Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection.

The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures for monitoring virus circulation and reduce the spreading of SARS-CoV-2. Here, we described quickness and cheaper strategies of direct RT-qPCR (in the absence of RNA isolation) and compared the results to those obtained using standard RNA isolation procedure. The tests varied using pure, diluted samples, combined with Proteinase K (PK) or Lysis Buffer. Our findings showed consistently that PK pre-treated samples in the absence of RNA extraction procedures presents similar results to those obtained by standard RNA isolation procedures. On average, 16 samples extracted with the MagMAX™ CORE Kit, take around 2 h, costing an average of USD 5, the pre-treatment of samples using PK, on the other hand, would cut the value to less than USD 0.30 and reduce the time of procedure in more than 1 ½ hours. The present study suggests the use of PK treatment instead of RNA isolation in order to reduce costs and time in processing samples for molecular diagnosis of SARS-CoV-2.

The auditory P200 is both increased and reduced in schizophrenia? A meta-analytic dissociation of the effect for standard and target stimuli in the oddball task.

OBJECTIVE: Conflicting reports of P200 amplitude and latency in schizophrenia have suggested that this component is increased, reduced or does not differ from healthy subjects. A systematic review and meta-analysis were undertaken to accurately describe P200 deficits in auditory oddball tasks in schizophrenia. METHODS: A systematic search identified 20 studies which were meta-analyzed. Effect size (ES) estimates were obtained: P200 amplitude and latency for target and standard tones at midline electrodes. RESULTS: The ES obtained for amplitude (Cz) for standard and target stimuli indicate significant effects in opposite directions: standard stimuli elicit smaller P200 in patients (d = -0.36; 95% CI [-0.26, -0.08]); target stimuli elicit larger P200 in patients (d = 0.48; 95% CI [0.16, 0.82]). A similar effect occurs for latency at Cz, which is shorter for standards (d = -0.32; 95% CI [-0.54, -0.10]) and longer for targets (d = 0.42; 95% CI [0.23, 0.62]). Meta-regression analyses revealed that samples with more males show larger ES for amplitude of target stimuli, while the amount of medication was negatively associated with the ES for the latency of standards. CONCLUSIONS: The results obtained suggest that claims of reduced or augmented P200 in schizophrenia based on the sole examination of standard or target stimuli fail to consider the stimulus effect. SIGNIFICANCE: Quantification of effects for standard and target stimuli is a required first step to understand the nature of P200 deficits in schizophrenia.

Can muscle fatty acid signature be used to distinguish diets during the marine trophic phase of sea lamprey (Petromyzon marinus, L.)?

Characterization of muscle and liver fatty acid profiles, determination of liver lipogenic and lipolytic activities and estimation of liver fatty elongases and desaturases activities of sea lamprey were realized at the beginning of the spawning migration. The muscle fatty acid profile was consistent with the location of capture, and revealed that animals captured far upstream from the river mouth presented the lowest C18:1ω9 levels and the highest relative proportions of C20:4ω6, C20:5ω3 (EPA), C22:5ω3 (DPA) and C22:6ω3 (DHA). These results suggest: (i) the vital importance of the conservation of C20:4ω6 as a precursor of eicosanoids; (ii) the retention of EPA, DPA and DHA for metabolic energy for reproduction; and (iii) the utilization of C18:1ω9 for metabolic fuel use in the beginning of the spawning period. Hepatic lipolysis and lipogenesis revealed significant differences which could, eventually, result from the diet during the parasitic phase of sea lamprey life cycle. Present results revealed that the muscle act as a fat depot site which explains the few significant correlations observed for fatty acids between muscle and liver. Muscle neutral lipids fatty acid signature at the beginning of the spawning migration can be used to distinguish differences in the diet of sea lampreys during the marine trophic phase of their life cycle.

E-cadherin immunoexpression patterns in the characterisation of gastric carcinoma histotypes.

AIMS: E-cadherin, the main epithelial intercellular adhesion molecule, is abnormally expressed in many cancer types, including gastric carcinoma, which is the second leading cause of cancer death worldwide. The aim of this study was to contribute to the characterisation of gastric carcinoma histotypes based on a new approach to E-cadherin immunoexpression. METHODS: 97 gastric tumour samples obtained from the files of the Hospital of Cancer/Cancer Institute of Ceará, Brazil, were histologically analysed and classified as intestinal (n=40), diffuse (n=34), mixed (n=16) or unclassified (n=7) carcinomas. Immunohistochemistry was performed on the tissue microarray sections. Scores were applied according to the system of Jawhari: 0, no staining; 1, cytoplasmic staining; 2, cytoplasmic and membranous staining in the same case; 3, normal membranous immunoexpression; abnormal patterns: scores 0, 1 and 2; normal pattern: score 3. Jawhari scores were then evaluated utilising another approach: the absence of membranous expression scores (0 and 1) versus the presence of membranous expression (scores 2 and 3). RESULTS: A significant association between membranous expression of E-cadherin and the intestinal histotype (36/40 (90%), and 28/41 (68%) for other histotypes) was found, while diffuse carcinomas were related to the absence of membranous expression. A very strong and peculiar relationship was observed between cytoplasm-exclusive E-cadherin expression (score 1) and the diffuse component of mixed tumours (11/16 (69%)). CONCLUSIONS: E-cadherin immunoexpression patterns help us to characterise gastric carcinoma histotypes. The presence or absence of membranous staining is the most valuable criterion in evaluating E-cadherin expression. Mixed tumours show a characteristic E-cadherin cytoplasmic expression in gastric carcinomas.

Accumulation and biological cycling of heavy metal in four salt marsh species, from Tagus estuary (Portugal).

Pools of Zn, Cu, Cd and Co in leaf, stem and root tissues of Sarcocornia fruticosa, Sarcocornia perennis, Halimione portulacoides and Spartina maritima were analyzed on a bimonthly basis, in a Tagus estuary salt marsh. All the major concentrations were found in the root tissues, being the concentrations in the aboveground organs neglectable for sediment budget proposes, as seen by the low root-aboveground translocation. Metal annual accumulation, root turnovers and cycling coefficients were also assessed. S. maritima showed the higher root turnovers and cycling coefficients for most of the analyzed metals, making this a phytostabilizer specie. By contrast the low root turnover, cycling coefficient and low root necromass generation makes S. perennis the most suitable specie for phytoremediation processes. Although the high amounts of metal return to the sediments, due to root senescence, salt marshes can still be considered sinks of heavy metals, cycling heavy metals mostly between sediment and root.

Effect of novel A2A adenosine receptor agonist ATL 313 on Clostridium difficile toxin A-induced murine ileal enteritis.

Clostridium difficile is a spore-forming, anaerobic, gram-positive bacillus that releases two main virulence factors: toxins A and B. Toxin A plays an important pathogenic role in antibiotic-induced diarrhea and pseudomembranous colitis, a condition characterized by intense mucosal inflammation and secretion. Agonist activity at A2A adenosine receptors attenuates inflammation and damage in many tissues. This study evaluated the effects of a new selective A2A adenosine receptor agonist (ATL 313) on toxin A-induced injury in murine ileal loops. ATL 313 (0.5 to 5 nM) and/or the A2A adenosine receptor antagonist (ZM241385; 5 nM) or phosphate-buffered saline (PBS) were injected into ileal loops immediately prior to challenge with toxin A (1 to 10 microg/loop) or PBS. Intestinal fluid volume/length and weight/length ratios were calculated 3 h later. Ileal tissues were collected for the measurement of myeloperoxidase, adenosine deaminase activity, tumor necrosis factor alpha (TNF-alpha) production, histopathology, and detection of cell death by the TUNEL (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling) method. Toxin A significantly increased volume/length and weight/length ratios in a dose-dependent fashion. ATL 313 treatment significantly (P < 0.05) reduced toxin A-induced secretion and edema, prevented mucosal disruption, and neutrophil infiltration as measured by myeloperoxidase activity. ATL 313 also reduced the toxin A-induced TNF-alpha production and adenosine deaminase activity and prevented toxin A-induced cell death. These protective effects of ATL 313 were reversed by ZM241385. In conclusion, the A2A adenosine receptor agonist, ATL 313, reduces tissue injury and inflammation in mice with toxin A-induced enteritis. The finding of increased ileal adenosine deaminase activity following the administration of toxin A is new and might contribute to the pathogenesis of the toxin A-induced enteritis by deaminating endogenous adenosine.