Infection of Gammarus duebeni populations by two vertically transmitted microsporidia; parasite detection and discrimination by PCR–RFLP

We screened a population of the brackish water crustacean Gammarus duebeni from the Isle of Cumbrae for the presence of vertically transmitted microsporidia. We compared 2 screening techniques; light microscopy and PCR-based detection using generic 16S rDNA microsporidian primers. Fifty percent of females from this population tested positive for vertically transmitted microsporidia. The PCR screen was 100% efficient in comparison with existing LM based screening. In addition, the PCR screen produced bands of 2 sizes suggesting that more than 1 species of microsporidian was present. Sequencing revealed 2 distinct species of vertically transmitted microsporidia; 33% of females were infected with the feminizer Nosema granulosis and 17% were infected with a new species which we provisionally designate Microsporidium sp. On the basis of sequence information, we developed a discriminatory PCR–RFLP test based on MspI and HaeIII digests. This screen allows rapid detection and discrimination of vertically transmitted microsporidia in natural field populations. We applied the PCR–RFLP screen to a second G. duebeni population from the Isle of Man. This population also hosted these 2 parasite species. In total 45% of females harboured N. granulosis and 10% harboured Microsporidium sp. No dual-infected individuals were found in either population. The occurrence of 2 vertically transmitted parasites within a population has implications for our understanding of parasite–host relationships in the field and we discuss factors affecting the dynamics of parasite–parasite competition and coexistence

Fiber depolymerization

Depolymerization is, by definition, a crucial process in the reversible assembly of various biopolymers. It may also be an important factor in the pathology of sickle cell disease. If sickle hemoglobin fibers fail to depolymerize fully during passage through the lungs then they will reintroduce aggregates into the systemic circulation and eliminate or shorten the protective delay (nucleation) time for the subsequent growth of fibers. We study how depolymerization depends on the rates of end- and side-depolymerization, kend and kside, which are, respectively, the rates at which fiber length is lost at each end and the rate at which new breaks appear per unit fiber length. We present both an analytic mean field theory and supporting simulations showing that the characteristic fiber depolymerization time View the MathML source depends on both rates, but not on the fiber length L, in a large intermediate regime 1 much less-than ksideL2/kend much less-than (L/d)2, with d the fiber diameter. We present new experimental data which confirms that both mechanisms are important and shows how the rate of side depolymerization depends strongly on the concentration of CO, acting as a proxy for oxygen. Our theory remains rather general and could be applied to the depolymerization of an entire class of linear aggregates, not just sickle hemoglobin fibers

Estimation of lung growth using computed tomography

Anatomical studies suggest that normal lungs grow by rapid alveolar addition until about 2 yrs of age followed by a gradual increase in alveolar dimensions. The aim of this study was to examine the hypothesis that normal lung growth can be monitored by computed tomography (CT). Therefore, the gas volume per gram of lung tissue was estimated from measurements of lung density obtained from CT scans performed on children throughout the growth period. CT scans were performed on 17 males and 18 females, ranging in age from 15 days-17.6 yrs. CT-measured lung weight was correlated with predicted post mortem values and CT measured gas volume with predicted values of functional residual capacity. The median value for lung expansion was 1.86 mL x g(-1) at 15 days, decreased to 0.79 mL x g(-1) by 2 yrs and then increased steadily to 5.07 mL x g(-1) at 17 yrs. Computed tomography scans can be used to estimate lung weight, gas volume and expansion of normal lungs during the growth period. The increase in the lung expansion after the age of 2 yrs suggests progressive alveolar expansion with increasing lung volume

Activation of neutrophils within pulmonary microvessels of rabbits exposed to cigarette smoke.

Previous studies have shown that polymorphonuclear leukocytes (PMN) are delayed in the pulmonary capillaries by the presence of cigarette smoke. To determine if the PMN delayed by smoking are activated, we estimated the in vivo expression of CD11/CD18 and L-selectin on the surface of PMN in lungs and peripheral blood of rabbits because these molecules are known to be upregulated and downregulated, respectively, on the surface of activated PMN. New Zealand white rabbits (3.5 +/- 0.1 kg) were exposed to either air (n = 5) or cigarette smoke (n = 5), and we used an established protocol to measure pulmonary vascular blood flow, volume, and red blood cell (RBC) transit time in the left lung. The right lungs were then fixed in 0.025% glutaraldehyde and stored in liquid nitrogen. Ultrathin sections were immuno-labeled with either the anti-CD18 monoclonal antibody 60.3 or the anti-L-selectin antibody Dreg-200, followed by a secondary antibody conjugated to 10 nm colloidal gold. The target antigens were quantified by counting the number of gold particles per micron (G/microns) of PMN surface membrane. The data show that smoke exposure had no effect on pulmonary blood flow, volume, or RBC transit time. However, it increased the expression of CD11/CD18 on intravascular PMN in the upper region of the lung (control, 7.4 +/- 1.3 G/microns; smoke-exposed, 13.2 +/- 3.3 G/microns; P < 0.05) and decreased the expression of L-selectin on intravascular PMN in both the lower (control, 5.5 +/- 2.0 G/microns; smoke-exposed, 2.6 +/- 1.5 G/microns; P = 0.05) and the upper (control, 6.8 +/- 1.4 G/microns; smoke-exposed, 2.6 +/- 1.2 G/microns; P < 0.05) regions

Near-Surface Interface Detection for Coal Mining Applications Using Bispectral Features and GPR

The use of ground penetrating radar (GPR) for detecting the presence of near-surface interfaces is a scenario of special interest to the underground coal mining industry. The problem is difficult to solve in practice because the radar echo from the near-surface interface is often dominated by unwanted components such as antenna crosstalk and ringing, ground-bounce effects, clutter, and severe attenuation. These nuisance components are also highly sensitive to subtle variations in ground conditions, rendering the application of standard signal pre-processing techniques such as background subtraction largely ineffective in the unsupervised case. As a solution to this detection problem, we develop a novel pattern recognition-based algorithm which utilizes a neural network to classify features derived from the bispectrum of 1D early time radar data. The binary classifier is used to decide between two key cases, namely whether an interface is within, for example, 5 cm of the surface or not. This go/no-go detection capability is highly valuable for underground coal mining operations, such as longwall mining, where the need to leave a remnant coal section is essential for geological stability. The classifier was trained and tested using real GPR data with ground truth measurements. The real data was acquired from a testbed with coal-clay, coal-shale and shale-clay interfaces, which represents a test mine site. We show that, unlike traditional second order correlation based methods such as matched filtering which can fail even in known conditions, the new method reliably allows the detection of interfaces using GPR to be applied in the near-surface region. In this work, we are not addressing the problem of depth estimation, rather confining ourselves to detecting an interface within a particular depth range

T cells with high PD-1 expression are associated with lower HIV-specific immune responses despite long-term antiretroviral therapy

Objective: We evaluated frequencies of T cells with high PD-1 expression (PD-1HI) before and after long-term effective antiretroviral therapy (ART), and determined if frequencies on-ART correlated positively with measures of HIV persistence and negatively with HIV-specific responses.Methods:We enrolled individuals who started ART during chronic infection and had durable suppression of viremia for at least 4 years (N=99). We assessed PD-1HI T-cell frequencies at timepoints pre-ART and on-ART using flow cytometry, and evaluated how frequencies on-ART are associated with measures of HIV persistence, HIV-specific immune responses, and immune activation levels.Results:Pre-ART, PD-1HI CD4+ T cells correlated positively with viremia and negatively with CD4+ T-cell count. At year 1 on-ART, %PD-1HI CD4+ T cells decreased but then remained stable at 4 and 6-15 years on-ART, whereas %PD-1HI CD8+ T cells on-ART remained similar to pre-ART. PD-1HI CD4+ T cells correlated positively with HIV DNA pre-ART and on-ART, and with CD4+ T-cell activation on-ART. PD-1HI CD4+ T cells negatively correlated with HIV Gag-specific and Env-specific T-cell responses but not with CMV-specific or EBV-specific responses. PD-1HI CD8+ T cells trended towards a negative correlation with responses to Gag and Env, but not to CMV and EBV.Conclusion:PD-1HI T cells persist in blood despite prolonged suppression on ART, correlate with HIV DNA levels, and are associated with lower HIV-specific T-cell responses but not CMV-specific or EBV-specific responses, suggesting that these cells are HIV-specific. The findings support evaluating PD-1 blockade strategies for their effect on HIV persistence and HIV-specific immunity

Cumulative antiretroviral exposure measured in hair is not associated with measures of HIV persistence or inflammation among individuals on suppressive ART

Data on the relationship of antiretroviral exposure to measures of human immunodeficiency virus (HIV) persistence are limited. To address this gap, multiple viral, immunologic, and pharmacologic measures were analyzed from individuals with sustained virologic suppression on therapy (median 7 years) in the AIDS Clinical Trials Group A5321 cohort. Among 110 participants on tenofovir-(TFV)-disoproxil-fumarate (TDF)/emtricitabine (FTC)-containing regimens, we found no significant correlation between hair concentrations of individual antiretrovirals (ARVs) in the regimen and measures of HIV persistence (plasma HIV-1 RNA by single copy assay, cell-associated-DNA, cell-associated RNA) or soluble markers of inflammation. These findings suggest that higher systemic ARV exposure may not impact HIV persistence or inflammation