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Herpesvirus (HV) is widely distributed among cetacean populations, with the highest prevalence reported in the Mediterranean Sea. In this study, a comprehensive analysis was conducted, including epidemiological, phylogenetic, and pathological aspects, with particular emphasis on neuropathology, to better understand the impact of HV in these animals. Our results show a higher presence of HV in males compared to females, with males exhibiting a greater number of positive tissues. Additionally, adults were more frequently affected by HV infection than juveniles, with no infections detected in calves or neonates. The affected species were striped (Stenella coeruleoalba) and bottlenose dolphins (Tursiops truncatus). The highest positivity rates were observed in the genital system, cerebrum, and skin tissues. Phylogenetic analysis indicated a higher occurrence of Gammaherpesvirus (GHV) sequences but increased genetic diversity within Alphaherpesvirus (AHV). Key neuropathological features included astro-microgliosis (n = 4) and meningitis with minimal to mild perivascular cuffing (n = 2). The presence of concurrent infections with other pathogens, particularly cetacean morbillivirus (CeMV), underscores the complex nature of infectious diseases in cetaceans. However, the presence of lesions at the Central Nervous System (CNS) with molecular positivity for GHV, excluding the involvement of other potential neurotropic agents, would confirm the potential of this HV subfamily to induce neurological damage. Pathological examination identified lesions in other organs that could potentially be associated with HV, characterized by lymphoid depletion and tissue inflammation. These findings enhance our understanding of HV in odontocetes and highlight the need for ongoing research into the factors driving these infections and their broader implications. [ABSTRACT FROM AUTHOR]

Tuberculous meningitis (TBM) is a serious form of TB disease that can result in high morbidity and mortality, particularly if there are delays in diagnosis and treatment. In this case report, a 9‐month‐old girl was admitted with persistent vomiting and focal seizures. On examination, she was found to have a right‐side hemiparesis. Brain imaging showed intense nodular leptomeningeal enhancement, hydrocephalus, a hypolucent lesion in the left basal ganglia, arterial stenosis and vasculitis, and an old ischemic insult. The patient was initially diagnosed with an acute ischemic stroke and was treated with aspirin and antiepileptic drugs. The patient's condition failed to improve despite initial treatment, leading to further diagnostic procedures. The results uncovered a diagnosis of TBM. The case highlights the importance of considering TBM as a possible cause of neurological symptoms, especially during the coronavirus disease 2019 (COVID‐19) pandemic where similar symptoms can be present in cases of neurological complications of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection and multisystem inflammatory syndrome in children (MIS‐C). [ABSTRACT FROM AUTHOR]

In recent years, the Internet of medical things (IoMT) has become a significant technological advancement in the healthcare sector. This systematic review aims to identify and summarize the various applications, key challenges, and proposed technical solutions within this domain, based on a comprehensive analysis of the existing literature. This review highlights diverse applications of the IoMT, including mobile health (mHealth) applications, remote biomarker detection, hybrid RFID-IoT solutions for scrub distribution in operating rooms, IoT-based disease prediction using machine learning, and the efficient sharing of personal health records through searchable symmetric encryption, blockchain, and IPFS. Other notable applications include remote healthcare management systems, non-invasive real-time blood glucose measurement devices, distributed ledger technology (DLT) platforms, ultra-wideband (UWB) radar systems, IoT-based pulse oximeters, accident and emergency informatics (A&EI), and integrated wearable smart patches. The key challenges identified include privacy protection, sustainable power sources, sensor intelligence, human adaptation to sensors, data speed, device reliability, and storage efficiency. The proposed mitigations encompass network control, cryptography, edge-fog computing, and blockchain, alongside rigorous risk planning. The review also identifies trends and advancements in the IoMT architecture, remote monitoring innovations, the integration of machine learning and AI, and enhanced security measures. This review makes several novel contributions compared to the existing literature, including (1) a comprehensive categorization of IoMT applications, extending beyond the traditional use cases to include emerging technologies such as UWB radar systems and DLT platforms; (2) an in-depth analysis of the integration of machine learning and AI in IoMT, highlighting innovative approaches in disease prediction and remote monitoring; (3) a detailed examination of privacy and security measures, proposing advanced cryptographic solutions and blockchain implementations to enhance data protection; and (4) the identification of future research directions, providing a roadmap for addressing current limitations and advancing the scientific understanding of IoMT in healthcare. By addressing current limitations and suggesting future research directions, this work aims to advance scientific understanding of the IoMT in healthcare. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis is the causative agent of tuberculosis. M. tuberculosis can survive in a dormant state within the granuloma, avoiding the host-mounting immune attack. M. tuberculosis bacilli in this state show increased tolerance to antibiotics and stress conditions, and thus the transition of M. tuberculosis to the nonreplicating dormant state acts as an obstacle to tuberculosis treatment. M. tuberculosis in the granuloma encounters hostile environments such as hypoxia, nitric oxide, reactive oxygen species, low pH, and nutrient deprivation, etc., which are expected to inhibit respiration of M. tuberculosis. To adapt to and survive in respiration-inhibitory conditions, it is required for M. tuberculosis to reprogram its metabolism and physiology. In order to get clues to the mechanism underlying the entry of M. tuberculosis to the dormant state, it is important to understand the mycobacterial regulatory systems that are involved in the regulation of gene expression in response to respiration inhibition. In this review, we briefly summarize the information regarding the regulatory systems implicated in upregulation of gene expression in mycobacteria exposed to respiration-inhibitory conditions. The regulatory systems covered in this review encompass the DosSR (DevSR) two-component system, SigF partner switching system, MprBA-SigE-SigB signaling pathway, cAMP receptor protein, and stringent response., (© 2023. The Author(s), under exclusive licence to Microbiological Society of Korea.)

Parasite expression libraries have so far been screened with antibodies, DNA probes or T cell clones. Immunity to many parasites, such as Mycobacterium leprae, is largely mediated by T cells, and so the screening of such libraries for T cell epitopes is an important step toward the development of effective vaccines and diagnostic reagents. A new method for screening of lambda gt11 libraries with uncloned T cell populations is presented here, which takes advantage of the fact that the recombinant proteins contain beta-galactosidase as their leader peptide; this allows them to be semipurified by means of anti-beta-galactosidase antibodies coated on the bottom of microtiter plate wells, within which a proliferation assay can then be carried out. Optimum conditions for the assay were determined, using the M. leprae 18-kDa antigen as a test antigen.

Asthma is a prevalent chronic disease, contributing significantly to the global burden of disease and economic costs. Despite advances in treatment, inadequate disease management and reliance on reliever medications lead to preventable deaths. Telemedicine, defined as the use of information and communication technology to improve healthcare access, has gained global attention, especially during the COVID-19 pandemic. This systematic review examines the effectiveness of home monitoring systems in managing severe asthma. A systematic literature search was conducted in PubMed, Web of Science, Scopus, and Cochrane Library, focusing on studies from 2014 to 2024. Fourteen studies involving 9093 patients were analyzed. The results indicate that telemedicine, through tools such as mobile applications and portable spirometers, positively impacts asthma control, self-management, and quality of life. Home spirometry, in particular, shows strong agreement with clinic spirometry, offering a feasible alternative for continuous monitoring. Digital coaching and machine learning-based telemedicine applications also demonstrate significant potential in improving asthma outcomes. However, challenges such as technology accessibility, data privacy, and the need for standardized protocols remain. This review highlights the promise of telemedicine in asthma management and calls for further research to optimize its implementation and address existing barriers. [ABSTRACT FROM AUTHOR]

Introduction: Asthma is a common disease with a global burden of 358 million patients. Despite improvements in pharmacological and non-pharmacological treatments, many patients still do not achieve complete asthma control. Therefore, innovative pharmacotherapy is important. Areas covered: Following a semi-structured search in Pubmed, an overview of advances in inhaled asthma therapy is provided, looking at innovations in digital inhalers, eco-friendly inhalers and novel inhaled biologic therapies, antibiotics and vaccines, as well as other potential novel asthma therapy targets. Expert opinion: Digital inhalers, sending reminders and monitoring inhalation technique electronically, can support medication adherence and improve asthma control. To reduce the global warming potential of traditional aerosols used in pressurized metered-dose inhalers (HFA-134a, HFA-227ea), greener alternatives are under development (HFA-152a, HFO-1234ze) that are expected to be available by 2025. Current pharmacological advances in asthma therapy are mainly achieved by novel biologicals (anti-IgE, anti-IL5, anti-IL4/13, and anti-TSLP) targeting specific severe asthma phenotypes. While injection is the usual administration route for biologics and vaccines used in asthma, inhalation is an option being explored, although several (mainly formulation) challenges need to be overcome. Other potential novel future inhaled asthma therapies include anti-IL-33/ST2 biologicals and JAK inhibitors, all still requiring more clinical evidence. [ABSTRACT FROM AUTHOR]

Background: Leveraging well-established DNA-level drug resistance mechanisms, whole-genome sequencing (WGS) has emerged as a valuable methodology for predicting drug resistance. As the most effective second-line anti-tuberculosis (anti-TB) drugs, fluoroquinoloness (FQs) are generally used to treat multidrug-resistant tuberculosis (MDR-TB, defined as being resistant to resistant to rifampicin and isoniazid) or rifampicin-resistant tuberculosis (RR-TB). However, FQs are also commonly used in the management of other bacterial infections. There are few published data on the rates of FQs resistance among rifampicin-susceptible TB. The prevalence of FQs resistance among TB patients who are rifampicin-susceptible has not been studied in Zhejiang Province, China. The goal of this study was to provide a baseline characterization of the prevalence of FQs resistance, particularly among rifampicin-susceptible TB in Zhejiang Province, China. Methods: Based on WGS, we have investigated the prevalence of FQs resistance among rifampicin-susceptible TB in Zhejiang Province. All pulmonary TB patients with positive cultures who were identified in Zhejiang area during TB drug resistance surveillance from 2018 to 2019 have enrolled in this populationbased retrospective study. Results: The rate of FQs resistance was 4.6% (32/698) among TB, 4.0% (27/676) among rifampicin-susceptible TB, and 22.7% (5/22) among RR-TB. According to WGS, strains that differ within 12 single-nucleotide polymorphisms (SNPs) were considered to be transmission of FQ-resistant strains. Specifically, 3.7% (1/27) of FQs resistance was caused by the transmission of FQs-resistant strains among the rifampicin-susceptible TB and 40.7% (11/27) of FQs resistance was identified as hetero-resistance. Conclusion: The prevalence of FQs resistance among TB patients who were rifampicin-susceptible was severe in Zhejiang. The emergence of FQs resistance in TB isolates that are rifampicin-susceptible was mainly caused by the selection of drug-resistant strains. In order to prevent the emergence of FQs resistance, the WGS-based surveillance system for TB should be urgently established, and clinical awareness of the responsible use of FQs for respiratory infections should be enhanced. [ABSTRACT FROM AUTHOR]

SigE is one of the main regulators of mycobacterial stress response and is characterized by a complex regulatory network based on two pathways, which have been partially characterized in conditions of surface stress. The first pathway is based on the induction of sigE transcription by the two-component system MprAB, while the second is based on the degradation of SigE antisigma factor RseA by ClpC1P2, a protease whose structural genes are induced by ClgR. We characterized the dynamics of the SigE network activation in conditions of surface stress and low pH in Mycobacterium tuberculosis. Using a series of mutants in which the main regulatory nodes of the network have been inactivated, we could explore their hierarchy, and we determined that MprAB had a key role in the network activation in both stress conditions through the induction of sigE. However, while in conditions of surface stress the absence of MprAB totally abrogated sigE induction, under low pH conditions it only resulted in a small delay of the induction of sigE. In this case, sigE induction was due to SigH, which acted as a MprAB backup system. The ClgR pathway, leading to the degradation of the SigE anti-sigma factor RseA, was shown to be essential for the activation of the SigE network only following surface stress, where it showed an equal hierarchy with the MprAB pathway. [ABSTRACT FROM AUTHOR]

Tuberculosis (TB) remains a persistent epidemic, and the emergence of drug-resistant Mycobacterium tuberculosis (Mtb) presents a global healthcare threat. While some new agents have been successfully introduced, innovative technologies to evaluate emerging anti-TB compounds are required to inform transformative approaches. Mtb is an obligate human pathogen, and consequently utilizing models that are consistent with human disease is likely to be critical. We have developed a human 3-dimensional (3-D) cell culture model that reflects human disease gene expression patterns and causes Mtb to become pyrazinamide sensitive in vitro. Here, we identify key differences in virulence between the standard laboratory strain, Mtb H37Rv, and clinical isolates. We demonstrate that Mtb H37Rv is attenuated in the 3-D system, more susceptible to antibiotics and hyperinflammatory compared to clinical isolates. Prolonged in vitro culture of a clinical strain leads to attenuation. We then characterise antibiotic sensitivity of multi-drug-resistant Mtb within the 3-D model and define relative bactericidal activity. Finally, we demonstrate that verapamil increases efficacy of bedaquiline and delamanid antibiotic therapy. Taken together, our findings suggest that studying virulent clinical strains in an advanced cell culture system is a powerful adjunct to established methodologies to evaluate new interventions for TB. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (Mtb), the bacterium that causes tuberculosis (TB), can be difficult to treat because of drug tolerance. Increased intracellular polyphosphate (polyP) in Mtb enhances tolerance to antibiotics, and capsular polyP in Neisseria gonorrhoeae potentiates resistance to antimicrobials. The mechanism by which bacteria utilize polyP to adapt to antimicrobial pressure is not known. In this study, we found that Mtb adapts to the TB frontline antibiotic isoniazid (INH) by enhancing the accumulation of cellular, extracellular, and cell surface polyP. Gallein, a broad-spectrum inhibitor of the polyphosphate kinase that synthesizes polyP, prevents this INH-induced increase in extracellular and cell surface polyP levels. Gallein and INH work synergistically to attenuate Mtb's ability to grow in in vitro culture and within human macrophages. Mtb when exposed to INH, and in the presence of INH, gallein inhibits cell envelope formation in most but not all Mtb cells. Metabolomics indicated that INH or gallein have a modest impact on levels of Mtb metabolites, but when used in combination, they significantly reduce levels of metabolites involved in cell envelope synthesis and amino acid, carbohydrate, and nucleoside metabolism, revealing a synergistic effect. These data suggest that gallein represents a promising avenue to potentiate the treatment of TB. [ABSTRACT FROM AUTHOR]

Introduction: Mycobacterium tuberculosis (Mtb), the main cause of tuberculosis (TB), has brought a great burden to the world's public health. With the widespread use of Mtb drug-resistant strains, the pressure on anti-TB treatment is increasing. Anti-TB drugs with novel structures and targets are urgently needed. Previous studies have revealed a series of CYPs with important roles in the survival and metabolism of Mtb. However, there is little research on the structure and function of CYP138. Methods: In our study, to discover the function and targetability of CYP138, a cyp138-knockout strain was built, and the function of CYP138 was speculated by the comparison between cyp138-knockout and wild-type strains through growth curves, growth status under different carbon sources, infection curves, SEM, MIC tests, quantitative proteomics, and lipidomics. Results and discussion: The knockout of cyp138 was proven to affect the Mtb's macrophage infection, antibiotics susceptibility, and the levels of fatty acid metabolism, membrane-related proteins, and lipids such as triacylglycerol. We proposed that CYP138 plays an important role in the synthesis and decomposition of lipids related to the cell membrane structure as a new potential anti-tuberculosis drug target. [ABSTRACT FROM AUTHOR]

Background: It was previously shown that GlnA3sc enabled Streptomyces coelicolor to survive in excess polyamines. However, subsequent studies revealed that Rv1878, the corresponding Mycobacterium tuberculosis (M.tb) ortholog, was not essential for the detoxification of spermine (Spm), in M.tb. On the other hand, the multi-drug efflux pump Rv1877 was previously shown to enable export of a wide range of compounds, while Rv0191 was shown to be more specific to chloramphenicol. Rationale: Therefore, we first wanted to determine if detoxification of Spm by efflux can be achieved by any efflux pump, or if that was dependent upon the function of the pump. Next, since Rv1878 was found not to be essential for the detoxification of Spm, we sought to follow-up on the investigation of the physiological role of Rv1878 along with Rv1877 and Rv0191. Approach: To evaluate the specificity of efflux pumps in the mycobacterial tolerance to Spm, we generated unmarked ∆rv1877 and ∆rv0191 M.tb mutants and evaluated their susceptibility to Spm. To follow up on the investigation of any other physiological roles they may have, we characterized them along with the ∆rv1878 M.tb mutant. Results: The ∆rv1877 mutant was sensitive to Spm stress, while the ∆rv0191 mutant was not. On the other hand, the ∆rv1878 mutant grew better than the wild-type during iron starvation yet was sensitive to cell wall stress. The proteins Rv1877 and Rv1878 seemed to play physiological roles during hypoxia and acidic stress. Lastly, the ∆rv0191 mutant was the only mutant that was sensitive to oxidative stress. Conclusion: The multidrug MFS-type efflux pump Rv1877 is required for Spm detoxification, as opposed to Rv0191 which seems to play a more specific role. Moreover, Rv1878 seems to play a role in the regulation of iron homeostasis and the reconstitution of the cell wall of M.tb. On the other hand, the sensitivity of the ∆rv0191 mutant to oxidative stress, suggests that Rv0191 may be responsible for the transport of low molecular weight thiols. [ABSTRACT FROM AUTHOR]

New drugs are needed to shorten and simplify treatment of tuberculosis caused by Mycobacterium tuberculosis. Metabolic pathways that M. tuberculosis requires for growth or survival during infection represent potential targets for anti-tubercular drug development. Genes and metabolic pathways essential for M. tuberculosis growth in standard laboratory culture conditions have been defined by genome-wide genetic screens. However, whether M. tuberculosis requires these essential genes during infection has not been comprehensively explored because mutant strains cannot be generated using standard methods. Here we show that M. tuberculosis requires the phenylalanine (Phe) and de novo purine and thiamine biosynthetic pathways for mammalian infection. We used a defined collection of M. tuberculosis transposon (Tn) mutants in essential genes, which we generated using a custom nutrient-rich medium, and transposon sequencing (Tn-seq) to identify multiple central metabolic pathways required for fitness in a mouse infection model. We confirmed by individual retesting and complementation that mutations in pheA (Phe biosynthesis) or purF (purine and thiamine biosynthesis) cause death of M. tuberculosis in the absence of nutrient supplementation in vitro and strong attenuation in infected mice. Our findings show that Tn-seq with defined Tn mutant pools can be used to identify M. tuberculosis genes required during mouse lung infection. Our results also demonstrate that M. tuberculosis requires Phe and purine/thiamine biosynthesis for survival in the host, implicating these metabolic pathways as prime targets for the development of new antibiotics to combat tuberculosis. Author summary: Mycobacterium tuberculosis causes more than 10 million new cases of active tuberculosis (TB) disease and ~1.3 million deaths worldwide each year. Individuals with active TB must take a combination of four antibiotics for a minimum of 6–9 months to cure the infection. New anti-tubercular drugs are needed to simplify TB treatment and combat drug resistance. Here, we describe a novel collection of M. tuberculosis mutants lacking metabolic pathways essential for growth in standard laboratory conditions. Using these mutants, a mouse infection model, and deep sequencing we identified those metabolic pathways that M. tuberculosis also requires during infection. We find that M. tuberculosis mutants that cannot synthesize purine nucleotides, riboflavin, or certain amino acids are unable to grow in mice. We also find that mutant strains which cannot synthesize purine nucleotides or the amino acid phenylalanine die rapidly in laboratory cultures without nutrient supplementation, suggesting that new drugs targeting these pathways would kill M. tuberculosis. Overall, our work reveals multiple metabolic pathways that M. tuberculosis requires during infection, which could be pursued as new targets for development of anti-tubercular drugs. [ABSTRACT FROM AUTHOR]

Introduction: Helicobacter pylori (H.pylori, Hp) affects billions of people worldwide. However, the emerging resistance of Hp to antibiotics challenges the effectiveness of current treatments. Investigating the genotypephenotype connection for Hp using next-generation sequencing could enhance our understanding of this resistance. Methods: In this study, we analyzed 52 Hp strains collected from various hospitals. The susceptibility of these strains to five antibiotics was assessed using the agar dilution assay. Whole-genome sequencing was then performed to screen the antimicrobial resistance (AMR) genotypes of these Hp strains. To model the relationship between drug resistance and genotype, we employed univariate statistical tests, unsupervised machine learning, and supervised machine learning techniques, including the development of support vector machine models. Results: Our models for predicting Amoxicillin resistance demonstrated 66% sensitivity and 100% specificity, while those for Clarithromycin resistance showed 100% sensitivity and 100% specificity. These results outperformed the known resistance sites for Amoxicillin (A1834G) and Clarithromycin (A2147), which had sensitivities of 22.2% and 87%, and specificities of 100% and 96%, respectively. Discussion: Our study demonstrates that predictive modeling using supervised learning algorithms with feature selection can yield diagnostic models with higher predictive power compared to models relying on single single-nucleotide polymorphism (SNP) sites. This approach significantly contributes to enhancing the precision and effectiveness of antibiotic treatment strategies for Hp infections. The application of whole-genome sequencing for Hp presents a promising pathway for advancing personalized medicine in this context. [ABSTRACT FROM AUTHOR]

This study assesses the diagnostic efficacy of a Polymerase Chain Reaction (PCR) assay targeting a 1412 bp segment of the 16S rRNA sequence of Brucella melitensis in suspected cases of brucellosis in goats. The research investigates the assay's performance with both serum and whole blood samples in 42 suspected brucellosis cases. Results indicate a significant difference in sensitivity between serum and whole blood samples, with the PCR assay demonstrating higher sensitivity (90%) in serum compared to whole blood (80%). The specificity of the serum PCR assay is determined to be 100%, emphasizing its precision in identifying negative cases. The study strongly recommends the preferential use of serum samples over whole blood for brucellosis diagnosis in goats using the 16S rRNA-based PCR assay, emphasizing the enhanced sensitivity observed in serum samples. These findings contribute to optimizing diagnostic protocols for brucellosis detection in goat populations, with potential implications for veterinary practice and research endeavors. [ABSTRACT FROM AUTHOR]

Super Napier, as a green forage, is becoming very popular among the farming community. A study was undertaken to know the suitability, green fodder production potential of this hybrid in the Kalyana-Karnataka region of Karnataka state. The study was carried out for three continuous years at six different Krishi Vigyan Kendra's of the University of Agricultural Sciences, Raichur. Sixty farmers, ten each in each KVK were purposively selected. Farmers growing Hybrid Napier (DHN-6) and having irrigation facilities were considered for the study. Stem slips of both Co-3 and Super Napier each 5000 per acre were distributed as critical inputs. The parameters namely green fodder yield, palatability, milk yield and its composition in HF cross bred cows and economics were studied. Based on these parameters, it was concluded that Super Napier fodder hybrid is superior, better and more profitable to the farming community compared to Co-3 and Hybrid Napier (DHN-6). [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa (PA) is an important opportunistic pathogen that causes different infections on immunocompromised patients. Within PA accessory genome, differences in virulence, antibiotic resistance and biofilm formation have been described between strains, leading to the emergence of multidrug-resistant strains. The genome sequences of 17 strains isolated from patients with healthcare-associated infections in a Mexican hospital were genomically and phylogenetically analyzed and antibiotic resistance genes, virulence genes, and biofilm formation genes were detected. Fifteen of the 17 strains were resistant to at least two of the carbapenems meropenem, imipenem, and the monobactam aztreonam. The antibiotic resistance (mexA, mexB, and oprM) and the biofilm formation (pslA and pslD) genes were detected in all strains. Differences were found between strains in accessory genome size. The strains had different sequence types, and seven strains had sequence types associated with global high risk epidemic PA clones. All strains were represented in two groups among PA global strains. In the 17 strains, horizontally acquired resistance genes to aminoglycosides and beta-lactams were found, mainly, and between 230 and 240 genes that encode virulence factors. The strains under study were variable in terms of their accessory genome, antibiotic resistance, and virulence genes. With these characteristics, we provide information about the genomic diversity of clinically relevant PA strains. [ABSTRACT FROM AUTHOR]

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Despite the implementation of brucellosis eradication programs in Iran, this disease is still endemic and highly prevalent among ruminants in this country. The infection of small ruminants with Brucella abortus may play a significant role in the continuation of brucellosis among the herds of ruminants. This study investigated the proportion of B. abortus and Brucella melitensis in 150 samples of sheep and goat's raw milk which were obtained from Lorestan and Hamadan provinces in the western part of Iran using the PCR method. The results revealed that among the Brucella spp. positive samples, 26.5% and 73.4% of the samples were infected with B. abortus and B. melitensis, respectively. The incidence rates of B. abortus among the sheep and goats samples were 6.8% and 12.5%, respectively. There was a significant difference between goats and sheep regarding the proportion of B. abortus. Three samples (2%) (2 goats and one sheep) were simultaneously infected with both B. melitensis and B. abortus. This article renews our knowledge about the causative agent of brucellosis in small ruminants and shows that B. abortus has a relatively high prevalence among those animals in the western regions of Iran, and its role as one of the main factors of abortion among small ruminants should not be ignored. The information provided in the present study is important for the surveillance program, as eradication programs and strategies to prevent the spread of B. abortus among small ruminants that have not been vaccinated against this microorganism may be adapted accordingly. [ABSTRACT FROM AUTHOR]

Urbanization is altering landscapes globally at an unprecedented rate. While ecological differences between urban and rural environments often promote phenotypic divergence among populations, it is unclear to what degree these trait differences arise from genetic divergence as opposed to phenotypic plasticity. Furthermore, little is known about how specific landscape elements, such as green corridors, impact genetic divergence in urban environments. We tested the hypotheses that: (1) urbanization, and (2) proximity to an urban green corridor influence genetic divergence in common milkweed (Asclepias syriaca) populations for phenotypic traits. Using seeds from 52 populations along three urban-to-rural subtransects in the Greater Toronto Area, Canada, one of which followed a green corridor, we grew ~ 1000 plants in a common garden setup and measured > 20 ecologically-important traits associated with plant defense/damage, reproduction, and growth over four years. We found significant heritable variation for nine traits within common milkweed populations and weak phenotypic divergence among populations. However, neither urbanization nor an urban green corridor influenced genetic divergence in individual traits or multivariate phenotype. These findings contrast with the expanding literature demonstrating that urbanization promotes rapid evolutionary change and offer preliminary insights into the eco-evolutionary role of green corridors in urban environments. [ABSTRACT FROM AUTHOR]

Objective adherence data, to motivate people with asthma to adhere to their medication regimen.Methods: A participatory user-centered design approach was employed, involving end-users and other stakeholders throughout the study. The approach consisted of four phases. Semi-structured interviews and a survey were conducted to understand user needs and reasons for poor adherence (Phase 1: define). Key themes were identified, leading to the formulation of behavior change strategies and design and game requirements. Several design directions were ideated, resulting in a concept for a serious game (Phase 2: ideate). Two rounds of user-tests were performed to evaluate a prototype of the serious game in terms of usability, perceived impact on medication adherence and motivation (Phase 3: prototype and Phase 4: evaluate).Results: Findings from semi-structured interviews (n = 6) and the online survey (n = 20) revealed that people's non-adherence was often attributed to the perception of asthma as an episodic condition, the delayed experienced effect of maintenance inhalers, and lack of knowledge regarding difference of effect between maintenance and reliever inhalers. The study used behavior change strategies to translate these insights into design requirements for the development of the narrative-based persuasive game Ademgenoot. This six-week challenge-based game combines various behavior change strategies, including personal goal setting and continuous visual feedback, as well as persuasive game design elements, such as a narrative and rewards, with the aim of enhancing motivation to adhere to their medication regimen. User-testing (n = 8; round 1 and 2) showed that Ademgenoot is feasible in clinical practice and has the potential to support people with mild asthma in adherence to their maintenance medication.Discussion: Future efforts should be directed towards a larger evaluation to assess the impact on motivation and inhaler use behaviour.Plain Language Summary: The goal of this study was to create a serious game that encourages people with asthma to take their medication regularly. During the study, we worked closely with individuals who have asthma and other stakeholders throughout the study. We conducted interviews and surveys to understand why people have difficulties using their maintenance inhaler as prescribed by their doctor. Based on the feedback we received, we developed a serious game called "Ademgenoot". The game uses information on inhaler use automatically collected with a device attached to the inhaler. The game includes features like personal goals and visual feedback on inhaler use to motivate users to take their medication consistently. We tested a prototype of the game with users to see if it was easy to use and if it motivated them to use their maintenance inhaler. The results showed that Ademgenoot is a viable option for helping individuals with mild asthma stay on track with their medication.Key words: medication adherence, serious game design, behavior change, adherence monitoring, participatory design [ABSTRACT FROM AUTHOR]

Despite the great diversity of malonate semialdehyde decarboxylases (MSADs), one of five subgroups of the tautomerase superfamily (TSF) found throughout the biosphere, their distribution among strains within the genus Mycobacterium remains unknown. In this study, we sought to investigate the phylogenetic distribution of MSAD genes of mycobacterial species via genome analysis of 192 different reference Mycobacterium species or subspecies retrieved from NCBI databases. We found that in a total of 87 of 192 strains (45.3%), MSAD-1 and MSAD-2 were distributed in an exclusive manner among Mycobacterium species except for 12 strains, including Mycobacterium chelonae members, with both in their genome. Of note, Mycobacterium strains better adapted to the host and of high virulence potential, such as the Mycobacterium tuberculosis complex, Mycobacterium leprae, Mycobacterium marinum, Mycobacterium ulcerans, and Mycobacterium avium subsp. paratuberculosis, had no orthologs of MSAD in their genome, suggesting MSAD loss during species differentiation in pathogenic slow growing Mycobacterium. To investigate the MSAD distribution among strains of M. avium subspecies, the genome sequences of a total of 255 reference strains from the four subspecies of M. avium (43 of subspecies avium, 162 of subspecies hominissuis, 49 of subspecies paratuberculosis, and 1 of subspecies silvaticum) were further analyzed. We found that only 121 of 255 strains (47.4%) had MSADs in their genome, with none of the 49 M. avium subsp. paratuberculosis strains having MSAD genes. Even in 13 of 121 M. avium strains with the MSAD-1 gene in their genome, deletion mutations in the 98th codon causing premature termination of MSAD were found, further highlighting the occurrence of MSAD pseudogenization during species or subspecies differentiation of M. avium. In conclusion, our data indicated that there are two distinct types of MSADs, MSAD1 and MSAD-2, among strains in the Mycobacterium genus, but more than half of the strains, including pathogenic mycobacteria, M. tuberculosis and M. leprae, have no orthologs in their genome, suggesting MSAD loss during host adaptation of pathogenic mycobacteria. In the future, the role of two distinct MSADs, MSAD-1 and MSAD-2, in mycobacterial pathogenesis or evolution should be investigated. [ABSTRACT FROM AUTHOR]

Background: Tuberculosis may reoccur due to reinfection or relapse after initially successful treatment. Distinguishing the cause of TB recurrence is crucial to guide TB control and treatment. This study aimed to investigate the source of TB recurrence and risk factors related to relapse in Hunan province, a high TB burden region in southern China. Methods: A population-based retrospective study was conducted on all culture-positive TB cases in Hunan province, China from 2013 to 2020. Phenotypic drug susceptibility testing and whole-genome sequencing were used to detect drug resistance and distinguish between relapse and reinfection. Pearson chi-square test and Fisher exact test were applied to compare differences in categorical variables between relapse and reinfection. The Kaplan--Meier curve was generated in R studio (4.0.4) to describe and compare the time to recurrence between different groups. p < 0.05 was considered statistically significant. Results: Of 36 recurrent events, 27 (75.0%, 27/36) paired isolates were caused by relapse, and reinfection accounted for 25.0% (9/36) of recurrent cases. No significant difference in characteristics was observed between relapse and reinfection (all p >0.05). In addition, TB relapse occurs earlier in patients of Tu ethnicity compared to patients of Han ethnicity (p <0.0001), whereas no significant differences in the time interval to relapse were noted in other groups. Moreover, 83.3% (30/36) of TB recurrence occurred within 3 years. Overall, these recurrent TB isolates were predominantly pan-susceptible strains (71.0%, 49/69), followed by DR-TB (17.4%, 12/69) and MDR-TB (11.6%, 8/69), with mutations mainly in codon 450 of the rpoB gene and codon 315 of the katG gene. 11.1% (3/27) of relapse cases had acquired new resistance during treatment, with fluoroquinolone resistance occurring most frequently (7.4%, 2/27), both with mutations in codon 94 of gyrA. Conclusion: Endogenous relapse is the main mechanism leading to TB recurrences in Hunan province. Given that TB recurrences can occur more than 4 years after treatment completion, it is necessary to extend the post-treatment follow-up period to achieve better management of TB patients. Moreover, the relatively high frequency of fluoroquinolone resistance in the second episode of relapse suggests that fluoroquinolones should be used with caution when treating TB cases with relapse, preferably guided by DST results. [ABSTRACT FROM AUTHOR]

Background: Brucellosis is one of the major zoonotic problems that exist worldwide. Brucellosis is clinically characterized by metritis, mastitis, repeat breeding, abortion in the last trimester of pregnancy, retention of placenta and reduced milk production in the female whereas epididymitis, orchitis and sterility in male. In humans can be highly variable, ranging from nonspecific, flu-like symptoms to undulant fever, arthritis, orchitis and epididymitis. Methods: A total of 567 bovine serum samples was taken from four districts of Brij region of UP. All the samples were processed to detection of prevalence of brucellosis by RBPT, STAT ELISA and confirmation of genes bcsp31, 16SrRNA, omp2 and IS711 by PCR. Result: The prevalence of brucellosis was found to be 07.93% (31/391), 08.69% (34/391) and 10.74% (42/391) shows positive by RBPT, STAT and I- ELISA respectively. In buffalo Out of 176 tested serum sample the seroprevalence was found to be 09.66% (17/176), 10.79% (19/176) and 12.5% (22/176) positive by RBPT, STAT and I- ELISA respectively. Out of 567 samples 18 were positive for Brucella genus specific gene. The higher prevalence of the disease in this region increases the risk of zoonotic transmission and it implies a serious threat to the human population as well as the huge impact on economy due to loss of productivity as well as loss of livestock population. [ABSTRACT FROM AUTHOR]

Metabolic dysregulation in Mycobacterium tuberculosis results in increased macrophage apoptosis or pyroptosis. However, mechanistic links between Mycobacterium virulence and bacterial metabolic plasticity remain ill defined. In this study, we screened random transposon insertions of M. bovis BCG to identify mutants that induce pyroptotic death of the infected macrophage. Analysis of the transposon insertion sites identified a panel of fdr (functioning death repressor) genes, which were shown in some cases to encode functions central to Mycobacterium metabolism. In-depth studies of one fdr gene, fdr8 (BCG3787/Rv3727), demonstrated its important role in the maintenance of M. tuberculosis and M. bovis BCG redox balance in reductive stress conditions in the host. Our studies expand the subset of known Mycobacterium genes linking bacterial metabolic plasticity to virulence and also reveal that the broad induction of pyroptosis by an intracellular bacterial pathogen is linked to enhanced cellular immunity in vivo. [ABSTRACT FROM AUTHOR]

Animal reproduction biotechniques are important tools for the technological advancement of livestock, as they allow the selection of the reproductive potential of superior quality females and males; however, infectious diseases that have a predilection for the reproductive system can be a hindrance for the use of these technologies. Therefore, the present study aimed to detect Brucella spp. in the ovarian follicular fluid of brucellosis-positive bovine cows. A total of 47 bovine ovarian follicular fluid aspirates from cows, positive in tests for brucellosis and from Brucella-positive herd, were submitted to PCR. The primers used in the PCR were specific to the genus Brucella (bcsp31 gene). All 47 bovine aspirates were negative for Brucella spp. 0.00% (95% CI: 0.00-4.00%). Our results demonstrated that Brucella spp. was absent in the ovarian follicular fluid from seropositive cows, which indicates that Brucella spp.-infected cows could be used for reproductive biotechnologies carried out with follicular aspirates. Future studies are needed to more precisely evaluate the feasibility and safety of using these oocytes from brucellosis-seropositive cows to transfer embryos to heifers/cows not infected by Brucella, aiming to produce calves free of the infection., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

Background: Immigration is considered as a risk factor of tuberculosis (TB). Qom province receives millions of pilgrims and significant numbers of immigrants each year. Most of the immigrants to Qom, arrive from neighboring TB-endemic countries. This study aimed to identify the current circulating Mycobacterium tuberculosis genotypes in Qom province using 24-locus MIRU-VNTR genotyping. Methods: Eighty six M. tuberculosis isolates were collected during 2018–2022 from patients referring to Qom TB reference laboratory. The DNA of isolates was extracted and followed by 24 loci MIRU-VNTR genotyping which performed using the web tools available on MIRU-VNTRplus. Results: Of 86 isolates, 39 (45.3%) were of Delhi/CAS genotype, 24 (27.9%) of NEW-1, 6 (7%) of LAM, 6 (7%) of Beijing, 2 (2.3%) of UgandaII, 2 (2.3%) of EAI, 1 of S (1.2%) and 6 (7%) did not match with profiles present in MIRUVNTRplus database. Conclusions: About half of the isolates belong to Afghan immigrants; which warns health policy makers about the future situation of TB in Qom. Also, the similarity of Afghan and Iranian genotypes provides evidence that immigrants partake in the circulation of M. tuberculosis. This study underpin the studies about the circulating M. tuberculosis genotypes, their geographical distribution, the association of TB risk factors with these genotypes and the impact of immigration on the situation of TB in Qom province. [ABSTRACT FROM AUTHOR]

Introduction: M. aviumsubsp. hominissuis (M. avium) is an intracellular, facultative bacterium known to colonize and infect the human host through ingestion or respiratory inhalation. The majority of pulmonary infections occur in association with pre-existing lung diseases, such as bronchiectasis, cystic fibrosis, or chronic obstructive pulmonary disease. M. avium is also acquired by the gastrointestinal route in immunocompromised individuals such as human immunodeficiency virus HIV-1 patients leading to disseminated disease. A hallmark of M. avium pulmonary infections is the ability of pathogen to form biofilms. In addition, M. avium can reside within granulomas of low oxygen and limited nutrient conditions while establishing a persistent niche through metabolic adaptations. Methods: Bacterial metabolic pathways used by M. avium within the host environment, however, are poorly understood. In this study, we analyzed M. avium proteome with a focus on core metabolic pathways expressed in the anaerobic, biofilm and aerobic conditions and that can be used by the pathogen to transition from one environment to another. Results: Overall, 3,715 common proteins were identified between all studied conditions and proteins with increased synthesis over the of the level of expression in aerobic condition were selected for analysis of in specific metabolic pathways. The data obtained from the M. avium proteome of biofilm phenotype demonstrates in enrichment of metabolic pathways involved in the fatty acid metabolism and biosynthesis of aromatic amino acid and cofactors. Here, we also highlight the importance of chloroalkene degradation pathway and anaerobic fermentationthat enhance during the transition of M. avium from aerobic to anaerobic condition. It was also found that the production of fumarate and succinate by MAV_0927, a conserved hypothetical protein, is essential for M. avium survival and for withstanding the stress condition in biofilm. In addition, the participation of regulatory genes/proteins such as the TetR family MAV_5151 appear to be necessary for M. avium survival under biofilm and anaerobic conditions. Conclusion: Collectively, our data reveal important core metabolic pathways that M. avium utilize under different stress conditions that allow the pathogen to survive in diverse host environments. [ABSTRACT FROM AUTHOR]

Tuberculosis is a major global threat to human health. Since the widely used BCG vaccine is poorly effective in adults, there is a demand for the development of a new type of boost tuberculosis vaccine. We designed a novel intranasal tuberculosis vaccine candidate, TB/FLU-04L, which is based on an attenuated influenza A virus vector encoding two mycobacterium antigens, Ag85A and ESAT-6. As tuberculosis is an airborne disease, the ability to induce mucosal immunity is one of the potential advantages of influenza vectors. Sequences of ESAT-6 and Ag85A antigens were inserted into the NS1 open reading frame of the influenza A virus to replace the deleted carboxyl part of the NS1 protein. The vector expressing chimeric NS1 protein appeared to be genetically stable and replication-deficient in mice and non-human primates. Intranasal immunization of C57BL/6 mice or cynomolgus macaques with the TB/FLU-04L vaccine candidate induced Mtb-specific Th1 immune response. Single TB/FLU-04L immunization in mice showed commensurate levels of protection in comparison to BCG and significantly increased the protective effect of BCG when applied in a "prime-boost" scheme. Our findings show that intranasal immunization with the TB/FLU-04L vaccine, which carries two mycobacterium antigens, is safe, and induces a protective immune response against virulent M. tuberculosis. [ABSTRACT FROM AUTHOR]

Background and Objective. Tuberculosis (TB) is a major threat to human health, especially in developing countries. Its susceptibility and progression depend on interactions between mycobacterium tuberculosis, host immune system, and genetic and environmental factors. Up to now, many studies have presented the association between TB susceptibility and host genetic polymorphisms, but never regarding CD59 gene, which is an essential complement regulator. This study investigated the relationship between multiple CD59 single nucleotide polymorphisms (SNPs) and susceptibility to TB among Chinese patients. Methods. A case–control study was conducted to investigate the SNPs at CD59 rs1047581, rs7046, rs2231460, rs184251026, rs41275164, rs831633, rs704700, rs41275166, and rs10768024 by sequence-specific primer-polymerase chain reaction (SSP-PCR) in 900 tuberculosis patients and 1,534 controls. Results. The minor allele frequencies at rs2231460, rs184251026, rs41275164, and rs41275166 were extremely low both in the Cases (0.00%–0.61%) and in the Controls (0.07%–0.43%), comparatively at rs1047581, rs7046, rs831633, rs704700, and rs10768024 were notably higher both in the Cases (8.23%–48.39%) and in the Controls (8.57%–47.16%). Among the nine SNPs, only homozygous CC genotype at rs10768024 showed a significant protective effect against TB than homozygous TT genotype (OR(95% CI) = 0.59(0.38, 0.91), χ2 = 5.779, P = 0.016), and homozygous TT and heterozygous CT genotypes showed a significant risk of TB infection in the recessive model (OR(95% CI) = 1.68(1.10, 2.56), χ2 = 5.769, P = 0.016). Further analysis verified that rs10768024 CC genotype independently related to TB susceptibility (OR(95% CI) = 0.60(0.39, 0.91), Wald χ2 = 5.664, P = 0.017) in multivariate logistic regression analysis, and its genetic mutation was independent of the other SNPs (r2 = 0.00–0.20) in haplotype analysis. Conclusions. The first investigation of the CD59 gene and susceptibility to TB suggests a significant risk with homozygous TT and heterozygous CT genotypes at rs10768024 loci. The homozygous CC mutation at rs10768024 loci showed a significant protection against TB susceptibility. [ABSTRACT FROM AUTHOR]

The demand for eye-safe 1550 nm pulsed semiconductor laser-emission modules is increasing in the field of active laser detection, owing to their long range and high precision. The high power and narrow pulse of these modules can significantly improve the distance and accuracy of active-laser detection. Here, we propose an equivalent circuit model of a multiquantum-well laser based on the structure of a laser device. We developed a design method for 1550 nm pulsed semiconductor laser-emission modules according to the equivalent circuit model of an InGaAlAs laser. In this method, the module design was divided into laser chip and laser-driver levels for optimization and simulation. At the chip level, a high-output power laser chip with optimal cavity length and optical facet coating coefficients was obtained. At the laser-driver level, the model was applied to a drive circuit to provide direct narrow optical pulses. Finally, a laser-emission module was fabricated based on the optimal design results. In addition to the power-current features of the actual laser, the critical voltage of the emission module and laser pulses were tested. By comparing the test and simulation results, the effectiveness of the proposed method was confirmed. [ABSTRACT FROM AUTHOR]

Introduction: Diabetes is a potent risk factor for the activation of latent tuberculosis and worsens the tuberculosis (TB) treatment outcome. The major reason for mortality and morbidity in diabetic patients is due to their increased susceptibility to TB. Thus, the study was conducted to understand the crosstalk between M. tuberculosis and its host upon latent tuberculosis infection and under hyperglycemic conditions or diabetes. Methods: An animal model was employed to study the relationship between latent tuberculosis and diabetes. BCG immunization was done in mice before infection with M. tuberculosis, and latency was confirmed by bacillary load, histopathological changes in the lungs and gene expression of hspX, tgs1, tgs3 and tgs5. Diabetes was then induced by a single high dose of streptozotocin (150 mg/kg body weight). Host factors, like various cytokines and MMPs (Matrix metalloproteinases), which play an important role in the containment of mycobacterial infection were studied in vivo and in vitro. Results: A murine model of latent TB was developed, which was confirmed by CFU counts (<104 in the lungs and spleen) and granuloma formation in lungs in the latent TB group. Also, the gene expression of hspX, tgs1, and tgs5 was upregulated, and after diabetes induction, blood glucose levels were >200 mg/dl. An in vitro study employing a THP-1 macrophage model of latent and active tuberculosis under normal and high glucose conditions showed that dormant bacilli were better contained in the presence of 5.5 mM glucose concentration as compared with active bacilli. However, the killing and restriction efficiency of macrophages decreased, and CFU counts increased significantly with an increase in glucose concentration. Discussion: The decreased levels of MCP-1, decreased expression of mmp-9, and increased expression of mmp-1 in the latent group at high glucose concentrations could explain the failure of granuloma formation at high glucose conditions. [ABSTRACT FROM AUTHOR]

The facultative intracellular pathogen Brucella and the free-living bacteria Ochrobactrum are both α-proteobacteria and very close to each other. A group of researchers recently clustered Ochrobactrum strains into the genus Brucella according to a BLAST distance approach. Thus, we performed a deeper comparative genetic analysis for eleven Ochrobactrum strains and twelve different Brucella isolates to demonstrate important differences between these bacteria. In addition to the clear differences between Brucella and Ochrobactrum, like the differences in genes contents, and different genome sizes, the Brucella-specific gene bscp31 was not found in Ochrobactrum, as well as other important Brucella-specific proteins and virulence factors. Differences in antimicrobial resistance genes content and the presence or absence of plasmids were obvious between Brucella and Ochrobactrum spp. Genome alignment of Brucella spp. and Ochrobactrum spp. revealed a genome similarity of 85.7% maximum, whereas all analyzed Brucella spp. in this study had a similarity of 97.6-99.9%, and all compared Ochrobactrum spp. 82.6-98.0%. Because of these facts mentioned in this work, Brucella and Ochrobactrum should be considered separate genera. [ABSTRACT FROM AUTHOR]

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is a tenacious pathogen that has latently infected one third of the world's population. However, conventional TB treatment regimens are no longer sufficient to tackle the growing threat of drug resistance, stimulating the development of innovative anti-tuberculosis agents, with special emphasis on new protein targets. The Mtb genome encodes ~4000 predicted proteins, among which many enzymes participate in various cellular metabolisms. For example, more than 200 proteins are involved in fatty acid biosynthesis, which assists in the construction of the cell envelope, and is closely related to the pathogenesis and resistance of mycobacteria. Here we review several essential enzymes responsible for fatty acid and nucleotide biosynthesis, cellular metabolism of lipids or amino acids, energy utilization, and metal uptake. These include InhA, MmpL3, MmaA4, PcaA, CmaA1, CmaA2, isocitrate lyases (ICLs), pantothenate synthase (PS), Lysine-ε amino transferase (LAT), LeuD, IdeR, KatG, Rv1098c, and PyrG. In addition, we summarize the role of the transcriptional regulator PhoP which may regulate the expression of more than 110 genes, and the essential biosynthesis enzyme glutamine synthetase (GlnA1). All these enzymes are either validated drug targets or promising target candidates, with drugs targeting ICLs and LAT expected to solve the problem of persistent TB infection. To better understand how anti-tuberculosis drugs act on these proteins, their structures and the structure-based drug/inhibitor designs are discussed. Overall, this investigation should provide guidance and support for current and future pharmaceutical development efforts against mycobacterial pathogenesis. [ABSTRACT FROM AUTHOR]

The specific features of the reactivation of "non-culturable" (NC) Mycobacterium tuberculosis (MTB) cells for the following multiplication in liquid medium were defined, as well as the approach for the quantification of these cells by culture methods was suggested. When NC cells obtained in vitro were inoculated into the standard Sauton's liquid medium, a prolonged (up to 20-day) lag phase was observed, in which no cell multiplication was occurred. During the lag phase, dormant cells secreted substances of an unknown nature that inhibited or completely arrested the reactivation and growth of cultures when the initial dormant cell concentration was above 107–108 cells/mL. Addition of meat-peptone broth (3.25 g/L) into a standard Sauton's medium with a 10-fold reduced concentration of glycerol significantly stimulated the reactivation and multiplication of the initially dormant cells inoculated at a concentration above the threshold. This modification of the medium made it possible to detect up to 103 times more cells in the MTB population by the method of the Most Probable Number (MPN) of cells. An increased number of the detected dormant cells in the population (up to an average of 2.5 orders of magnitude) was also achieved by adding RpfB recombinant protein (5 ng/mL) (a protein of the Rpf family, resuscitation promoting factor of dormant bacteria) to the reactivation medium. Perhaps, the action of the Rpf enzyme is related to the products of its enzymatic activity, since an increased MPN value in the dormant cells population was also observed when the products of mycobacteria peptidoglycan hydrolysis obtained by the coaction of RpfB and endopeptidase RipA were added. The addition of sonicated peptidoglycan fragments at a concentration up to 1 μg/mL had a similar effect. These results may be used in approaches to optimize the liquid media composition and culture conditions to identify the pathogen of tuberculosis in clinical samples in a nonculturable state. [ABSTRACT FROM AUTHOR]

This study examines three noncompliant cases occurred in Wuhan, China during the outbreak of the COVID-19 in 2020, including the Li Wenliang (李文亮) case, the Chen Qiushi (陈秋实) case, and the Huang Dengying (黄登英) case. These cases, albeit with different natures, broke out during the early stage of Wuhan ™s epidemic and posted threat and challenges to the Chinese authorities to different degrees. Based on open source information, this study focuses on the issues of trust and examines trust at three different levels (interpersonal trust, public trust, and political trust) among interactions by three groups of actors, including individuals involved in each case, the general public (e.g. how the public perceived and evaluated each case), and the authorities (e.g. how governments at the local and central levels perceived and handled each case). Two research questions are addressed: 1) How did trust at different levels play out in each case? 2) How did Chinese authorities respond to and handle threat and public trust challenges? Policy implications are drawn. [ABSTRACT FROM AUTHOR]

Objective  We evaluated the usefulness of a three-dimensional (3D) interactive atlas to illustrate and teach surgical skull base anatomy in a clinical setting. Study Design  A highly detailed atlas of the adult human skull base was created from multiple high-resolution magnetic resonance imaging (MRI) and computed tomography (CT) scans of a healthy Caucasian male. It includes the parcellated and labeled bony skull base, intra- and extracranial vasculature, cranial nerves, cerebrum, cerebellum, and brainstem. We are reporting retrospectively on our experiences with employing the atlas for the simulation and teaching of neurosurgical approaches and concepts in a clinical setting. Setting  The study was conducted at the University Hospital Mainz, Germany, and Hirslanden Hospital, Zürich, Switzerland. Participants  Medical students and neurosurgical residents participated in this study. Results  Handling the layered graphical user interface of the atlas requires some training; however, navigating the detailed 3D content from intraoperative perspectives led to quick comprehension of anatomical relationships that are otherwise difficult to perceive. Students and residents appreciated the collaborative learning effect when working with the atlas on large projected screens and markedly improved their anatomical knowledge after interacting with the software. Conclusion  The skull base atlas provides an effective way to study essential surgical anatomy and to teach operative strategies in this complex region. Interactive 3D computer graphical environments are highly suitable for conveying complex anatomy and to train and review surgical concepts. They remain underutilized in clinical practice. [ABSTRACT FROM AUTHOR]