlated with an increase in intracellular ROS levels. Notably, the presence of PMA further amplified this effect. Following incubation with the PE/PPE proteins, neutrophils were found to remain viable and structurally intact, as verified through transmission electron microscopy, indicating the occurrence of vital NET formation.DiscussionThese findings offer valuable insights that contribute to a better understanding of host-pathogen interactions during Mtb infection. Moreover, they underscore the significance of these particular Mtb antigens in triggering NET formation, representing a distinctive and previously unrecognized function of PE/PPE antigens.

Background Semiqualitative parameter SUVmax has been the most frequently used semiquantitative positron emission tomography (PET) parameter for response evaluation, but only metabolic activity of a single (most metabolic) lesion is predicted. Newer response parameters such as tumor lesion glycolysis (TLG) incorporating lesions' metabolic volume or whole-body metabolic tumor burden (MTBwb) are being explored for response evaluation. Evaluation and comparison of response with different semiquantitative PET parameters such as SUVmax and TLG in most metabolic lesion, multiple lesions (max of five), and MTBwb in advanced non-small cell lung cancer (NSCLC) patients were made. The different PET parameters were analyzed for response evaluation, overall survival (OS), and progression-free survival (PFS). Methods 18 F-FDG-PET/CT (18-fluorine-fluorodeoxyglucose positron emission tomography/computed tomography) imaging was performed in 23 patients (M = 14, F = 9, mean age = 57.6 years) with stage IIIB–IV advanced NSCLC before initiation of therapy with oral estimated glomerular filtration rate-tyrosine kinase inhibitor for early and late response evaluation. The quantitative PET parameters such as SUVmax and TLG were measured in single (most metabolic) lesion, multiple lesions, and MTBwb. The parameters SUVmax, TLG, and MTBwb were compared for early and late response evaluation and analyzed for OS and PFS Results No significant difference in change in response evaluation was seen in patients evaluated with most metabolic lesion, multiple lesions, or MTBwb. Difference in early (DC 22, NDC 1) and late (DC 20, NDC 3) response evaluation was seen that remained unchanged when lesions were measured in terms of number of lesions or the MTBwb. The early imaging was seen to be statistically significant to the OS compared with late imaging. Conclusions Single (most metabolic) lesion shows similar disease response and OS to multiple lesions and MTBwb. Response evaluation by late imaging offered no significant advantage compared with early imaging. Thus, early response evaluation with SUVmax parameter offers a good balance between clinical ease and research requisition. [ABSTRACT FROM AUTHOR]

Occurrence of non-canonical G-quadruplex (G4) DNA structures in the genome have been recognized as key factors in gene regulation and several other cellular processes. The mosR and ndhA genes involved in pathways of oxidation sensing regulation and ATP generation, respectively, make Mycobacterium tuberculosis (Mtb) bacteria responsible for oxidative stress inside host macrophage cells. Circular Dichroism spectra demonstrate stable hybrid G4 DNA conformations of mosR/ndhA DNA sequences. Real-time binding of mitoxantrone to G4 DNA with an affinity constant ~105–107 M−1, leads to hypochromism with a red shift of ~18 nm, followed by hyperchromism in the absorption spectra. The corresponding fluorescence is quenched with a red shift ~15 nm followed by an increase in intensity. A change in conformation of the G4 DNA accompanies the formation of multiple stoichiometric complexes with a dual binding mode. The external binding of mitoxantrone with a partial stacking with G-quartets and/or groove binding induces significant thermal stabilization, ~20–29 °C in ndhA/mosR G4 DNA. The interaction leads to a two/four-fold downregulation of transcriptomes of mosR/ndhA genes apart from the suppression of DNA replication by Taq polymerase enzyme, establishing the role of mitoxantrone in targeting G4 DNA, as an alternate strategy for effective anti-tuberculosis action in view of deadly multi-drug resistant tuberculosis disease causing bacterial strains t that arise from existing therapeutic treatments. [ABSTRACT FROM AUTHOR]

Abstract Background Molecular tumor boards (MTBs) match molecular alterations with targeted anticancer drugs upon failure of the available therapeutic options. Special and local needs are most likely to emerge through the comparative analysis of MTB networks, but these are rarely reported. This manuscript summarizes the state-of-art of 16 active Italian MTBs, as it emerges from an online survey curated by Alliance Against Cancer (ACC). Main text Most MTBs (13/16) are exclusively supported through local Institutional grants and meet regularly. All but one adopts a fully virtual or a mixed face-to-face/virtual calling/attendance meeting model. It appears that the ACC MTB initiative is shaping a hub-and-spoke virtual MTB network reminiscent of non-redundant, cost-effective healthcare organization models. Unfortunately, public awareness of MTB opportunities presently remains insufficient. Only one center has a website. Dedicated e-mail addresses are for the exclusive use of the MTB staff. More than half of ACC members consider a miscellanea of most or all solid and hematological malignancies, and more than one-third consider neoplasms arising at any anatomical location. The average number of Staff Members in MTBs is 9. More than 10 staff members simultaneously attend MTB meetings in 13 MTBs. A medical oncologist is invariably present and is in charge of introducing the clinical case either with (45%) or without previous discussion in organ-specific multidisciplinary Boards. All but two MTBs take charge of not only patients with no standard-of-care (SoC) therapy option, but also cases receiving NGS profiling in SoC settings, implying a larger number of yearly cases. All MTBs run targeted NGS panels. Three run whole-exome and/or RNAseq approaches. ESCAT-ESMO and/or Onco-KB levels of evidence are similarly used for diagnostic reporting. Most MTBs (11) provide a written diagnostic report within 15 days. Conclusions are invariably communicated to the patient by the medical oncologist. Conclusions MTB networking is crucial not only for molecular diagnosis and therapy assignment, but also for healthcare governance. Survey results show that MTBs review therapeutic opportunities at the crossover between standard-of-care with off-label, the former task being much beyond their scope. Societal and scientific implications of this beyond-the-scope MTB function may be relevant for healthcare in Italy and abroad.

Tuberculosis is a worldwide contagion caused by Mycobacterium tuberculosis (MTB). MTB is characterized by intracellular parasitism and is semi-dormant inside host cells. The persistent inflammation caused by MTB can form a granuloma in lesion regions and intensify the latency of bacteria. In recent years, several studies have proven that long non-coding RNAs (lncRNAs) play critical roles in modulating autophagy. In our study, the Gene Expression Omnibus (GEO) databases were searched for lncRNAs that are associated with tuberculosis. We found that lncRNA differentiation antagonizing non-protein coding RNA (DANCR) increased in the peripheral blood samples collected from 54 pulmonary tuberculosis patients compared to 23 healthy donors. By constructing DANCR overexpression cells, we analyzed the possible cellular function of DANCR. After analyzing our experiments, it was found that the data revealed that upregulation of DANCR facilitated the expression of signal transducer and activator of transcription 3, autophagy-related 4D cysteine peptides, autophagy-related 5, Ras homolog enriched in the brain, and microtubule-associated protein 1A/1B light chain 3 (STAT3, ATG4D, ATG5, RHEB, and LC3, respectively) by sponging miR-1301-3p and miR-5194. Immunofluorescence analysis indicated that DANCR played a positive role in both autophagosome formation and fusion of autolysosomes in macrophages. The colony-forming unit (CFU) assay data also showed that the cells overexpressing DANCR were more efficient in eliminating the intracellular H37Ra strain. Consequently, these data suggest that DANCR restrained intracellular survival of M. tuberculosis by promoting autophagy via miR-1301-3p and miR-5194. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is one of the leading causes of infectious mortality from a single infectious agent, Mycobacterium tuberculosis (MTB). This study evaluated the performance of the newly developed BZ TB/NTM NALF assay, which integrated loop-mediated isothermal amplification and lateral flow immunochromatographic assay technologies, for the detection of MTB. A total of 80 MTB-positive samples and 115 MTB-negative samples were collected, all of which were confirmed by TB real-time PCR (RT-PCR) using either AdvanSureTM TB/NTM RT-PCR Kit or Xpert® MTB/RIF Assay. The performance of the BZ TB/NTM NALF assay was evaluated by calculating its sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) in comparison to those of the RT-PCR methods. Compared to the RT-PCR, the sensitivity, specificity, PPV, and NPV of BZ TB/NTM NALF assay were 98.7%, 99.1%, 98.7%, and 99.1%, respectively. The concordance rate between BZ TB/NTM NALF and RT-PCR was 99.0%. Rapid and simple detection of MTB is essential for global case detection and further elimination of TB. The performance of the BZ TB/NTM NALF Assay is acceptable with a high concordance with RT-PCR, indicating that it is reliable for use in a low-resource environment. [ABSTRACT FROM AUTHOR]

SUMMARY: Plants colonized the land approximately 470 million years ago, coinciding with the development of apical cells that divide in three planes. The molecular mechanisms that underly the development of the 3D growth pattern are poorly understood, mainly because 3D growth in seed plants starts during embryo development. In contrast, the transition from 2D to 3D growth in the moss Physcomitrium patens has been widely studied, and it involves a large turnover of the transcriptome to allow the establishment of stage‐specific transcripts that facilitate this developmental transition. N6‐Methyladenosine (m6A) is the most abundant, dynamic and conserved internal nucleotide modification present on eukaryotic mRNA and serves as a layer of post‐transcriptional regulation directly affecting several cellular processes and developmental pathways in many organisms. In Arabidopsis, m6A has been reported to be essential for organ growth and determination, embryo development and responses to environmental signals. In this study, we identified the main genes of the m6A methyltransferase complex (MTC), MTA, MTB and FIP37, in P. patens and demonstrate that their inactivation leads to the loss of m6A in mRNA, a delay in the formation of gametophore buds and defects in spore development. Genome‐wide analysis revealed several transcripts affected in the Ppmta background. We demonstrate that the PpAPB1–PpAPB4 transcripts, encoding central factors orchestrating the transition from 2D to 3D growth in P. patens, are modified by m6A, whereas in the Ppmta mutant the lack of the m6A marker is associated with a corresponding decrease in transcript accumulation. Overall, we suggest that m6A is essential to enable the proper accumulation of these and other bud‐specific transcripts directing the turnover of stage‐specific transcriptomes, and thus promoting the transition from protonema to gametophore buds in P. patens. Significance Statement: The developmental transition from 2D to 3D growth in Physcomitrium patens involves transcriptome rearrangements. The N6‐methyladenosine (m6A) modification of mRNA is a post‐transcriptional regulator affecting cellular and developmental pathways in eukaryotes. The inactivation of genes encoding for m6A methyltransferase complex proteins leads to developmental defects in gametophore bud and spore. The PpAPB1–PpAPB4 transcripts, encoding for central regulators of bud formation, are modified by m6A, among many others, suggesting that m6A enables the proper accumulation of specific transcripts, promoting bud development. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) is a serious infectious disease caused by Mycobacterium tuberculosis (MTB) and a significant health concern worldwide. The main threat to the elimination of TB is the development of resistance by MTB to the currently used antibiotics and more extended treatment methods, which is a massive burden on the health care system. As a result, there is an urgent need to identify new, effective therapeutic strategies with fewer adverse effects. The traditional medicines found in South Asia and Africa have a reservoir of medicinal plants and plant-based compounds that are considered another reliable option for human beings to treat various diseases. Abundant research is available for the biotherapeutic potential of naturally occurring compounds in various diseases but has been lagging in the area of TB. Plant-based compounds, or phytoproducts, are being investigated as potential anti-mycobacterial agents by reducing bacterial burden or modulating the immune system, thereby minimizing adverse effects. The efficacy of these phytochemicals has been evaluated through drug delivery using nanoformulations. This review aims to emphasize the value of anti-TB compounds derived from plants and provide a summary of current research on phytochemicals with potential anti-mycobacterial activity against MTB. This article aims to inform readers about the numerous potential herbal treatment options available for combatting TB. [ABSTRACT FROM AUTHOR]

N6-adenosine methylation (m6A) is a prevalent form of RNA modification found in the expressed transcripts of many eukaryotic organisms. Moreover, m6A methylation is a dynamic and reversible process that requires the functioning of various proteins and their complexes that are evolutionarily conserved between species and include methylases, demethylases, and m6A-binding proteins. Over the past decade, the m6A methylation process in plants has been extensively studied and the understanding thereof has drastically increased, although the regulatory function of some components relies on information derived from animal systems. Notably, m6A has been found to be involved in a variety of factors in RNA processing, such as RNA stability, alternative polyadenylation, and miRNA regulation. The circadian clock in plants is a molecular timekeeping system that regulates the daily and rhythmic activity of many cellular and physiological processes in response to environmental changes such as the day-night cycle. The circadian clock regulates the rhythmic expression of genes through post-transcriptional regulation of mRNA. Recently, m6A methylation has emerged as an additional layer of post-transcriptional regulation that is necessary for the proper functioning of the plant circadian clock. In this review, we have compiled and summarized recent insights into the molecular mechanisms behind m6A modification and its various roles in the regulation of RNA. We discuss the potential role of m6A modification in regulating the plant circadian clock and outline potential future directions for the study of mRNA methylation in plants. A deeper understanding of the mechanism of m6A RNA regulation and its role in plant circadian clocks will contribute to a greater understanding of the plant circadian clock. [ABSTRACT FROM AUTHOR]

Infection with Mycobacterium tuberculosis (Mtb) results in the primary formation of a densely packed inflammatory foci that limits entry of therapeutic agents into pulmonary sites where organisms reside. No current therapeutic regimens exist that modulate host immune responses to permit increased drug penetration to regions of pathological damage during tuberculosis disease. Lactoferrin is a natural iron-binding protein previously demonstrated to modulate inflammation and granuloma cohesiveness, while maintaining control of pathogenic burden. Studies were designed to examine recombinant human lactoferrin (rHLF) to modulate histological progression of Mtb-induced pathology in a non-necrotic model using C57Bl/6 mice. The rHLF was oral administered at times corresponding to initiation of primary granulomatous response, or during granuloma maintenance. Treatment with rHLF demonstrated significant reduction in size of primary inflammatory foci following Mtb challenge, and permitted penetration of ofloxacin fluoroquinolone therapeutic to sites of pathological disruption where activated (foamy) macrophages reside. Increased drug penetration was accompanied by retention of endothelial cell integrity. Immunohistochemistry revealed altered patterns of M1-like and M2-like phenotypic cell localization post infectious challenge, with increased presence of M2-like markers found evenly distributed throughout regions of pulmonary inflammatory foci in rHLF-treated mice. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (Mtb) is a smart and successful pathogen since it can persist in the intimidating environment of the host by taming and tuning the immune system. Mtb releases MPT64 (Rv1980c) protein in high amounts in patients with active tuberculosis (TB). Consequently, we were curious to decipher the role of MPT64 on the differentiating dendritic cells (DCs) and its relation to evading the immune system. We observed that pre-exposure of differentiating DCs to MPT64 (DCMPT64) transformed them into a phenotype of myeloid-derived suppressor cells (MDSCs). DCMPT64 expressed a high level of immunosuppressive molecules PD-L1, TIM-3, nitric oxide (NO), arginase 1, IDO-1, IL-10 and TGF-β, but inhibited the production of pro-inflammatory cytokines TNF-α, IL-6 and IL-12. DCMPT64 chemotaxis function was diminished due to the reduced expression of CCR7. DCMPT64 promoted the generation of regulatory T cells (Tregs) but inhibited the differentiation of Th1 cells and Th17 cells. Further, high lipid and methylglyoxal content, and reduced glucose consumption by DCMPT64, rendered them metabolically quiescent and consequently, reduced DCMPT64 ability to phagocytose Mtb and provided a safer shelter for the intracellular survival of the mycobacterium. The mechanism identified in impairing the function of DCMPT64 was through the increased production and accumulation of methylglyoxal. Hence, for the first time, we demonstrate the novel role of MPT64 in promoting the generation of MDSCs to favor Mtb survival and escape its destruction by the immune system. [ABSTRACT FROM AUTHOR]