Continuous and Discontinuous Cigarette Smoke Exposure Differentially Affects Protective Th1 Immunity against Pulmonary Tuberculosis

Pulmonary tuberculosis (TB), caused by Mycobacterium tuberculosis, is the leading cause of death due to a bacterial pathogen. Emerging epidemiologic evidence suggests that the leading risk factor associated with TB mortality is cigarette smoke exposure. Despite this, it remains poorly understood what is the effect of cigarette smoke exposure on anti-TB immunity and whether its potential detrimental effect can be reversed by cigarette smoking cessation. In our current study, we have investigated the impact of both continuous and discontinuous cigarette smoke exposure on the development of anti-mycobacterial type 1 immunity in murine models. We find that while continuous cigarette smoke exposure severely impairs type 1 immunity in the lung, a short-term smoking cessation allows rapid restoration of anti-mycobacterial immunity. The ability of continuous cigarette smoke exposure to dampen type 1 protective immunity is attributed locally to its affects on innate immune cells in the lung. Continuous cigarette smoke exposure locally, by not systemically, impairs APC accumulation and their production of TNF, IL-12, and RANTES, blunts the recruitment of CD4+IFN-γ+ T cells to the lung, and weakens the formation of granuloma. On the other hand, smoking cessation was found to help restore type 1 immunity by rapidly improving the functionality of lung APCs, enhancing the recruitment of CD4+IFN-γ+ T cells to the lung, and promoting the formation of granuloma. Our study for the first time demonstrates that continuous, but not discontinuous, cigarette smoke exposure severely impedes the lung expression of anti-TB Th1 immunity via inhibiting innate immune activation and lung T cell recruitment. Our findings thus suggest cigarette smoking cessation to be beneficial to the control of pulmonary TB.     

Production of the Streptomyces scabies coronafacoyl phytotoxins involves a novel biosynthetic pathway with an F420‐dependent oxidoreductase and a short‐chain dehydrogenase/reductase

Coronafacoyl phytotoxins are secondary metabolites that are produced by various phytopathogenic bacteria, including several pathovars of the Gram‐negative bacterium Pseudomonas syringae as well as the Gram‐positive potato scab pathogen Streptomyces scabies. The phytotoxins are composed of the polyketide coronafacic acid (CFA) linked via an amide bond to amino acids or amino acid derivatives, and their biosynthesis involves the cfa and cfa‐like gene clusters that are found in P. syringae and S. scabies, respectively. The S. scabies cfa‐like gene cluster was previously reported to contain several genes that are absent from the P. syringae cfa gene cluster, including one (oxr) encoding a putative F₄₂₀—dependent oxidoreductase, and another (sdr) encoding a predicted short‐chain dehydrogenase/reductase. Using gene deletion analysis, we demonstrated that both oxr and sdr are required for normal production of the S. scabies coronafacoyl phytotoxins, and structural analysis of metabolites that accumulated in the Δsdr mutant cultures revealed that Sdr is directly involved in the biosynthesis of the CFA moiety. Our results suggest that S. scabies and P. syringae use distinct biosynthetic pathways for producing coronafacoyl phytotoxins, which are important mediators of host‐pathogen interactions in various plant pathosystems.     

Isolation and PCR detection of <Emphasis Type="Italic">Enterobacter sakazakii</Emphasis> in South African food products,specifically infant formula milks

Enterobacter sakazakii has recently been identified as an opportunistic pathogen. The current culture-dependent detection methods for these bacteria are time-consuming and in this study a PCR method for the detection of E. sakazakii in South African food products, including an internal amplification control (IAC) was developed. DNA was isolated and amplified from the products and they were plated on selective growth media after pre-enrichment and enrichment in Enterobacteriaceae enrichment broth. Four of the 22 products tested positive for the presence of E. sakazakii, confirmed by PCR detection and growth on selective media. The PCR method proved effective in detecting E. sakazakii in South African products after three days and could serve as an alternative for traditional microbiological techniques.     

Cell-mediated Immune Responses in Chronic Liver Diseases

Studies of the cell-mediated response to liver antigens, using the leucocyte migration test, in 163 patients with various liver disorders showed that abnormal responses were almost confined to active chronic hepatitis (53% abnormal), primary biliary cirrhosis (64%), and cryptogenic cirrhosis (29%). The test was also abnormal in five out of seven patients with jaundice due to drug hypersensitivity and in one patient with acute infectious hepatitis at a time when mitochondrial antibodies were present in the serum. More of those with active chronic hepatitis on prednisone or azathioprine had normal tests than of those who were untreated, and in 8 out of 10 examined serially during therapy there was an accompanying improvement in leucocyte migration. Abnormal responses to salivary gland or kidney antigens were also found in nearly half of those with features of Sjögren''s syndrome or renal tubular acidosis as part of a multisystem involvement—this, though occurring in cryptogenic cirrhosis, was found with greater frequency in active chronic hepatitis and primary biliary cirrhosis. These cell-mediated immune responses, perhaps triggered by the initial damage to the liver from viral or other agents, may be responsible both for the perpetuation of the liver disease and, because of common surface antigens, for the damage to other organs.     

Development and assessment of a landscape-scale ecological threat index for the Lower Colorado River Basin

Anthropogenic disturbances impact freshwater biota but are rarely incorporated into conservation planning due to the difficulties in quantifying threats. There is currently no widely accepted method to quantify disturbances, and determining how to measure threats to upstream catchments using disturbance metrics can be time consuming and subjective. We compared four watershed-scale ecological threat indices for the Lower Colorado River Basin (LCRB) using landscape-level threats of land use (e.g., agricultural and urban lands), waterway development and diversions (e.g., number of canals, dams), and human development (e.g., road and railroads density, pollution sites). The LCRB is an ideal region to assess ecological threat indices because of the increasing need for conservation to ensure the persistence of native fishes in highly altered habitat. Each threat was measured for severity (i.e., level of influence on the upstream watershed) and frequency throughout each watershed; both severity and frequency were measured using two different methods. Severity values were based either on peer-reviewed literature and weighted in accordance to their published ecological impact, or assumed equal severity across stressors. Threat frequency was calculated according to either the presence/absence of each stressor, or on the relative density of each stressor in the watershed. Each measure of severity was combined with a measure of frequency, creating four ecological threat indices, and transformed to a 0–100 scale. Threat indices were highly correlated (slopes of 0.94–1.63; R² of 0.82–0.98), and were highest for watersheds close to urban centers, including Phoenix, Tucson, and Flagstaff, Arizona, and Las Vegas, Nevada. Road crossings and density appeared to be the most influential stressors in the index, but the removal of any individual stressor only changed the index by <5.1 units. Our results indicate that a simpler index with less subjectivity (i.e., presence/absence of a stressor in a watershed) provides similar results to the more subjective measure of threats (i.e., peer-reviewed threat severity). Because these threats have been linked to ecological health, the development of the index should be a useful tool to identify regions of greatest potential threat to aquatic biota and can aid in conservation planning for the Lower Colorado River Basin.