Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

<p>Abstract</p> <p>Background</p> <p>The aim of the present study was to investigate biochemical and oxidative stress responses to experimental <it>F. tularensis </it>infection in European brown hares, an important source of human tularemia infections.</p> <p>Methods</p> <p>For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild <it>Francisella tularensis </it>subsp. <it>holarctica </it>strain (a single dose of 2.6 × 10⁹CFU <it>pro toto</it>).</p> <p>Results</p> <p>Subcutaneous inoculation of a single dose with 2.6 × 10⁹colony forming units of a wild <it>F. tularensis </it>strain <it>pro toto </it>resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation.</p> <p>Conclusions</p> <p>Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied.</p

Grief interventions for people bereaved by suicide: A systematic review

<div><p>Background</p><p>Adaption to the loss of a loved one due to suicide can be complicated by feelings of guilt, shame, responsibility, rejection, and stigmatization. Therefore people bereaved by suicide have an increased risk for developing complicated grief which is related to negative physical and mental disorders and an increased risk for suicidal behavior. Grief interventions are needed for this vulnerable population. The aim of this systematic review was to provide an overview of the current state of evidence concerning the effectiveness of interventions that focus on grief for people bereaved by suicide.</p><p>Methods</p><p>We conducted a systematic literature search using PubMed, Web of Science, and PsycINFO for articles published up until November 2016. Relevant papers were identified and methodological quality was assessed by independent raters. A narrative synthesis was conducted.</p><p>Results</p><p>Seven intervention studies met the inclusion criteria. Two interventions were based on cognitive-behavioral approaches, four consisted of bereavement groups, and one utilized writing therapy. As five of the seven interventions were effective in reducing grief intensity on at least one outcome measure, there is some evidence that they are beneficial. Bereavement groups tend to be effective in lowering the intensity of uncomplicated grief, as are writing interventions in lowering suicide-specific aspects of grief. Cognitive-behavioral programs were helpful for a subpopulation of people who had high levels of suicidal ideation.</p><p>Limitation</p><p>On average, methodological quality was low so the evidence for benefits is not robust. The stability of treatment effects could not be determined as follow-up assessments are rare. Generalizability is limited due to homogeneous enrollments of mainly female, white, middle-aged individuals.</p><p>Conclusions</p><p>People bereaved by suicide are especially vulnerable to developing complicated grief. Therefore, grief therapies should be adapted to and evaluated in this population. Prevention of complicated grief may be successful in populations of high risk individuals.</p></div

Bronchoscopic Brushing from Central Lung CancerNext Generation Sequencing Results are Reliable

The role of bronchoscopic brushing for tumor detection and molecular testing in central lung cancer is unclear. In this study, 50 consecutive subjects with suspected central lung cancer underwent bronchoscopic brushing (31 males, median age 70, 5 never smokers). Histological results were: NSCLC/SCLC/low-grade-NET/granulation tissue in 36/8/2/4 cases. Next generation sequencing (NGS) was feasible in 62% of tumor-positive brush smear samples. In 78% of these cases, NGS displayed identical results compared to histology samples, in 22% NGS from brush smears detected specific mutations, whereas DNA quality from forceps biopsy was insufficient for NGS analysis. Sensitivity, specificity, positive predictive value, negative predictive value of brush smear analysis were 66% (95% confidence interval 50-79), 100% (40-100), 100% (85-100), and 21% (7-46). For the combined analysis of brush smear, brush tip washing and sheath tube content sensitivity was slightly elevated at 69% (53-81). In central lung cancer, bronchoscopic brushing detects tumor cells in about two-third of cases and allows a decision for or against targeted therapy in the majority of tumor-positive cases on the basis of NGS analysis

Methodological quality of included studies.

<p>Methodological quality of included studies.</p

PRISMA flow diagram.

<p>PRISMA flow diagram.</p

Characteristics of included studies.

<p>Characteristics of included studies.</p

Description of grief measures.

<p>Description of grief measures.</p

Summary of the included studies without a comparison group.

<p>Summary of the included studies without a comparison group.</p

Comparison of Two Solid-Phase Extraction (SPE) Methods for the Identification and Quantification of Porcine Retinal Protein Markers by LC-MS/MS

Proper sample preparation protocols represent a critical step for liquid chromatography-mass spectrometry (LC-MS)-based proteomic study designs and influence the speed, performance and automation of high-throughput data acquisition. The main objective of this study was to compare two commercial solid-phase extraction (SPE)-based sample preparation protocols (comprising SOLA&#181;TM HRP SPE spin plates from Thermo Fisher Scientific and ZIPTIP&#174; C18 pipette tips from Merck Millipore) for analytical performance, reproducibility, and analysis speed. The house swine represents a promising animal model for studying human eye diseases including glaucoma and provides excellent requirements for the qualitative and quantitative MS-based comparison in terms of ocular proteomics. In total six technical replicates of two protein fractions [extracted with 0.1% dodecyl-&#223;-maltoside (DDM) or 1% trifluoroacetic acid (TFA)] of porcine retinal tissues were subjected to in-gel trypsin digestion and purified with both SPE-based workflows (N = 3) prior to LC-MS analysis. On average, 550 &#177; 70 proteins (1512 &#177; 199 peptides) and 305 &#177; 48 proteins (806 &#177; 144 peptides) were identified from DDM and TFA protein fractions, respectively, after ZIPTIP&#174; C18 purification, and SOLA&#181;TM workflow resulted in the detection of 513 &#177; 55 proteins (1347 &#177; 180 peptides) and 300 &#177; 33 proteins (722 &#177; 87 peptides), respectively (FDR &lt; 1%). Venn diagram analysis revealed an average overlap of 65 &#177; 2% (DDM fraction) and 69 &#177; 4% (TFA fraction) in protein identifications between both SPE-based methods. Quantitative analysis of 25 glaucoma-related protein markers also showed no significant differences (P &gt; 0.05) regarding protein recovery between both SPE methods. However, only glaucoma-associated marker MECP2 showed a significant (P = 0.02) higher abundance in ZIPTIP&#174;-purified replicates in comparison to SOLA&#181;TM-treated study samples. Nevertheless, this result was not confirmed in the verification experiment using in-gel trypsin digestion of recombinant MECP2 (P = 0.24). In conclusion, both SPE-based purification methods worked equally well in terms of analytical performance and reproducibility, whereas the analysis speed and the semi-automation of the SOLA&#181;TM spin plates workflow is much more convenient in comparison to the ZIPTIP&#174; C18 method