Transcriptomic profile of mycobacterium smegmatis in response to an imidazo[1,2-b][1,2,4,5]tetrazine reveals its possible impact on iron metabolism

Tuberculosis (TB), caused by the Mycobacterium tuberculosis complex bacteria, is one of the most pressing health problems. The development of new drugs and new therapeutic regimens effective against the pathogen is one of the greatest challenges in the way of tuberculosis control. Imidazo[1,2-b][1,2,4,5]tetrazines have shown promising activity against M. tuberculosis and M. smegmatis strains. Mutations in MSMEG_1380 lead to mmpS5–mmpL5 operon overexpression, which provides M. smegmatis with efflux-mediated resistance to imidazo[1,2-b][1,2,4,5]tetrazines, but the exact mechanism of action of these compounds remains unknown. To assess the mode of action of imidazo[1,2-b][1,2,4,5]tetrazines, we analyzed the transcriptomic response of M. smegmatis to three different concentrations of 3a compound: 1/8×, 1/4×, and 1/2× MIC. Six groups of genes responsible for siderophore synthesis and transport were upregulated in a dose-dependent manner, while virtual docking revealed proteins involved in siderophore synthesis as possible targets for 3a

Questionnaire-assessed risk of sleep apnea in inpatients with various endocrine disorders

Background: Sleep breathing disorders can be an additional risk factor for the development of cardiovascular disorders in patients with endocrine disorders.Aim: To assess the sleep apnea risk in patients with various endocrine disorders undergoing inpatient treatment.Materials and methods: The sleep apnea risk and the severity of daytime sleepiness were evaluated in 282 inpatients with endocrine disorders based on the following questionnaires: the Epworth Sleepiness Scale (ESS), the sleep apnea screening questionnaire (SAS), and the Berlin sleep apnea risk questionnaire (BQ). To identify the real prevalence of sleep respiratory disorders in endocrine patients we performed cardio-respiratory monitoring with Watch-PAT200 (“Itamar Medical”, Israel) device in 81 patients.Results: A high sleep apnea risk according to the BQ was detected in 59.9% (160/267) of the patients, according to the SAS in 53.53% (144/269), excessive daytime sleepiness (≥ 11 points) was found in 21.66% (60/277) of the patients with the ESS. Among the patients undergoing cardiorespiratory monitoring, sleep apnea was detected in 84% (68/81), including severe apnea in 38.2% (26/68). The highest sleep apnea risk by BQ and SAS was observed in the patients with type 2 diabetes, acromegaly and hypercortisolism. Excessive daytime sleepiness by the ESS was most noticeable in those with hypercortisolism and thyrotoxicosis.Conclusion: The high risk of sleep apnea in the inpatients with type 2 diabetes mellitus, acromegaly, hypercorticism, and hypothyroidism makes it necessary to include its active screening into the algorithm of their inpatient assessment to rule out any sleep breathing disorder

Assessment of muscle and fat mass in type 2 diabetes mellitus patients by dual-energy X-ray absorptiometry

Background: Obesity is an important health problem, as its prevalence has reached an epidemic level and continues to increase steadily resulting in higher risk of cardiovascular diseases and metabolic disorders. Currently, new methods and criteria are being developed to assess fat and muscle mass, as well as criteria for diagnosing obesity and sarcopenia.Aim: To assess the quantitative composition of muscle and adipose tissue in type 2 diabetes mellitus patients based on the dual-energy X-ray absorptiometry for the diagnosis of obesity and sarcopenia.Materials and methods: We examined 42 type 2 diabetic in-patients admitted to the Department of Therapeutic Endocrinology. Dual-energy X-ray absorptiometry was performed in all patients with subsequent assessment of the composition of muscle and fat tissue.Results: If assessed by the body mass index, all patients had an excess body weight: median, 32.25 [29.75; 35.70]; in men, 31.3 [28.19; 34.63], in women, 32.29 [30.26; 36.54]. 26.2% of the patients (11/42) were overweight, but not obese. Female patients had more severe obesity than male (in total, 33.3% (10/30) of women had 2nd and 3rd degree of obesity, while men 16.7% (2/12)). The assessment by the fat mass index (FMI) showed that 2.4% (1/42) of the patients were normal-weight. Median FMI was 11.91 [10.40; 13.78] (in men, 8.86 [7.46; 12.1], in women, 12.35 [11.55; 15.47]). Overweight was found in 52.4% (22/42) of the patients; in total, 2nd and 3rd degree of obesity was observed in 25% (3/12) of the men and only in 6.6% (2/30) of the women. Median Appendicular Lean Mass Index (ALMI) in the total group was 7.99 [7.32; 9.05], being expectedly higher than in women: 9.19 [8.42; 9.45] and 7.58 [7.24; 8.49], respectively. Median T-score ALMI was 2.32 [1.73; 3.08], Z-score ALMI 2.15 [1.47; 3.54]. In general, there was a decrease in the appendicular muscle mass with age. There was an inverse correlation between the age and T-score ALMI (r = -0.319, р = 0.020), as well as between the age and Z-score ALMI (r = -0.634, p = 0.000). According to the results of T-score ALMI and Z-score ALMI, there were no patients with sarcopenia. However, the calculation of the T- and Z-criteria, corrected for fat mass, has led to a significant decrease of the medians of these parameters and allowed to identify a group of patients meeting the criteria of sarcopenia (97.6%, 41/42).Conclusion: Based on ALMI, T-ALMI, and Z-ALMI, there were no patients with sarcopenia. After these criteria were corrected for fat mass, the number of such patients increased to 97.6% (41/42) and 85.7% (36/42), respectively. The potential use of the adjusted T-ALMI (FMI) and Z-ALMI (FMI) as criteria for sarcopenia and muscle mass reduction compared to the age-related normal values, as well as the classification of obesity by FMI should be studied in large epidemiological studies in different populations

Identification of Mutations Conferring Tryptanthrin Resistance to Mycobacterium smegmatis

Tuberculosis (TB), caused by Mycobacterium tuberculosis, is a global burden, responsible for over 1 million deaths annually. The emergence and spread of drug-resistant M. tuberculosis strains (MDR-, XDR- and TDR-TB) is the main challenge in global TB-control, requiring the development of novel drugs acting on new biotargets, thus able to overcome the drug-resistance. Tryptanthrin is a natural alkaloid, with great therapeutic potential due to its simple way of synthesis and wide spectrum of biological activities including high bactericidal activity on both drug-susceptible and MDR M. tuberculosis strains. InhA was suggested as the target of tryptanthrins by in silico modeling, making it a promising alternative to isoniazid, able to overcome drug resistance provided by katG mutations. However, neither the mechanism of action of tryptanthrin nor the mechanism of resistance to tryptanthrins was ever confirmed in vitro. We show that the MmpS5-MmpL5 efflux system is able to provide resistance to tryptanthrins using an in-house test-system. Comparative genomic analysis of spontaneous tryptanthrin-resistant M. smegmatis mutants showed that mutations in MSMEG_1963 (EmbR transcriptional regulator) lead to a high-level resistance, while those in MSMEG_5597 (TetR transcriptional regulator) to a low-level one. Mutations in an MFS transporter gene (MSMEG_4427) were also observed, which might be involved in providing a basal level of tryptanthrins-resistance

The Gene Expression Profile Differs in Growth Phases of the Bifidobacterium Longum Culture

To date, transcriptomics have been widely and successfully employed to study gene expression in different cell growth phases of bacteria. Since bifidobacteria represent a major component of the gut microbiota of a healthy human that is associated with numerous health benefits for the host, it is important to study them using transcriptomics. In this study, we applied the RNA-Seq technique to study global gene expression of B. longum at different growth phases in order to better understand the response of bifidobacterial cells to the specific conditions of the human gut. We have shown that in the lag phase, ABC transporters, whose function may be linked to active substrate utilization, are increasingly expressed due to preparation for cell division. In the exponential phase, the functions of activated genes include synthesis of amino acids (alanine and arginine), energy metabolism (glycolysis/gluconeogenesis and nitrogen metabolism), and translation, all of which promote active cell division, leading to exponential growth of the culture. In the stationary phase, we observed a decrease in the expression of genes involved in the control of the rate of cell division and an increase in the expression of genes involved in defense-related metabolic pathways. We surmise that the latter ensures cell survival in the nutrient-deprived conditions of the stationary growth phase

Metagenomic Profiles of the Intestinal Virome of Long-Livers

The microbial community of the human intestine is important for maintaining human health. It has been reported that the gut microbiome changes with age, and it can be enrichedwith certain beneficial bacteria while also losing certain commensal bacteria.Little is known about the gut virome of long-livers. Our research aimed to extract, sequence and analyze the viral fraction of long-livers’ gut microbiota in comparison with those of young adults and the elderly. We were thereby able to characterize the gut virome profiles and viral diversity of three age groups. Keywords: aging, gut microbiome, viral metagenomics, bacteriophage

Metagenomic Profiles of the Intestinal Virome of Long-Livers

The microbial community of the human intestine is important for maintaining human health. It has been reported that the gut microbiome changes with age, and it can be enrichedwith certain beneficial bacteria while also losing certain commensal bacteria.Little is known about the gut virome of long-livers. Our research aimed to extract, sequence and analyze the viral fraction of long-livers' gut microbiota in comparison with those of young adults and the elderly. We were thereby able to characterize the gut virome profiles and viral diversity of three age groups. Keywords: aging, gut microbiome, viral metagenomics, bacteriophage

Upregulation of NETO2 gene in colorectal cancer

Abstract Background Neuropilin and tolloid-like 2 (NETO2) is a single-pass transmembrane protein that has been shown primarily implicated in neuron-specific processes. Upregulation of NETO2 gene was also detected in several cancer types. In colorectal cancer (CRC), it was associated with tumor progression, invasion, and metastasis, and seems to be involved in epithelial-mesenchymal transition (EMT). However, the mechanism of NETO2 action is still poorly understood. Results We have revealed significant increase in the expression of NETO2 gene and deregulation of eight EMT-related genes in CRC. Four of them were upregulated (TWIST1, SNAIL1, LEF1, and FOXA2); the mRNA levels of other genes (FOXA1, BMP2, BMP5, and SMAD7) were decreased. Expression of NETO2 gene was weakly correlated with that of genes involved in the EMT process. Conclusions We found considerable NETO2 upregulation, but no significant correlation between the expression of NETO2 and EMT-related genes in CRC. Thus, NETO2 may be involved in CRC progression, but is not directly associated with EMT

HK3 overexpression associated with epithelial-mesenchymal transition in colorectal cancer

Abstract Background Colorectal cancer (CRC) is a common cancer worldwide. The main cause of death in CRC includes tumor progression and metastasis. At molecular level, these processes may be triggered by epithelial-mesenchymal transition (EMT) and necessitates specific alterations in cell metabolism. Although several EMT-related metabolic changes have been described in CRC, the mechanism is still poorly understood. Results Using CrossHub software, we analyzed RNA-Seq expression profile data of CRC derived from The Cancer Genome Atlas (TCGA) project. Correlation analysis between the change in the expression of genes involved in glycolysis and EMT was performed. We obtained the set of genes with significant correlation coefficients, which included 21 EMT-related genes and a single glycolytic gene, HK3. The mRNA level of these genes was measured in 78 paired colorectal cancer samples by quantitative polymerase chain reaction (qPCR). Upregulation of HK3 and deregulation of 11 genes (COL1A1, TWIST1, NFATC1, GLIPR2, SFPR1, FLNA, GREM1, SFRP2, ZEB2, SPP1, and RARRES1) involved in EMT were found. The results of correlation study showed that the expression of HK3 demonstrated a strong correlation with 7 of the 21 examined genes (ZEB2, GREM1, TGFB3, TGFB1, SNAI2, TWIST1, and COL1A1) in CRC. Conclusions Upregulation of HK3 is associated with EMT in CRC and may be a crucial metabolic adaptation for rapid proliferation, survival, and metastases of CRC cells