An Emerging Approach for Parallel Quantification of Intracellular Protozoan Parasites and Host Cell Characterization Using TissueFAXS Cytometry

Characterization of host-pathogen interactions is a fundamental approach in microbiological and immunological oriented disciplines. It is commonly accepted that host cells start to change their phenotype after engulfing pathogens. Techniques such as real time PCR or ELISA were used to characterize the genes encoding proteins that are associated either with pathogen elimination or immune escape mechanisms. Most of such studies were performed in vitro using primary host cells or cell lines. Consequently, the data generated with such approaches reflect the global RNA expression or protein amount recovered from all cells in culture. This is justified when all host cells harbor an equal amount of pathogens under experimental conditions. However, the uptake of pathogens by phagocytic cells is not synchronized. Consequently, there are host cells incorporating different amounts of pathogens that might result in distinct pathogen-induced protein biosynthesis. Therefore, we established a technique able to detect and quantify the number of pathogens in the corresponding host cells using immunofluorescence-based high throughput analysis. Paired with multicolor staining of molecules of interest it is now possible to analyze the infection profile of host cell populations and the corresponding phenotype of the host cells as a result of parasite load.     

Leaf phenology mediates provenance differences in herbivore populations on valley oaks in a common garden

1. Plants from different populations often display a variation in herbivore resistance. However, it is rarely understood what plant traits mediate such differences. 2. It was tested how leaf phenology affects herbivore populations in a 15‐year‐old common garden of valley oaks (Quercus lobata Née) with different populations and maternal parents from throughout the Q. lobata range. 3. The abundance of leaf miners (Stigmella sp. Shrank) and leaf phenology of oaks in the common garden was measured. 4. Leaf miner abundance varied among provenance locations (population), but not among maternal parents within populations. Leaf phenology varied by provenance location and maternal parent, and trees that leafed out earlier accrued higher leaf‐miner abundance. Path analysis indicated that leaf phenology was the likely driver of provenance and parental differences in resistance to leaf miners. 5. Understanding population differences is particularly important when considering transport of genotypes for ornamental or restoration purposes. The present study suggests that similarity in leaf phenology may be one factor that could be used to find genotypes with a similar herbivore resistance to local genotypes.     

The influence of stomach distention on feeding in the nudibranch mollusk Melibe leonina

Although research on satiation has revealed much about the effect of sensory inputs on motivational state, we have yet to fully understand exactly how satiating signals influence the neural circuits underlying specific behaviors. One organism that is well suited for addressing this question is the nudibranch Melibe leonina, because its feeding activity is easily quantified, it has translucent skin that makes the stomach easy to observe, and it has large, identifiable neurons that are very suitable for subsequent analysis of the neural correlates of satiation. In this study our goal was to document the time course of satiation in Melibe, and determine if stomach distention contributes to satiation. When exposed to brine shrimp (Artemia), Melibe immediately commenced stereotypic oral hood movements to capture prey, and continued to do so for approximately five hours. Individuals eventually stopped, despite the continued presence of food, and the slowing and eventual termination of oral hood closures was correlated with distension of the stomach caused by the ingested Artemia. We obtained further evidence that stomach distension is one of the underlying causes of satiation by injecting artificial non-nutritive food into the stomach, and by cutting open part of the stomach wall to prevent it from filling and distending. The first treatment caused satiation to occur more rapidly, while the second treatment delayed satiation. Both results demonstrate that in Melibe stomach distention has a major impact on the motivation to feed. These findings provide the framework for subsequent studies designed to determine precisely how stomach distention influences feeding circuits.     

Serum Levels of Soluble CD26/Dipeptidyl Peptidase-IV in Type 2 Diabetes Mellitus and Its Association with Metabolic Syndrome and Therapy with Antidiabetic Agents in Malaysian Subjects

BackgroundA soluble form of CD26/dipeptidyl peptidase-IV (sCD26/DPP-IV) induces DPP-IV enzymatic activity that degrades incretin. We investigated fasting serum levels of sCD26/DPP-IV and active glucagon-like peptide-1 (GLP-1) in Malaysian patients with type 2 diabetes mellitus (T2DM) with and without metabolic syndrome (MetS), as well as the associations between sCD26/DPP-IV levels, MetS, and antidiabetic therapy.MethodsWe assessed sCD26/DPP-IV levels, active GLP-1 levels, body mass index (BMI), glucose, insulin, A1c, glucose homeostasis indices, and lipid profiles in 549 Malaysian subjects (including 257 T2DM patients with MetS, 57 T2DM patients without MetS, 71 non-diabetics with MetS, and 164 control subjects without diabetes or metabolic syndrome).ResultsFasting serum levels of sCD26/DPP-IV were significantly higher in T2DM patients with and without MetS than in normal subjects. Likewise, sCD26/DPP-IV levels were significantly higher in patients with T2DM and MetS than in non-diabetic patients with MetS. However, active GLP-1 levels were significantly lower in T2DM patients both with and without MetS than in normal subjects. In T2DM subjects, sCD26/DPP-IV levels were associated with significantly higher A1c levels, but were significantly lower in patients using monotherapy with metformin. In addition, no significant differences in sCD26/DPP-IV levels were found between diabetic subjects with and without MetS. Furthermore, sCD26/DPP-IV levels were negatively correlated with active GLP-1 levels in T2DM patients both with and without MetS. In normal subjects, sCD26/DPP-IV levels were associated with increased BMI, cholesterol, and LDL-cholesterol (LDL-c) levels.ConclusionSerum sCD26/DPP-IV levels increased in T2DM subjects with and without MetS. Active GLP-1 levels decreased in T2DM patients both with and without MetS. In addition, sCD26/DPP-IV levels were associated with Alc levels and negatively correlated with active GLP-1 levels. Moreover, metformin monotherapy was associated with reduced sCD26/DPP-IV levels. In normal subjects, sCD26/DPP-IV levels were associated with increased BMI, cholesterol, and LDL-c.     

The ABC’s of Suicide Risk Assessment: Applying a Tripartite Approach to Individual Evaluations

There is considerable need for accurate suicide risk assessment for clinical, screening, and research purposes. This study applied the tripartite affect-behavior-cognition theory, the suicidal barometer model, classical test theory, and item response theory (IRT), to develop a brief self-report measure of suicide risk that is theoretically-grounded, reliable and valid. An initial survey (n = 359) employed an iterative process to an item pool, resulting in the six-item Suicidal Affect-Behavior-Cognition Scale (SABCS). Three additional studies tested the SABCS and a highly endorsed comparison measure. Studies included two online surveys (Ns = 1007, and 713), and one prospective clinical survey (n = 72; Time 2, n = 54). Factor analyses demonstrated SABCS construct validity through unidimensionality. Internal reliability was high (α = .86-.93, split-half = .90-.94)). The scale was predictive of future suicidal behaviors and suicidality (r = .68, .73, respectively), showed convergent validity, and the SABCS-4 demonstrated clinically relevant sensitivity to change. IRT analyses revealed the SABCS captured more information than the comparison measure, and better defined participants at low, moderate, and high risk. The SABCS is the first suicide risk measure to demonstrate no differential item functioning by sex, age, or ethnicity. In all comparisons, the SABCS showed incremental improvements over a highly endorsed scale through stronger predictive ability, reliability, and other properties. The SABCS is in the public domain, with this publication, and is suitable for clinical evaluations, public screening, and research.