Mycobacteria

BACKGROUND AND PURPOSE:
Phagosomal maturation arrest is known to play a central role in the survival of pathogenic mycobacteria within macrophages. The maturation arrest of mycobacterial phagosome results from the retention of tryptophan-aspartate-containing coat protein (TACO) on this organelle, enabling successful replication of the pathogen. We have shown earlier that vitamin D(3) and retinoic acid (RA) down-regulate TACO gene transcription in a dose-dependent manner.
METHODS:
In this study, we analyzed the promoter region of TACO gene using bioinformatics tools and observed that the vitamin D receptor (VDR)/retinoid-X-receptor (RXR) response sequence was highly functional. We also evaluated the effect of treatment with vitamin D(3)/RA on Mycobacterium tuberculosis entry and survival in cultured human macrophages.
RESULTS:
TACO gene down-regulation observed with vitamin D(3)/RA treatment occurred through modulation of this gene via the VDR/RXR response sequence present in the promoter region of TACO gene. Treatment of macrophages with vitamin D(3)/RA allows maturation of mycobacterial phagosome, leading to degradation of the pathogen.
CONCLUSIONS:
Our results elucidate the mechanism of TACO gene down-regulation observed with vitamin D(3)/RA. Furthermore, the results revealed that vitamin D(3)/RA treatment inhibits mycobacterial entry as well as survival within macrophages, possibly through rescue of phagosome maturation arrest. The developing knowledge in this area suggests that vitamin D(3)/RA may be of importance in the treatment of intracellular infection, particularly tuberculosis.

The skin of amphibians has unique structural properties and physiologic functions that make amphibians particularly sensitive to environmental perturbations and cutaneous injury. This inherent sensitivity makes the skin a critically important site of evaluation in both clinical and postmortem examination of the amphibian patient. Many of the described amphibian skin diseases can be traced back to factors related to captive husbandry. Cutaneous injuries, alterations in water quality (such as pH, ammonia), and exposure to chemical irritants, are all potential hazards of the captive environment that can lead to skin irritation. Furthermore, many infectious diseases of amphibian skin such as "red leg" syndrome, mycobacteriosis, and saprolegniasis are often secondary to environmental factors and require correction of underlying lapses in husbandry for control. Other amphibian skin diseases, such as chytridiomycosis and systemic iridovirus infections are not yet associated with specific environmental cofactors but have been associated with significant mortalities in captive and/or wild amphibians. Amphibian skin diseases often have a similar gross appearance with cutaneous hyperemia and discoloration, cutaneous papules and nodules, and ulceration being among the most common presentations. This review emphasizes conditions most often encountered in amphibian dermatology as well as recently recognized diseases such as chytridiomycosis.

IN PRESS

Every seventy-two seconds someone in America develops Alzheimer’s disease (AD). And it has been said that almost everyone living long enough will eventually show evidence of Alzheimer’s disease. Thus far its cause has remained elusive.
Nevertheless, recently, study after study, in which scientists have injected human Alzheimer diseased brain tissue into mice and other laboratory animals that later developed the disease have left little doubt that Alzheimer’s arises from an infectious process, the focus of debate seeming to be which particular disease. But clearly, it must be a disease that is not only statistically widespread in the world today, but that was also prevalent at the time of Dr. Alois Alzheimer.
To be sure, it was German neuropathologist Oskar Fischer of the Prague school of neuropathology, Alzheimer’s great rival, who was the first to suggest that infection might be causative for Alzheimer’s. Fischer’s credentials: he was the co-discoverer of Alzheimer’s disease and tirelessly did autopsies on the brains of patients with Alzheimer’s.
Fischer’s infectious view never gained immediate popularity, although today, more than a century later, a volume of data supporting such an approach has begun to accumulate. But was Fischer’s specific microbe on the right track to discovering the cause of Alzheimer’s to begin with? The evidence uncovered in this book seems to suggest that he was considerably closer than anyone else—either then or since.
In this volume, doctor and medical researcher Lawrence Broxmeyer presents a stunning hypothesis about the bacterial nature of Alzheimer’s disease, forgotten long ago. Immensely readable and enormously well-researched, this work of medical history should change the way we think about, diagnose, and treat this debilitating condition.

KIRKUS REVIEWS: "The author is an internist and a medical researcher, and his double mastery of both the scientific minutiae and historical nuances of his subject matter is breathtaking. This is more than an account of a scientific debate--it's also an examination of the sometimes--unempirical way that such debate proceeds, as it's conducted by human beings with agendas of their own. Although it's a relatively short book--less than 200 pages of text--it is by no means a quick read. Nevertheless, readers with strong science backgrounds will be impressed by the author's undeniable competence, as well as his journalistic approach to chartering the evolution of thought regarding one of our era's most challenging diseases."

The TSE'S or transmissible spongiform enchephalopathies, include bovine spongiform encephalopathy (also called BSE or " mad cow disease "), Creutzfeldt– Jakob disease (CJD) in humans, and " scrapie " in sheep or goats (caprine spongiform encephalopathy). They remain a mystery, their cause still hotly debated. Current mad cow diagnosis lies solely in the detection of late appearing " prions " , an acronym for hypothesized, geneless, misfolded proteins, somehow claimed to cause the disease. Yet laboratory preparations of prions contain other things, which could include unidentified bacteria or viruses. And the only real evidence that prion originator Stanley Prusiner had in his original paper that the disease agent behind " Scrapie " in sheep and goats was devoid of DNA or RNA– was based upon the fact that he couldn't find any. Furthermore, the rigors of prion purification alone, might, in and of themselves, have killed any causative microorganism and Heino Dringer, who did pioneer work on their nature, candidly predicts " it will turn out that the prion concept is wrong. " Roels and Walravens as well as Hartly traced Mad Cow to Mycobacterium bovis. Moreover, epidemiologic maps of the origins and peak incidence of Mad Cow in the UK, suggestively match those of England's areas of highest bovine tuberculosis, the Southwest. The neurotaxic potential of bovine tuberculosis has for some time been well known. By 1911 Alois Alzheimer called attention to " a characteristic condition of the cortical tissue which Fischer referred to as 'spongy cortical wasting " in Alzheimer's disease (AD). But behind AD, Fischer suspected a microbe called Streptothrix which was constantly being mistaken and confused for tuberculosis. Our present investigation of the TSEs clearly shows cell-wall-deficient (CWD) tubercular mycobacteria present, verified by molecular analysis, ELISA, PCR and microscopy to cause spongiform encephalopathy.

Bovine tuberculosis (BTB) is endemic in Ethiopian livestock. Disease prevalence varies greatly by species, geography, husband-ry system and diagnostic methods used. A substantial body of work has been done on BTB prevalence in Ethiopia over the last decade, the true burden of BTB however, is still largely unknown. This review highlighted gaps and weakness in BTB diagnostics and the urgent need to standardize/harmonize testing methods. It discusses the prevalence of M. tuberculosis and Non Tuber-culous Mycobacteria (NTM) in livestock and their possible interference with the BTB diagnostic methods. This paper finally discusses possible control options for the disease, particularly in the intensive milk farm systems in the wake of increased and or emerging dairy farms in the country.

Alois Alzheimer might have mentioned plaques and tangles in a single short paper on pre-senile dementia in 1907, but it was the co-discover of Alzheimer's disease (AD), Oskar Fischer, who in that same year far more extensively reported neuritic plaque in 12 cases of senile dementia, a condition which he and many others refused to differentiate from Alzheimer's " pre-senile " dementia. Fischer, Alzheimer's great rival, speculated that for the most part these plaques, found only in senile demented patients, caused their dementia. Moreover, Fischer felt such cerebral plaque to be the result of an infection and was very specific as to the sort of infection that might be involved. He felt that he had spotted, throughout his brain autopsies, a tubercular-like Actinobacteria then called Streptothrix (Actinomycosis), often and repeatedly confused with the filamentous cell-wall-deficient forms of the tubercular bacilli. At this point for Fischer, this was the possible infectious cause of AD. To be sure, Oskar Fischer was the first on record to suggest that chronic infection might be causative for what we today call AD. Fischer's infectious view never gained immediate popularity, although today, more than a century later, a volume of data supporting such an approach has begun to accumulate. But was Fischer's specific microbe on the right track to discovering the cause of AD to begin with? Documents uncovered since then seem to suggest that he was considerably closer than anyone else—either then or since. In June of 2017, a University of Bristol study in the UK found a 5 to 10-fold increase in Actinobacteria (order Actinomycetales) population in postmortem AD brains compared with controls [1]. Oskar Fischer's Steptothrix was an Actinobacteria as well, as was the filamentous tuberculosis that Streptothrix was so often mistaken for. But specifically, the Bristol group found an Actinobactor called Propionibacterium acnes (P. Acnes) in autopsied AD brains—a common organism, traditional felt to be non-pathogenic and commonly found on our skin which can

Background: Tuberculosis (TB) is preventable and completely curable major public health problem in the world. Even though the number of TB death decreased by 22% in the past fifteen years it remains one of the top 10 causes of death and the leading cause from a single infectious agent, ranking above HIV/AIDS. It is the largest killer among communicable diseases in age group of 15 to 49years, when humans are most productive. Ethiopia is among the 22 high TB-burden and among the 27 high MDR-TB burden countries in the world with case detection and treatment outcome vary across regions.

Tuberculosis (TB) caused by Mycobacterium bovis is a re-emerging disease of livestock that is of major economic importance worldwide, as well as being a zoonotic risk. There is significant heritability for host resistance to bovine TB (bTB) in dairy cattle. To identify resistance loci for bTB, we undertook a genome-wide association study in female Holstein–Friesian cattle with 592 cases and 559 age-matched controls from case herds. Cases and controls were categorised into distinct phenotypes: skin test and lesion positive vs skin test negative on multiple occasions, respectively. These animals were genotyped with the Illumina BovineHD 700K BeadChip. Genome-wide rapid association using linear and logistic mixed models and regression (GRAMMAR), regional heritability mapping (RHM) and haplotype-sharing analysis identified two novel resistance loci that attained chromosome-wise significance, protein tyrosine phosphatase receptor T (PTPRT; P=4.8 × 10−7) and myosin IIIB (MYO3B; P=5.4 × 10−6). We estimated that 21% of the phenotypic variance in TB resistance could be explained by all of the informative single-nucleotide polymorphisms, of which the region encompassing the PTPRT gene accounted for 6.2% of the variance and a further 3.6% was associated with a putative copy number variant in MYO3B. The results from this study add to our understanding of variation in host control of infection and suggest that genetic marker-based selection for resistance to bTB has the potential to make a significant contribution to bTB control.

Sözcükte anlam aslında anlam bilgisine dahil olup Kpss'de genelde sözcüğün cümle içinde kazandığı anlam bakımından sorular sorulmaktadır. Dilimizde bir sözcüğün tek başına genelde bir anlamı yoktur. Bu sözcükler ya da söz öbekleri cümle içinde anlam kazanmaktadır ve Kpss sınavında da bu anlam üzerinde durulmaktadır. Kpss sınavında bu tarz soruların sorulma sebebi deaslında kişinin ezberini değil anlama yeteneğini ölçmektir.

Rapidly growing mycobacteria (RGM) are common soil saprophytes, but certain strains cause infections in human and animals. The
infections due to RGM have been increasing in past decades and are often difficult to treat. The susceptibility to antibiotics is regularly
evaluated in clinical isolates of RGM, but the data on soil RGM are missing. The objectives of this study was to isolate RGM from four
grassland soils with different impact of manuring, and assess their resistance to antibiotics and the ability to grow at 37°C and 42°C.
Since isolation of RGM from soil is a challenge, a conventional decontamination method (NaOH/malachite green/cycloheximide) and a
recent method based on olive oil/SDS demulsification were compared. The olive oil/SDS method was less efficient, mainly because of
the emulsion instability and plate overgrowing with other bacteria. Altogether, 44 isolates were obtained and 23 representatives of
different RGM genotypes were screened. The number of isolates per soil decreased with increasing soil pH, consistently with previous
findings that mycobacteria were more abundant in low pH soils. Most of the isolates belonged to the Mycobacterium fortuitum group.
The majority of isolates was resistant to 2-4 antibiotics. Multiresistant strains occurred also in a control soil that has a long history
without the exposure to antibiotic-containing manure. Seven isolates grew at 37°C, including the species M. septicum and M. fortuitum
known for infections in humans. This study shows that multiresistant RGM close to known human pathogens occur in grassland soils
regardless the soil history of manuring.

The thought that tuberculosis and the mycobacteria could cause diabetes seems farfetched, but is not. The peculiar relationship and frequent association of diabetes mellitus and tuberculosis has been observed for more than 2000 years, yet the reason for this correlation is, to this day, not known. Before the discovery of insulin, a diagnosis of diabetes was a death sentence within 5 years, and the usual cause of that death was tuberculosis. Despite this, in the 5th century, tuberculosis was already being portrayed as a "complication" of diabetes, a view little changed to this day, parroting Root's original 1934 description of "a one-sided relationship": tuberculosis still seen as a common complication of diabetes, while diabetes is thought to be no more common among TB patients than in the population at large. To Nichol's, this was "not logically tenable" and in his study of 178 otherwise healthy, non-diabetic military men with tuberculosis at Fitzsimmons Army Hospital, one-third had abnormal glucose screening tests. But despite his findings and those of Reaud in New York and others, this was not being recognized elsewhere, and Nichols wanted to know why. Nichols concluded that the incidence of diabetes among tuberculosis patients was considerably underestimated and that in tuberculosis patients, diabetes develops quite commonly. Diabetes was easy to detect. Tuberculosis and the mycobacteria were not. The evidence for a mycobacterial cause of diabetes is mounting rapidly. Schwartz and Haas both linked Type-2 diabetes to tuberculosis. And the pancreatic islet amyloid deposits that they found as a by-product of systemic tubercular infection have recently been dissolved by rifampicin, a first line drug against tuberculosis. Engelbach spoke of "transitory" diabetes in TB and Karachunskii noted changes in carbohydrate metabolism in patients with tuberculosis which commonly led to insulin deficiency with persistent hyperglycemia. Furthermore, mycobacterial elements have been shown recently not only to cause "autoimmune" Type-1 diabetes in NOD (non-obese diabetic) mice, but act as a vaccine to stop the inevitable diabetes that would otherwise materialize. The documentation of patient cases where TB has preceded and come before the development of diabetes is extensive yet underplayed and both Lin's and Tsai's studies speak of tuberculosis complicated by diabetes. Diabetes has been around since the first century AD, in a perpetual state of coping and managing. It is time, it is long past time, to cure diabetes. But current models as to its cause are not equipping us to do so.

Microbial survival in dynamic environments requires the ability to successfully respond to abrupt changes in osmolarity. The mechanosensitive channel of large conductance (MscL) is a ubiquitous channel that facilitates the survival of bacteria and archaea under severe osmotic downshock conditions by relieving excess turgor pressure in response to increased membrane tension. A prominent structural feature of MscL, the cytoplasmic C-terminal domain, has been suggested to influence channel assembly and function. In this report, we describe the X-ray crystal structure and electrophysiological properties of a C-terminal domain truncation of the Mycobacterium tuberculosis MscL (MtMscLΔC). A crystal structure of MtMscLΔC solubilized in the detergent n-dodecyl-β-D-maltopyranoside reveals the pentameric, closed state-like architecture for the membrane spanning region observed in the previously solved full-length MtMscL. Electrophysiological characterization demonstrates that MtMscLΔC retains mechanosensitivity, but with conductance and tension sensitivity more closely resembling full length EcMscL than MtMscL. This study establishes that the C-terminal domain of MtMscL is not required for oligomerization of the full-length channel, but rather influences the tension sensitivity and conductance properties of the channel. The collective picture that emerges from these data is that each MscL channel structure has characteristic features, highlighting the importance of studying multiple homologs. Mechanosensitive (MS) channels transduce mechanical stimuli into a variety of cellular responses. MS channels are widely distributed through all kingdoms of life, including bacteria and archaea, where they play a prominent role in maintaining proper osmoregulation 1-5. Sudden changes in osmolarity, such as osmotic downshock during exposure to hypotonic conditions, increases turgor pressure across the cell membrane. This in turn increases the tension within the lipid bilayer and activates a network of MS channels which respond by opening typically nonselective pores in the membrane to release cell contents and alleviate the pressure 1,5-7. Bacterial MS channels are characterized by their relatively high and non-selective conductances, ranging from ~0.1 nS to ~3 nS under defined experimental conditions 6-8 , which is up to several orders of magnitude greater than typical for ion-selective channels 3,6,9. Of these channels, the mechanosensitive channel of large conductance (MscL) exhibits the largest conductance, with an estimated open state pore diameter of ~25-35 Å-large enough for the passage of molecules up to 9 kDa 4,7,10,11. In view of the high conductance yet small subunit size of MscL (Escherichia coli MscL (EcMscL) has 136 residues; Fig. 1), the oligomeric nature of this channel was appreciated from its initial discovery 11. Considerable efforts have been made to establish the molecular mechanism of MscL gating 3,12 , employing a variety of techniques including electrophysiology, biochemical and genetic studies, molecular dynamics

Se comunican dos casos clínicos poco frecuentes de micobacteriosis cutáneas; uno adquirido en la comunidad y uno en medio hospitalario, ambos pacientes del género masculino, con dermatosis crónicas constituidas por úlceras, fístulas y placas de aspecto papilomatoso. El diagnóstico fue tardío y la evolución no fue satisfactoria, a pesar de prescribir tratamiento adecuado contra Mycobacterium chelonae. El propósito de este artículo es mostrar la manifestación clínica de las lesiones para sospecha diagnóstica temprana y oportuna de las diferentes disciplinas médicas que participan en la atención de los pacientes con estas infecciones.

This paper reports the two rare clinical cases of cutaneous mycobacterial infections. One of them acquired the infection within the community while the other was a noso-comial case, both cases were male with chronic ulcers, fistulae and papilloma-like lesions. In both cases diagnosis was late and evolution, despite correct Mycobacterium chelonae treatment, was unsatisfactory. The purpose of this paper is to show the clinical presentation and accurate suspicion of infection of the different medical areas involved in its management.

Mycobacterium avium subsp. paratuberculosis (MAP) is the bacterium that causes Johne’s disease in cattle. Although infected cattle can be identified by examining fecal, blood, or milk samples, the gold standard is identification of MAP in tissue samples postmortem. Although tissue samples are commonly frozen, the ability to detect MAP in frozen–thawed tissue samples has apparently not been reported. We therefore determined the ability to detect MAP in tissue samples following freezing. Tissue samples were collected from calves that were either inoculated (IN) 3 mo prior, or contact-exposed (CE) for 3 mo. Following autopsy, tissues were immediately processed for culture, followed by DNA extraction and detection by qPCR. Samples were categorized as positive or negative based on the cycle threshold (Ct) value. The remaining unprocessed tissue samples were frozen at −80°C. After 18 mo, 50 tissue samples designated MAP-positive were thawed and processed for detection of MAP. Four (8%) sa...

Tuberculosis is an old infectious bacterial disease caused by Mycobacterium tuberculosis that was once considered lethal having no treatment but now with new inventions treatment is possible. Worldwide, the incidence of Tuberculosis is falling 2% per year but still it is a major cause of mortality killing 2-3 million people every year. Pakistan ranks 5th globally with 5.1% of total national disease burden. A retrospective study was executed at District tuberculosis hospital Mansehra to determine the trends of Tuberculosis. A total of 2619 patients were reported in three years (2015-2017), where 1389 (53%) were females and 1230 (47%) were males. Large number of cases were observed from tehsil Mansehra (76.4%) followed by Baffa, Balakot, and Oghi. The age of 1-19 years (39.4%) was found to be highly susceptible for tuberculosis. Chances for recurrence of Tuberculosis were notably reduced shown by proportion of new patients (91%) greater than relapse. Through years maximum number of tuberculosis cases were treated (63.5%) while minimum proportion was observed for mortality (2%) and treatment failure (1%). Over years, the incidence of pulmonary tuberculosis (75.7%) was increased as compared to extra pulmonary tuberculosis (24.3%). Highest numbers of tuberculosis cases were reported in summer (28.7%).

Keywords:  Tuberculosis (TB), Mycobacterium Tuberculosis, trends, gender, Tehsil, age, Pulmonary TB, Extra Pulmonary TB, seasonality

Mycobacterium ulcerans, the causative agent of Buruli ulcer, is the third most common mycobacterial disease after tuberculosis and leprosy. The present treatment options are limited and emergence of treatment resistant isolates represents a serious concern and a need for better therapeutics. Conventional drug discovery methods are time consuming and labor-intensive. Unfortunately, the slow growing nature of M. ulcerans in experimental conditions is also a barrier for drug discovery and development. In contrast, recent advancements in complete genome sequencing, in combination with cheminformatics and computational biology, represent an attractive alternative approach for the identification of therapeutic candidates worthy of experimental research. A computational, comparative genomics workflow was defined for the identification of novel therapeutic candidates against M. ulcerans, with the aim that a selected target should be essential to the pathogen, and have no homology in the human host. Initially, a total of 424 genes were predicted as essential from the M. ulcerans genome, via homology searching of essential genome content from 20 different bacteria. Metabolic pathway analysis showed that the most essential genes are associated with carbohydrate and amino acid metabolism. Among these, 236 proteins were identified as non-host and essential, and could serve as potential drug and vaccine candidates. Several drug target prioritization parameters including druggability were also calculated. Enzymes from several pathways are discussed as potential drug targets, including those from cell wall synthesis, thiamine biosynthesis, protein biosynthesis, and histidine biosynthesis. It is expected that our data will facilitate selection of M. ulcerans proteins for successful entry into drug design pipelines.

The present study was designed to determine the half-life of gfp m 2+ mRNA, which encodes mycobacterial codon-optimised highly fluorescent GFP m 2+ protein, and to find out whether mycobacterial promoter activity can be quantitated more accurately using the mRNA levels of the reporter gene, gfp m 2+ , than the fluorescence intensity of the GFP m 2+ protein. Quantitative PCR of gfp m 2+ mRNA in the pulse-chased samples of the rifampicin-treated Mycobacterium smegmatis/gfp m 2+ transformant showed the half-life of gfp m 2+ mRNA to be 4.081 min. The levels of the gfp m 2+ mRNA and the fluorescence intensity of the GFP m 2+ protein, which were expressed by the promoters of Mycobacterium tuberculosis cell division gene, ftsZ (MtftsZ), were determined using quantitative PCR and fluorescence spectrophotometry, respectively. The data revealed that quantification of mycobacterial promoter activity by determining the gfp m 2+ mRNA levels is more accurate and statistically significant than the measurement of GFP m 2+ fluorescence intensity, especially for weak promoters.

Mycobacterium avium subsp. paratuberculosis (MAP) is the bacterium that causes Johne's disease in cattle. Although infected cattle can be identified by examining fecal, blood, or milk samples, the gold standard is identification of MAP in tissue samples postmortem. Although tissue samples are commonly frozen, the ability to detect MAP in frozenthawed tissue samples has apparently not been reported. We therefore determined the ability to detect MAP in tissue samples following freezing. Tissue samples were collected from calves that were either inoculated (IN) 3 mo prior, or contact-exposed (CE) for 3 mo. Following autopsy, tissues were immediately processed for culture, followed by DNA extraction and detection by qPCR. Samples were categorized as positive or negative based on the cycle threshold (Ct) value. The remaining unprocessed tissue samples were frozen at −80°C. After 18 mo, 50 tissue samples designated MAP-positive were thawed and processed for detection of MAP. Four (8%) samples were qPCR-negative, and Ct values of the remaining 46 samples were higher after freezing. Given the small numerical change in Ct values for MAP-positive samples after 18 mo of frozen storage, freezing and thawing may have had some deleterious effects on MAP detection in tissues. Although the decrease in ability to detect MAP-positive samples was minor for IN calves, there may be a greater effect for CE calves that should be considered when freezing tissue samples.

Insulin resistance (IR) is a key determinant of type 2 diabetes (T2D) and other metabolic disorders. This genome-wide association study (GWAS) was designed to shed light on the genetic basis of fasting insulin (FI) and IR in 927 non-diabetic African Americans. 5 396 838 single-nucleotide polymorphisms (SNPs) were tested for associations with FI or IR with adjustments for age, sex, body mass index, hypertension status and first two principal components. Genotyped SNPs (n 5 12) with P < 5 3 10 26 in African Americans were carried forward for de novo genotyping in 570 non-diabetic West Africans. We replicated SNPs in or near SC4MOL and TCERG1L in West Africans. The meta-analysis of 1497 African Americans and West Africans yielded genome-wide significant associations for SNPs in the SC4MOL gene: rs17046216 (P 5 1.7 3 10 28 and 2.9 3 10 28 for FI and IR, respectively); and near the TCERG1L gene with rs7077836 as the top scoring (P 5 7.5 3 10 29 and 4.9 3 10 210 for FI and IR, respectively). In silico replication in the MAGIC study (n 5 37 037) showed weak but significant association (adjusted P-value of 0.0097) for rs34602777 in the MYO5A gene. In addition, we replicated previous GWAS findings for IR and FI in Europeans for GCKR, and for variants in four T2D loci (FTO, IRS1, KLF14 and PPARG) which exert their action via IR. In summary, variants in/near SC4MOL, and TCERG1L were associated with FI and IR in this cohort of African Americans and were replicated in West Africans. SC4MOL is under-expressed in an animal model of T2D and plays a key role in lipid biosynthesis, with implications for the regulation of energy metabolism, obesity and dyslipidemia. TCERG1L is associated with plasma adiponectin, a key modulator of obesity, inflammation, IR and diabetes.

Se denominan micobacterias atípicas todas las especies del gé nero Mycobacterium capaces de producir enfermedades en el hombre, excepto las que forman el complejo M. tuberculosis y M. leprae. Aunque el té rmino «atípicas» ha sido muy criticado y se han propuesto diversas alternativas, como micobacterias ambientales, micobacterias anó nimas y micobacterias no tuberculosas (MNT), el hecho cierto es que sigue siendo ampliamente utilizado en la literatura mé dica y en la prá ctica clínica 1 .

Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be one of the primary sources of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-sectional analysis of longitudinally collected samples on 3 dairy farms. Composite samples from multiple environmental sites in 3 commercial dairy herds in the Northeast US were cultured quarterly for MAP, providing 1131 samples (133 (11.8%) were culture-positive), and all adult animals in the herds were tested biannually by fecal culture (FC), for 6 years. Of the environmental sites sampled, manure storage areas and shared alleyways were most likely to be culture-positive. Environmental sample results were compared to FC results from either the concurrent or previous sampling date at both the herd and the pen level. At the herd level, a 1 log unit increase in average fecal shedding increased the odds of a positive non-pen environmental sample by a factor of 6 and increased the average amount of MAP in non-pen samples by 2.9 cfu/g. At the pen level, a 1 log unit increase in average fecal shedding in the pen increased the odds of a positive environment by a factor of 2.4 and the average amount of MAP was increased by 3.5 cfu/g. We were not able to model the relationship between non-pen environmental sample status and the distance between shedding animals and the sample's location, and neighboring pens did not significantly affect the results of the pen-level analysis. The amount of MAP in pen-level samples and the probability of a pen testing positive for MAP were both positively but non-significantly correlated with the number of animals in the pen shedding >30 cfu/g of MAP. At least 6 environmental samples met the criteria for the U.S. Voluntary Bovine Johne's Disease Control Program on 47 of the 72 sampling dates; of these, 19 of the 47 FC-positive sampling dates were positive by the 6-sample environmental testing method, resulting in a herd sensitivity of 0.40 (95% CI: 0.26-0.54). None of the 3 FC-negative sampling dates produced positive environmental samples. Although environmental sampling can be * Corresponding author at: S2-064 Schurman Hall,

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for the identification of nontuberculous mycobacterial (NTM) isolates was evaluated in this study. Overall, 125 NTM isolates were analyzed by MALDI-TOF and GenoType CM/AS. Identification by 16S rRNA/hsp65 sequencing was considered the gold standard. Agreements between MALDI-TOF and GenoType CM/AS with the reference method were, respectively, 94.4% and 84.0%. In 17 cases (13.6%), results provided by GenoType and MALDI-TOF were discordant; however, the reference method agreed with MALDI-TOF in 16/17 cases (94.1%; P = 0.002).

The authors describe a study of the tuberculosis (TB) incidence in cattle exposed to low doses of radiation resulting from the Chernobyl (pronounced ‘Chornobyl’ in Ukrainian) nuclear plant catastrophe in 1986. The purpose of the study was to determine if ionising radiation influences the number of outbreaks of bovine TB and their severity on farms in the Kyiv, Cherkasy and Chernigiv regions of Ukraine. These farms are all located within a 200 km radius of Chernobyl and have had low-dose radiation pollution. Pathological and blood samples were taken from cattle in those regions that had positive TB skin tests. Mycobacterium spp. were isolated, differentiated by PCR, analysed and tested in guinea-pigs and rabbits. Species differentiation showed a significant percentage of atypical mycobacteria, which resulted in the allergic reactions to tuberculin antigen in the skin test. Mixed infection of M. bovis and M. avium subsp. hominissuis was found in three cases. The results concluded that low-dose radiation plays a major role in the occurrence of bovine TB in regions affected by the Chernobyl nuclear disaster.

Collagen deposition within granulomas formed after Mycobacterium avium infection was analysed on histological sections stained with Masson’s trichrome using acquired computerized image analysis and a program that was specifically designed for that purpose. Comparison was made between immunocompetent C57BL⁄ 6 mice and mice genetically deficient in the inducible nitric oxide (NO) synthase gene (iNOS– ⁄ – mice) infected with either a highly virulent strain or a moderately virulent strain of M. avium. iNOS-deficient mice were more resistant to the highly virulent strain than control C57B1⁄ 6 mice, but both strains were equally susceptible to the less virulent M. avium strain. Collagen distribution in the granuloma was found in the cuff surrounding the granuloma in an area rich in lymphoid cells as well as inside the granuloma itself, conferring a mesh-like structure within that lesion. It was seen that iNOS– ⁄ – mice induced a higher collagen deposition than C57BL ⁄ 6 mice and that such collagen deposition varied with the mycobacterial strain used to
infect the animals.

The growth in C57Bl/6 mice of seven distinct Mycobacterium avium strains was not exacerbated by the disruption of the inducible (type 2) nitric oxide synthase regardless of the virulence of the strain. The susceptibility of this panel of M. avium strains to reactive nitrogen intermediates in a cell-free system, namely the exposure to acidified nitrite or to the NO donor NOC-18, showed that there is no correlation between strain virulence and the corresponding minimal bactericidal and minimal inhibitory concentrations for those compounds determined in vitro.

Environmental contamination with Mycobacterium avium subsp. paratuberculosis (MAP) is thought to be one of the primary sources of infection for dairy cattle. The exact link between fecal shedding of MAP by individual cows and environmental contamination levels at the herd level was explored with a cross-sectional analysis of longitudinally collected samples on 3 dairy farms. Composite samples from multiple environmental sites in 3 commercial dairy herds in the Northeast US were cultured quarterly for MAP, providing 1131 samples (133 (11.8%) were culture-positive), and all adult animals in the herds were tested biannually by fecal culture (FC), for 6 years. Of the environmental sites sampled, manure storage areas and shared alleyways were most likely to be culture-positive. Environmental sample results were compared to FC results from either the concurrent or previous sampling date at both the herd and the pen level. At the herd level, a 1 log unit increase in average fecal shedding increased the odds of a positive non-pen environmental sample by a factor of 6 and increased the average amount of MAP in non-pen samples by 2.9 cfu/g. At the pen level, a 1 log unit increase in average fecal shedding in the pen increased the odds of a positive environment by a factor of 2.4 and the average amount of MAP was increased by 3.5 cfu/g. We were not able to model the relationship between non-pen environmental sample status and the distance between shedding animals and the sample's location, and neighboring pens did not significantly affect the results of the pen-level analysis. The amount of MAP in pen-level samples and the probability of a pen testing positive for MAP were both positively but non-significantly correlated with the number of animals in the pen shedding >30 cfu/g of MAP. At least 6 environmental samples met the criteria for the U.S. Voluntary Bovine Johne's Disease Control Program on 47 of the 72 sampling dates; of these, 19 of the 47 FC-positive sampling dates were positive by the 6-sample environmental testing method, resulting in a herd sensitivity of 0.40 (95% CI: 0.26-0.54). None of the 3 FC-negative sampling dates produced positive environmental samples. Although environmental sampling can be * Corresponding author at: S2-064 Schurman Hall,

Analogues of the antibiotic thiolactomycin (TLM) have been synthesized and have been shown to have enhanced activity against whole cells of Mycobacterium tuberculosis H37Rv and against mycolic acid biosynthesis in cell extracts of Mycobacterium smegmatis. TLM has a methyl-branched butadienyl side chain attached at position 5 on a &#39;thiolactone&#39; ring, namely 4-hydroxy-3,5-dimethyl-5H-thiophen-2-one. Various combinations of strong bases were explored to create a reactive anion at position 5 on the thiolactone ring which could react with halides to produce 5-substituted derivatives; the best reagent was two equivalents of lithium-bis-(trimethylsilyl)amide in tetrahydrofuran. The analogue with a 5-tetrahydrogeranyl substituent showed the best biological activity with an MIC(90) for M. tuberculosis of 29 micro M and 92% mycolate inhibition in extracts of M. smegmatis, as compared to 125 micro M and 54%, respectively, for TLM; other related C(10) and C(15) isoprenoid derivatives ha...

Johne&#39;s disease, a chronic enteritis caused by Mycobacterium avium ssp. paratuberculosis (MAP), causes large economic losses to the dairy industry worldwide. Fecal shedding of MAP contaminates the environment, feed, and water and contributes to new infections on farm, yet there is limited knowledge regarding mechanisms of shedding, extent of intermittent shedding, and numbers of MAP bacteria shed. The objectives were to (1) compare (in an experimental setting) the frequency at which intermittent shedding occurred and the quantity of MAP shed among pen mates that were inoculated or contact-exposed (CE); and (2) determine whether an association existed between inoculation dose and quantity of MAP shed. In the first experiment, 32 newborn Holstein-Friesian bull calves were allocated to pens in groups of 4, whereby 2 calves were inoculated with a moderate dose (MD; 5 × 108 cfu) of MAP and 2 calves acted as CE. Calves were group-housed for 3 mo, fecal samples were collected and cultu...

Background: Buruli ulcer is the third most common mycobacterial infection worldwide. It is endemic in tropical, subtropical, and temperate climates. It causes devastating disease with morbidity and mortality. The treatment duration is long and the regimens considered are limited. Chronic cutaneous ulcers of mycobacterial etiology have been reported previously in Amman, but these were not associated with Mycobacterium ulcerans infection. Methods: The case patient's initial diagnosis was based on chronological and morphological features, combined with appropriate diagnostic tests. The skin features were assessed histopathologically. Skin testing was positive for acid-fast bacilli (AFB), and M. ulcerans was identified by DNA strip test (GenoType Mycobacterium CM/AS, Hain Lifescience), which is based on a PCR technique targeting a 23S rRNA gene region, followed by reverse hybridization and a line probe technology. Results: The skin mycobacterial infection was evaluated and verified as having a Mycobacterium marinum-M. ulcerans pattern in the GenoType CM assay. It was then counted as a pattern representing individual species and was resolved with the GenoType AS assay as having an M. ulcerans pattern. M. ulcerans DNA was isolated and amplified by PCR, and then detected against reverse hybridization probes in the strip assay. Conclusions: An indigenous case of M. ulcerans (Buruli ulcer) is reported for the first time from Jordan and the surrounding region.

Autophagy represents a key pathway in innate immune defense to restrict Mycobacterium tuberculosis growth inside host macrophages. Induction of autophagy has been shown to promote mycobacterial phagosome acidification and acquisition of lysosomal hydrolases, resulting in the elimination of intracellular M. tuberculosis reference strains such as H37Rv. The notorious Beijing genotype has been previously shown to be hyper-virulent and associated with increased survival in host cells and a high mortality rate in animal models, but the underlying mechanism that renders this family to have such advantages remains unclear. We hypothesize that autophagic control against M. tuberculosis Beijing strains may be altered. Here, we discovered that the Beijing strains can resist autophagic killing by host cells compared with that of the reference strain H37Rv and a strain belonging to the East African Indian genotype. Moreover, we have determined a possible underlying mechanism and found that the ...

Mycobacterium kansasii disease present epidemiological, clinical and radiological features similar to Mycobacterium tuberculosis. Mycobacterium kansasii is the second most frequent mycobacteria isolated in human immunodeficiency virus infected patients. Confirmed diagnosis is often difficult according with the American Thoracic Society criteria. Here we describe a patient with AIDS that developed a severe lung compromise due to Mycobacterium kansasii. Epidemiological, clinical, radiological and diagnosis methods are analyzed.

Restless legs syndrome (RLS) is a sensorimotor disorder with an age-dependent prevalence of up to 10% in the general population above 65 years of age. Affected individuals suffer from uncomfortable sensations and an urge to move in the lower limbs that occurs mainly in resting situations during the evening or at night. Moving the legs or walking leads to an improvement of symptoms. Concomitantly, patients report sleep disturbances with consequences such as reduced daytime functioning. We conducted a genome-wide association study (GWA) for RLS in 922 cases and 1,526 controls (using 301,406 SNPs) followed by a replication of 76 candidate SNPs in 3,935 cases and 5,754 controls, all of European ancestry. Herein, we identified six RLS susceptibility loci of genome-wide significance, two of them novel: an intergenic region on chromosome 2p14 (rs6747972, P = 9.03 6 10 211 , OR = 1.23) and a locus on 16q12.1 (rs3104767, P = 9.4 6 10 219 , OR = 1.35) in a linkage disequilibrium block of 140 kb containing the 59-end of TOX3 and the adjacent non-coding RNA BC034767.

Interleukin-10 is an immunomodulatory cytokine that has been implicated, along with IFN-c, in the disease sequelae of mycobacterial infection. In order to investigate the role of IL-10 in marsupial disease models we sequenced and characterised the IL10 gene in the tammar wallaby (Macropus eugenii) and rufous hare-wallaby (Lagorchestes hirsutus). An isoform IL-10D3, in which an in-frame deletion of exon 3 occurs, was discovered in both macropod species. Analysis of wallaby and other reported marsupial IL-10 homologs suggests that while marsupial IL-10 is comparable to that of human IL-10, the predicted IL-10D3 protein may play a more complicated role in the modulation of IL-10-directed responses. Expression of the canonical gene and splicing variant was confirmed in both wallabies, and the rufous hare-wallaby showed differential expression across lymph node, spleen and liver, with isoform expression detected in the lymph node. This characterisation and expression of IL-10 in de novo tissues provides a basis for further study into the role of IL-10 in disease models in marsupials.

Insulin resistance (IR) is a key determinant of type 2 diabetes (T2D) and other metabolic disorders. This genome-wide association study (GWAS) was designed to shed light on the genetic basis of fasting insulin (FI) and IR in 927 non-diabetic African Americans. 5 396 838 single-nucleotide polymorphisms (SNPs) were tested for associations with FI or IR with adjustments for age, sex, body mass index, hypertension status and first two principal components. Genotyped SNPs (n 5 12) with P < 5 3 10 26 in African Americans were carried forward for de novo genotyping in 570 non-diabetic West Africans. We replicated SNPs in or near SC4MOL and TCERG1L in West Africans. The meta-analysis of 1497 African Americans and West Africans yielded genome-wide significant associations for SNPs in the SC4MOL gene: rs17046216 (P 5 1.7 3 10 28 and 2.9 3 10 28 for FI and IR, respectively); and near the TCERG1L gene with rs7077836 as the top scoring (P 5 7.5 3 10 29 and 4.9 3 10 210 for FI and IR, respectively). In silico replication in the MAGIC study (n 5 37 037) showed weak but significant association (adjusted P-value of 0.0097) for rs34602777 in the MYO5A gene. In addition, we replicated previous GWAS findings for IR and FI in Europeans for GCKR, and for variants in four T2D loci (FTO, IRS1, KLF14 and PPARG) which exert their action via IR. In summary, variants in/near SC4MOL, and TCERG1L were associated with FI and IR in this cohort of African Americans and were replicated in West Africans. SC4MOL is under-expressed in an animal model of T2D and plays a key role in lipid biosynthesis, with implications for the regulation of energy metabolism, obesity and dyslipidemia. TCERG1L is associated with plasma adiponectin, a key modulator of obesity, inflammation, IR and diabetes.

Leukocyte migration into the epithelial compartment is an important feature in the active phase of mycobacterial infections. In this study, we used the Transwell model to investigate the mechanisms behind mycobacteria-induced leukocyte recruitment and investigated the role of TLR2 and TLR4 in this process. Infection of epithelial cells resulted in significantly increased secretion of the neutrophil chemotactic CXCL8 and IL-6, but no secretion of monocyte chemotactic CCL2 or TNF-a was observed. In contrast to epithelial response, mycobacteria-infected neutrophils and monocytes secreted all these cytokines. Corresponding with epithelial cytokine response, mycobacterial infection of the epithelial cells increased neutrophil diapedesis, but decreased monocyte recruitment. However, monocyte recruitment towards mycobacteria infected epithelial cells significantly increased following addition of neutrophil pre-conditioned medium. Mycobacterial infection also increases alveolar epithelial expression of TLR2, but not TLR4, as analyzed by flow cytometry, Western blotting and visualized by confocal microscopy. Blocking of TLR2 inhibited neutrophil recruitment and cytokine secretion, while blocking of TLR4 had a lesser effect. To summarize, we found that primary alveolar epithelial cells produced a selective TLR2-dependent cytokine secretion upon mycobacterial infection. Furthermore, we found that cooperation between cells of the innate immunity is required in mounting proper antimicrobial defence.