The effectiveness of BCG vaccination in preventing Mycobacterium tuberculosis infection and disease in Greenland.

BACKGROUND: The BCG vaccine's ability to prevent Mycobacterium tuberculosis infection (MTI) remains highly debated. In Greenland, BCG vaccination was introduced in 1955, but was temporarily discontinued (1991-1996) due to nationwide policy changes. The study aimed to use the transient stop in BCG vaccination to evaluate the effect of vaccination on MTI prevalence and TB incidence. METHODS: MTI study: A cross-sectional study (2012), comprising East Greenlanders born during 1982-2006, evaluated the effect of BCG vaccination on MTI prevalence; a positive interferon γ release assay defined an MTI case. Associations were estimated using logistic regression. TB study: a cohort study covering the same birth cohorts with follow-up until 2012 evaluated the vaccine's effect on TB incidence. A personal identifier allowed for follow-up in the TB notification system. Associations were estimated using Cox regression. RESULTS: MTI study: Included 953 participants; 81% were BCG-vaccinated; 29% had MTI, 23% among vaccinated and 57% among non-vaccinated. BCG vaccination reduced the odds of MTI, OR 0.52 (95% CI 0.32 to 0.85), p=0.01. Vaccine effectiveness against MTI was 20%. TB study: Included 1697 participants followed for 21,148 person-years. 6% were notified with TB, 4% among vaccinated and 11% among non-vaccinated. BCG vaccination reduced the risk of TB, HR 0.50 (95% CI 0.26 to 0.95), p=0.03, yielding a vaccine effectiveness of 50%. CONCLUSIONS: BCG vaccination was effective in reducing both MTI and TB disease among children and young adults in a TB high-endemic setting in Greenland.

Bonds and bytes: The odyssey of structural biology.

Characterizing structural and dynamic properties of proteins and large macromolecular assemblies is crucial to understand the molecular mechanisms underlying biological functions. In the field of structural biology, no single method comprehensively reveals the behavior of biological systems across various spatiotemporal scales. Instead, we have a versatile toolkit of techniques, each contributing a piece to the overall puzzle. Integrative structural biology combines different techniques to create accurate and precise multi-scale models that expand our understanding of complex biological systems. This review outlines recent advancements in computational and experimental methods in structural biology, with special focus on recent Artificial Intelligence techniques, emphasizes integrative approaches that combine different types of data for precise spatiotemporal modeling, and provides an outlook into future directions of this field.

Using the systematic review methodology to evaluate factors that influence the persistence of influenza virus in environmental matrices.

Understanding factors that influence persistence of influenza virus in an environment without host animals is critical to appropriate decision-making for issues such as quarantine downtimes, setback distances, and eradication programs in livestock production systems. This systematic review identifies literature describing persistence of influenza virus in environmental samples, i.e., air, water, soil, feces, and fomites. An electronic search of PubMed, CAB, AGRICOLA, Biosis, and Compendex was performed, and citation relevance was determined according to the aim of the review. Quality assessment of relevant studies was performed using criteria from experts in virology, disease ecology, and environmental science. A total of 9,760 abstracts were evaluated, and 40 appeared to report the persistence of influenza virus in environmental samples. Evaluation of full texts revealed that 19 of the 40 studies were suitable for review, as they described virus concentration measured at multiple sampling times, with viruses detectable at least twice. Seven studies reported persistence in air (six published before 1970), seven in water (five published after 1990), two in feces, and three on surfaces. All three fomite and five air studies addressed human influenza virus, and all water and feces studies pertained to avian influenza virus. Outcome measurements were transformed to half-lives, and resultant multivariate mixed linear regression models identified influenza virus surviving longer in water than in air. Temperature was a significant predictor of persistence over all matrices. Salinity and pH were significant predictors of persistence in water conditions. An assessment of the methodological quality review of the included studies revealed significant gaps in reporting critical aspects of study design.

Kinetics of UV(254) inactivation of selected viral pathogens in a static system.

AIMS: The objective of this study was to estimate UV(254) inactivation constants for four viral pathogens: influenza virus type A, porcine respiratory and reproductive syndrome virus (PRRSV), bovine viral diarrhoea virus (BVDV) and reovirus. METHODS AND RESULTS: Viruses in culture medium were exposed to one of nine doses of UV(254) and then titrated for infectious virus. Analysis showed that viral inactivation by UV(254) was more accurately described by a two-stage inactivation model vs a standard one-stage inactivation model. CONCLUSIONS: The results provided evidence for the existence of two heterogeneous viral subpopulations among the viruses tested, one highly susceptible to UV(254) inactivation and the other more resistant. Importantly, inactivation constants based on the one-stage inactivation model would have underestimated the UV(254) dose required for the inactivation of these viruses under the conditions of the experiment. SIGNIFICANCE AND IMPACT OF THE STUDY: To improve the accuracy of estimates, it is recommended that research involving the inactivation of micro-organisms evaluates inactivation kinetics using both one-stage and two-stage models. These results will be of interest to persons responsible for microbial agents under laboratory or field conditions.

Mammographic features and screening outcome in a randomized controlled trial comparing digital breast tomosynthesis and digital mammography.

PURPOSE: To compare the distribution of mammographic features among women recalled for further assessment after screening with digital breast tomosynthesis (DBT) versus digital mammography (DM), and to assess associations between features and final outcome of the screening, including immunohistochemical subtypes of the tumour. METHODS: This randomized controlled trial was performed in Bergen, Norway, and included 28,749 women, of which 1015 were recalled due to mammographic findings. Mammographic features were classified according to a modified BI-RADS-scale. The distribution were compared using 95 % confidence intervals (CI). RESULTS: Asymmetry was the most common feature of all recalls, 24.3 % (108/444) for DBT and 38.9 % (222/571) for DM. Spiculated mass was most common for breast cancer after screening with DBT (36.8 %, 35/95, 95 %CI: 27.2-47.4) while calcifications (23.0 %, 20/87, 95 %CI: 14.6-33.2) was the most frequent after DM. Among women screened with DBT, 0.13 % (95 %CI: 0.08-0.21) had benign outcome after recall due to indistinct mass while the percentage was 0.28 % (95 %CI: 0.20-0.38) for DM. The distributions were 0.70 % (95 %CI: 0.57-0.85) versus 1.46 % (95 %CI: 1.27-1.67) for asymmetry and 0.24 % (95 %CI: 0.16-0.33) versus 0.54 % (95 %CI: 0.43-0.68) for obscured mass, among women screened with DBT versus DM, respectively. Spiculated mass was the most common feature among women diagnosed with non-luminal A-like cancer after DBT and after DM. CONCLUSIONS: Spiculated mass was the dominant feature for breast cancer among women screened with DBT while calcifications was the most frequent feature for DM. Further studies exploring the clinical relevance of mammographic features visible particularly on DBT are warranted.

Interaction between C-Tb and PPD given concomitantly in a split-body randomised controlled trial.

SETTING: Seven tuberculosis (TB) clinics in South Africa. OBJECTIVE: As both purified protein derivative (PPD) and a Mycobacterium tuberculosis-specific skin test (C-Tb) contain region of difference 1 (RD1) antigens, we conducted a study to evaluate whether there was any interaction between the two during concomitant and separate administration in patients with newly diagnosed culture-positive TB. DESIGN: Adult patients with active TB (n = 456, 20% human immunodeficiency virus infected) were randomised to receive only C-Tb, only PPD, or concomitant injection of both C-Tb and PPD using the Mantoux technique. Indurations were read after 48-72 h. QuantiFERON®-TB Gold In-Tube (QFT) was performed in tandem. RESULTS: Of the 456 study participants, 154 simultaneously received both C-Tb and PPD, 153 only C-Tb and 149 only PPD. There was no effect of concomitant injection of PPD on the mean C-Tb induration (19 mm, 95%CI 17-22 vs. 18 mm, 95%CI 16-21; P = 0.91). In patients with active TB, C-Tb sensitivity (78%) was similar to PPD (81%) and QFT (84%; excluding 82/429 [19%] indeterminate results). All tests showed reduced sensitivity in participants with CD4 <100 cells/µl. CONCLUSION: In patients with active TB, there was no interaction between C-Tb and PPD during concomitant injection of both agents. Sensitivities were similar for PPD and C-Tb.

Rapid aerosol transmission of Salmonella among turkeys in a simulated holding-shed environment.

Once turkeys arrive at Midwest processing plants, they are usually held in large open-sided sheds for 1 to 4 h, waiting to be unloaded. In hot, humid weather, large fans are used to cool the birds. The resultant air currents distribute a significant amount of dust to the turkeys. The dust created in this environment could be a factor in the number of Salmonella-contaminated turkeys entering slaughter plants. The objective of this study was to determine if rapid transmission of Salmonella in turkeys could occur from exposure to Salmonella-contaminated dust similar to what may be experienced in holding sheds or in other high-dust environments prior to slaughter. In the first experiment, trials of 3 different concentrations of Salmonella (1.2 x 10(9), 2.6 x 10(7), and 2.6 x 10(5) cfu/g) were conducted to determine if transmission of Salmonella enterica serovar Typhimurium var. typhimurium(chi)4232 to turkeys 2 to 4 h after aerosol exposure to contaminated feces is possible. Results showed that turkeys became infected after 2 h of exposure to airborne-contaminated feces with a concentration level of 2.6 x 10(5) cfu of Salmonella Typhimurium/g. In the second experiment, consisting of 3 trials, 1 bank (5 cages wide and 3 cages high) of turkeys (n = 15 birds per trial) was exposed to another bank of cages of S. Typhimurium-inoculated (n = 15) birds for 2 to 4 h using a fan similar to the type in processing-plant cooling sheds. Results from this experiment demonstrated that birds could be contaminated with S. Typhimurium after 2 h of exposure. Results of both studies implicate contaminated dust as a route of rapid airborne transmission of Salmonella in turkeys. Processes that generate significant dust prior to slaughter should be regarded as critical control points for Salmonella.

Increased expression of sialyl-dimeric LeX antigen in liver metastases of human colorectal carcinoma.

We collected a total of 78 tissue specimens, including primary colorectal carcinoma, normal colonic mucosa, and liver metastases of colon carcinoma, to examine whether the extracts of these tissues inhibited the binding of a monoclonal antibody FH6, specific for sialyl-dimeric LeX antigen. The results of inhibition assays demonstrated that: (a) contents of FH6-reactive molecules were greater in carcinoma tissues than in normal colonic mucosa; (b) metastatic foci in livers contained more FH6-reactive molecules than primary tumors; (c) primary tumors from Dukes' stage B1 patients contained less FH6-reactive molecules than primary tumors from Dukes' stage D patients. The inhibitory activity of these tumor tissue extracts against the binding of a monoclonal antibody FH6 to cultured colon carcinoma cells was eliminated by prior treatment of the extracts with sialidase, confirming that the FH6-reactive materials were sialyl-dimeric LeX antigen. Electrophoretic separation of tumor tissue extracts on 3% polyacrylamide gels followed by direct staining with monoclonal antibody FH6 revealed that very high molecular weight glycoproteins, presumably mucins, contained sialyl-dimeric LeX antigen.

Monoclonal antibody against human colonic sulfomucin: immunochemical detection of its binding sites in colonic mucosa, colorectal primary carcinoma, and metastases.

Previous studies using metabolic labeling of fresh colonic mucosa and colorectal carcinoma with [35S]sulfate followed by biochemical analysis demonstrated that the amount of a sulfated high-molecular-weight glycoprotein expressed in primary colorectal carcinoma was lower than that in normal mucosa, and that the amount further decreased in liver metastases. This suggested that this sulfated molecule represented a sulfomucin previously defined by histochemical reactivity with a cationic dye. We have extracted and partially purified this high-molecular-weight sulfated glycoprotein from normal human colonic mucosa. We immunized mice with the partially purified sulfomucin and generated hybridomas. One cloned hybridoma, designated as 91.9H, produced a monoclonal antibody strongly reactive with a component which migrated at an identical position as the metabolically [35S]sulfate-labeled high-molecular-weight glycoprotein after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The reactive molecules appeared to have a polydisperse nature with a molecular weight ranging between 400,000 and 900,000. The [35S]sulfate-labeled high-molecular-weight glycoprotein was bound to Staphylococcus Protein A-agarose coated with this monoclonal antibody but did not bind to unconjugated Protein A-agarose. The immunoprecipitated substance also migrated at an apparent molecular weight range of 400,000 to 900,000. The reactivity of monoclonal antibody 91.9H with the extracts of normal mucosa, colorectal primary carcinoma, and metastasis was compared by dot blot assay on a nitrocellulose membrane. This antibody was more reactive with the extracts of mucosa adjacent to carcinoma tissues than with the carcinoma extracts. Primary tumors showed higher reactivity than metastases in most of the cases. These results strongly suggest that this antibody is specific to colonic sulfomucins or at least to mucins closely related to colonic mucins previously identified by metabolic labeling with [35S]sulfate.

Allergic sensitisation and allergic rhinitis are associated with n-3 polyunsaturated fatty acids in the diet and in red blood cell membranes.

OBJECTIVE: Due to inconsistent results based on dietary intake data, unsaturated fatty acids in red blood cell (RBC) membranes and diet were used to investigate their association with allergic sensitisation and allergic rhinitis. DESIGN: Cross-sectional, population-based study. SETTING: Bavarian Nutrition Survey II (2002-03), Germany. SUBJECTS: A total of 568 adult participants, 325 women and 243 men. METHODS: By means of logistic regression models, the relation of fatty acids to (i) allergic sensitisation as defined by means of specific serum immunoglobulin E analysis (CAPSX1 class > or = 2), and (ii) self-reported allergic rhinitis was examined. RESULTS: A high cell membrane level of eicosapentaenoic acid (EPA, 20:5 n-3) was inversely associated with allergic sensitisation, the adjusted odds ratio (OR) and 95% confidence interval (95% CI) were 0.52 (0.30-0.90) for the highest (vs lowest) quartile. A similar effect was observed for allergic rhinitis with an OR (95% CI) of 0.50 (0.24-1.03; P = 0.027 for trend). A higher dietary intake of alpha-linolenic acid (ALA, 18:3 n-3) was associated with a decreased risk of allergic sensitisation and allergic rhinitis with ORs (95% CIs) of 0.51 (0.28-0.93) and 0.43 (0.20-0.93), respectively, in the highest quartiles. No other dietary or cell membrane unsaturated fatty acid was significantly associated with the outcome variables, nor was the n-6/n-3 ratio. The strongest effects were observed among subjects under the age of 40 y. CONCLUSIONS: In this cross-sectional study among adults, a high content of n-3 fatty acids in RBC membranes (EPA) or in the diet (ALA) is associated with a decreased risk of allergic sensitisation and allergic rhinitis.

The Efficiency of Biofilters at Mitigating Airborne MRSA from a Swine Nursery.

Our prior studies have been in agreement with other researchers in detecting airborne methicillin-resistant Staphylococcus aureus (MRSA) inside and downwind of a swine housing facility. MRSA emitted in the exhaust air of swine facilities creates a potential risk of transmission of these organisms to people in the general area of these facilities as well as to other animals. This study investigated a possible means of reducing those risks. We investigated the efficiency of biofilters to remove MRSA from the exhaust air of a swine building. Two types of biofilter media (hardwood chips and western red cedar shredded bark) were evaluated. Efficiency was measured by assessing both viable MRSA (viable cascade impactor) and dust particles (optical particle courter) in the pre-filtered and post-filtered air of a functioning swine production facility. Our study revealed that hardwood chips were respectively 92% and 88% efficient in removing viable MRSA and total dust particles. Western red cedar was 95% efficient in removing viable MRSA and 86% efficient in removing dust particles. Our findings suggest that biofilters can be used as effective engineering controls to mitigate the transmission of aerosolized MRSA in the exhaust air of enclosed swine housing facilities.

Lesion-induced synaptogenesis in the dentate gyrus of aged rats: II. Demonstration of an impaired degeneration clearing response.

Previously we reported that a delayed onset in the reinnervation of the outer two-thirds of the dentate molecular layer occurred in aged rats after an entorhinal lesion. Several factors associated with formation of new synaptic contacts and removal of degenerative debris may affect the reinnervation process. In this study the appearance and removal of degeneration was analyzed and evaluated with respect to the delayed reinnervation process in aged rats. After a complete lesion of the entorhinal cortex, 85-90% of the input to the outer two-thirds of the ipsilateral molecular layer is lost. Electron-dense and electron-lucent degeneration are present throughout the outer two-thirds of the denervated molecular layer. In both aged and young adult rats, the electron-lucent degeneration disappears by 10 days postlesion. The predominant electron dense degeneration, however, is removed at a different rate by young adult and aged rats. Young adults demonstrate a biphasic degeneration removal process, with almost half of this degeneration rapidly lost by 10 days postlesion, and nearly all by 60 days postlesion. Aged animals in contrast, have lost only 16% of the dense degeneration at 10 days postlesion, with about 30% of the degeneration remaining at 60 days postlesion. The impaired removal of the degeneration from the denervated zone appears to be reciprocally related to the reinnervation response in both age groups and may be related to the normal astrocyte hypertrophy and elevated corticosteroid levels in aged rats.

Lesion-induced synaptogenesis in the dentate gyrus of aged rats: I. Loss and reacquisition of normal synaptic density.

Quantitative electron microscopy was used to examine the ability of aged (2-year-old) and young adult (90-day-old) rats to replace those synapses lost (85-90%) in the outer two-thirds of the molecular layer of the dentate gyrus after a complete unilateral lesion of the entorhinal cortex. In aged rats the synaptic density is significantly lower than that of young adults at 10 days postlesion. Synaptic replacement begins between 2 and 4 days postlesion in young adults, whereas there is a delay until after 10 days postlesion in aged rats. Once synapse replacement begins in aged rats, the rate of synapse reappearance is about equal that of young adults. Thus the initial 10 days postlesion appears critical to growth of responding afferents and reformation of synaptic contacts. Analysis of synapses in terms of noncomplex and complex synaptic types shows that the noncomplex type accounts for the significant synaptic density difference between the two age groups. Replacement of complex synapses is nearly indistinguishable between age groups and is complete by 60 days postlesion. In contrast the initial replacement rate of noncomplex synapses in aged rats is much slower than young adults, though the control synaptic density is achieved by the end of the time course.

Metastatic behavior and cell surface properties of HT-29 human colon carcinoma variant cells selected for their differential expression of sialyl-dimeric Le(x)-antigen.

Immunochemical studies of human colorectal carcinoma with various monoclonal antibodies against Le(X)-related carbohydrate antigens previously revealed that the amount of sialyl-dimeric Le(X) antigen (NeuAc alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-3Gal beta 1-4(Fuc alpha 1-3)GlcNAc beta 1-R: SLX) associated with metastatic lesions was greater than in the primary tumors. To assess whether an experimental model can be used to study the direct relationship between this carbohydrate antigen and the tumor cell's metastatic behavior, we selected variant cells with increased surface SLX from established human colon carcinoma cell line HT-29. The cells in the upper 5% or lower 5% population in fluorescence intensity after reacting with a monoclonal antibody, FH6, were retrieved separately by a fluorescence-activated cell sorter and propagated. After three- or four-times selection, we obtained stable cell lines with low and high cell surface SLX antigens (HT-29 M1 and HT-29 M2, respectively). Binding of monoclonal antibody FH6 was detected to glycolipids extracted from HT-29 M2 cells but not from HT-29 M1 cells. Glycoprotein components having reactivity with monoclonal antibody FH6 were below the detectable level. HT-29 M2 cells injected intrasplenically into nude mice showed a slightly reduced incidence of metastasis to lung, liver and lymph nodes than did HT-29 M1 cells. Subsequently we found that SLX antigen was not detectable by immunohistochemical examination of these tumor cells grown in nude mice. Re-established cell line from nude mice xenografts expressed SLX antigen in vitro.

Bispyridinamines: a new class of topical antimicrobial agents as inhibitors of dental plaque.

A series of N,N'-polyalkylenebis[4-(substituted-amino)pyridines] has been prepared, and members have been evaluated as potential anti-dental plaque agents. From among the most active members of the series, one compound, N,N'-[1,10-decanediyldi-1(4H)-pyridinyl-4-ylidene]bis(1-octanam ine) dihydrochloride, octenidine, was selected as a candidate for clinical study.

Cytokine and IL-12 receptor mRNA discriminate between different clinical subtypes in multiple sclerosis.

Little is known about the involvement of cytokines in the pathogenesis of primary progressive (PP) multiple sclerosis (MS). We evaluated in this cross-sectional study whether IL-18, IL-12p35, IL-12p40, TNF-alpha, IFN-gamma, IL-10, IL-4, TGF-beta, IL-12Rbeta1, and IL-12Rbeta2 mRNA expression in unstimulated white blood cells showed significant differences between relapsing-remitting (RR), secondary progressive (SP) and PP MS patients, and healthy controls. All clinical subtypes showed unique mRNA expression patterns as compared to the controls. Both RR and SP patients displayed increased levels of IL-12p40, IL-18, and TGF-beta mRNA compared to controls, whereas PP patients showed only increased IL-18 mRNA levels. Both in PP and SP patients, IFN-gamma and IL-10 mRNA were decreased compared to RR patients and controls. PP patients were unique in that they showed decreased IL-12Rbeta1 mRNA. In conclusion, our data show that the assessment of cytokine (receptor) mRNA profiles is useful to discriminate between the different clinical subtypes and suggest that different cytokines are involved in the pathogenesis of PP MS as compared to RR and SP MS.

Lesion-induced transneuronal plasticity in the adult rat hippocampus.

The process of reactive synaptogenesis has been demonstrated in several areas of the central nervous system, including the hippocampal dentate gyrus. After a complete unilateral entorhinal lesion, approximately 85% of the input to the outer two-thirds of the ipsilateral dentate molecular layer is lost. Bilateral fluctuations in synaptic density within non-denervated zones of the dentate molecular layer predict further alterations in neural circuitry at sites located transneuronally to the denervated dentate granule cells. Using quantitative electron microscopy, our study demonstrates a complete cycle of synapse loss and reacquisition within the ipsilateral but not contralateral CA4/hilus region of the hippocampal formation. This area is one of the terminal fields for the dentate granule cell mossy fiber axons. In addition the granule cell mossy fiber axons sprout during the postlesion time course and form a significantly increased number of new mossy fiber terminals within the ipsilateral and contralateral CA4/hilus area. Our results indicate that responses to brain injury may no longer be confined to a local denervated site, but probably include polyneuronal circuitry loops, which may encompass one or more areas of the central nervous system. Previous difficulties in providing a close behavioral or functional correlation to localized structural events may be explained by a more global brain response to an injury.

Ultrastructural localization of phenothiazines and tetracycline: a new histochemical approach.

To provide high resolution information on the subcellular localization of the phenothiazines and tetracycline, we have developed a new histochemical method that circumvents the difficulties inherent in classical electron microscopic tissue preparatory procedures. Specific and reliable localizations of these drugs were accomplished by their rapid precipitation with phosphotungstic acid (PTA) at pH 7. A cell suspension of Ehrlich ascites carcinoma cells was incubated with a given drug (2.5 x 10(-4) M) and then briefly cross-linked with 1% glutaraldehyde at 4 degrees C. After washing, the cells were exposed to 2% PTA (pH 7) to precipitate the drug at its binding sites. Then the samples were rapidly dehydrates in 80% ethylene glycol (4 degrees C) and embedded in the polyester, Vestopal W. This protocol provides a low denaturation, low extraction approach to tissue preparation. Control samples (without drug) demonstrated an amorphous distribution of PTA throughout the cell and no specific dense precipitates. Those cells treated with the phenothiazines (chlorpromazine or fluphenazine) or tetracycline demonstrated very discrete (4-8 nm), electron-dense drug-PTA reaction products associated with different nuclear components as well as several cytoplasmic organelles. These subcellular localizations verify the binding sites reported by the biochemical literature. In addition, several previously unresolvable binding sites are reported. The rationale and limitations of this procedure are presented. This new histochemical methodology may have broad applications in the study of drug distribution, receptors, and drug-induced pathology and toxicity that may provide new information regarding drug action and design.

Combined percutaneous carotid stenting and coronary angioplasty during acute ischemic neurologic and coronary syndromes.

This study demonstrates the feasibility of percutaneous carotid and coronary intervention in patients with unstable neurologic and coronary syndromes. Further study is warranted to determine the safety and long-term results in a large series of patients.