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To overcome the multifactorial complexity associated with the analysis and interpretation of the capillary electrophoresis results of forensic mixture samples, probabilistic genotyping methods have been developed and implemented as software, based on either qualitative or quantitative models. The former considers the electropherograms' qualitative information (detected alleles), whilst the latter also takes into account the associated quantitative information (height of allele peaks). Both models then quantify the genetic evidence through the computation of a likelihood ratio (LR), comparing the probabilities of the observations given two alternative and mutually exclusive hypotheses. In this study, the results obtained through the qualitative software LRmix Studio (v.2.1.3), and the quantitative ones: STRmix™ (v.2.7) and EuroForMix (v.3.4.0), were compared considering real casework samples. A set of 156 irreversibly anonymized sample pairs (GeneMapper files), obtained under the scope of former cases of the Portuguese Scientific Police Laboratory, Judiciary Police (LPC-PJ), were independently analyzed using each software. Sample pairs were composed by (i) a mixture profile with either two or three estimated contributors, and (ii) a single contributor profile associated. In most cases, information on 21 short tandem repeat (STR) autosomal markers were considered, and the majority of the single-source samples could not be a priori excluded as belonging to a contributor to the paired mixture sample. This inter-software analysis shows the differences between the probative values obtained through different qualitative and quantitative tools, for the same input samples. LR values computed in this work by quantitative tools showed to be generally higher than those obtained by the qualitative. Although the differences between the LR values computed by both quantitative software showed to be much smaller, STRmix™ generated LRs are generally higher than those from EuroForMix. As expected, mixtures with three estimated contributors showed generally lower LR values than those obtained for mixtures with two estimated contributors. Different software products are based on different approaches and mathematical or statistical models, which necessarily result in the computation of different LR values. The understanding by the forensic experts of the models and their differences among available software is therefore crucial. The better the expert understands the methodology, the better he/she will be able to support and/or explain the results in court or any other area of scrutiny., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Arsenopyrite (FeAsS), the most common arsenic-bearing mineral, is usually found associated with pyrite (FeS 2 ) in gold mining tailings. This work examined the galvanic effect of FeS 2 on As release from FeAsS oxidation in circumneutral media under oxygen-depleted and oxygen-saturated conditions. The oxidation experiments were conducted with a flow-through reactor in the absence of FeS 2 particles and in the presence of different contents of this sulfide. The results indicated that the permanent, physical contact between FeAsS and FeS 2 particles causes an increase in the accumulated As release, mainly under O 2 -saturated conditions. At 30% wt. FeS 2 , the increases relatively to FeS 2 -free conditions were 82% and 117% in O 2 -depleted and O 2 -saturated solutions, respectively. At steady-state, the As release rates increased from (4.9 ± 0.5)× 10 -4 µmol m -2 s -1 (0% wt. FeS 2 ) to (1.1-1.9)× 10 -3 µmol m -2 s -1 (5-30% wt. FeS 2 ) under O 2 -saturated conditions. Analysis of FeAsS samples after oxidation revealed oxidized particles partially or entirely covered by precipitates with different sizes, shapes and compositions (e.g., As-S-bearing ferrihydrite, elemental sulfur, and As-O phases). A fine (3-4 nm thick) amorphous layer of S-As-bearing ferric oxy-hydroxide was also identified on oxidized FeAsS, with Fe(III) and As(III) species., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Periaqueductal gray (PAG) is a midbrain region that projects to areas controlling behavioral and autonomic outputs and is involved in the behavioral and physiological components of defense reactions. Since Raphe Pallidus (RPa) is a medial medullary region comprising sympathetic premotor neurons governing heart function, it is worth considering the PAG-RPa path. We assessed: i) whether PAG projects to RPa; ii) the amplitude of cardiac responses evoked from PAG; iii) whether cardiovascular responses evoked from PAG rely on RPa. Experiments conducted in Wistar rats (±300 g) were approved by Ethics Committee CEUA-UFG (092/18). Firstly, (n = 3), monosynaptic retrograde tracer Retrobeads was injected into RPa; PAG slices were analyzed. Other two groups (n = 6 each) were anesthetized with urethane (1.4 g/kg) and chloralose (120 mg/kg) and underwent craniotomy, tracheostomy, catheterization of femoral artery and vein and of cardiac left ventricle. In one group, we injected the GABA A receptor antagonist, bicuculline methiodide (BMI - 40 pmol/100 nL) into lateral/dorsolateral PAG. Another group was injected (100 nL) with the GABA A receptor agonist muscimol (20 mM) into RPa, 20 min before BMI into PAG. The results were: i) retrogradely labelled neurons were found in PAG; ii) PAG activation by BMI caused positive chronotropism and inotropism, which were accompanied by afterload increases; iii) RPa inhibition with Muscimol reduced heart rate, arterial and ventricular pressures; iv) the subsequent PAG activation still increased arterial pressure, cardiac chronotropy and inotropy, but these responses were significantly attenuated. In conclusion, PAG activation increases cardiac chronotropy and inotropy, and these responses seem to rely on a direct pathway reaching ventromedial medullary RPa neurons., Competing Interests: Declaration of competing interest No conflicts of interest, financial or otherwise, are declared by the authors., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The lack of tissue selectivity of anticancer drugs generates intense collateral and adverse effects of cancer patients, making the incorporation of vitamins or micronutrients into the diet of individuals to reduce side or adverse effects of antineoplastics. The study aimed to evaluate the effects of retinol palmitate (RP) on the toxicogenic damages induced by cyclophosphamide (CPA), doxorubicin (DOX) and its association with the AC protocol (CPA + DOX), in Sarcoma 180 (S-180) tumor cell line, using the micronuclei test with a block of cytokinesis (CBMN); and in non-tumor cells derived from Mus musculus using the comet assay. The results suggest that CPA, DOX and AC protocol induced significant toxicogenic damages (P < 0.05) on the S-180 cells by induction of micronuclei, cytoplasmic bridges, nuclear buds, apoptosis, and cell necrosis, proving their antitumor effects, and significant damage (P < 0.001) to the genetic material of peripheral blood cells of healthy mice, proving the genotoxic potential of these drugs. However, RP modulated the toxicogenic effects of antineoplastic tested both in the CBMN test (P < 0.05), at the concentrations of 1, 10 and 100 IU/mL; as in the comet assay (P < 0.001) at the concentration of 100 IU/kg for the index and frequency of genotoxic damage. The accumulated results suggest that RP reduced the action of antineoplastics in non-tumor cells as well as the cytotoxic, mutagenic, and cell death in neoplastic cells., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Endothelium-derived prostacyclin and nitric oxide elevate platelet cyclic nucleotide levels and maintain quiescence. We previously demonstrated that a synergistic relationship exists between cyclic nucleotides and P2Y 12 receptor inhibition. A number of clinically approved drug classes can modulate cyclic nucleotide tone in platelets including activators of NO-sensitive guanylyl cyclase (GC) and phosphodiesterase (PDE) inhibitors. However, the doses required to inhibit platelets produce numerous side effects including headache., Objective: We investigated using GC-activators in combination with P2Y 12 receptor antagonists as a way to selectively amplify the anti-thrombotic effect of both drugs., Methods: In vitro light transmission aggregation and platelet adhesion under flow were performed on washed platelets and platelet rich plasma. Aggregation in whole blood and a ferric chloride-induced arterial thrombosis model were also performed., Results: The GC-activator BAY-70 potentiated the action of the P2Y 12 receptor inhibitor prasugrel active metabolite in aggregation and adhesion studies and was associated with raised intra-platelet cyclic nucleotide levels. Furthermore, mice administered sub-maximal doses of the GC activator cinaciguat together with the PDE inhibitor dipyridamole and prasugrel, showed significant inhibition of ex vivo platelet aggregation and significantly reduced in vivo arterial thrombosis in response to injury without alteration in basal carotid artery blood flow., Conclusions: Using in vitro, ex vivo, and in vivo functional studies, we show that low dose GC activators synergize with P2Y 12 inhibition to produce powerful anti-platelet effects without altering blood flow. Therefore, modulation of intra-platelet cyclic nucleotide levels alongside P2Y 12 inhibition can provide a strong, focused anti-thrombotic regimen while minimizing vasodilator side effects., (© 2020 The Authors. Journal of Thrombosis and Haemostasis published by Wiley Periodicals LLC on behalf of International Society on Thrombosis and Haemostasis.)

Objectives: Anticholinergic drugs have several side effects, and they have been associated with adverse outcomes, particularly in older patients. The aim of this study was to analyze anticholinergic burden and its relationship to delirium and mortality in older acutely ill medical patients., Design: Cohort study., Setting and Participants: Patients 65 years of age and older who were admitted to an Internal Medicine ward between August 1 and December 31, 2016., Methods: Anticholinergic drug use, outpatient and inpatient, was assessed using the Anticholinergic Cognitive Burden Scale (ACB). Prevalent delirium was diagnosed by the Short Confusion Assessment Method (Short-CAM)., Results: Of the 198 patients, 28.3% developed delirium. Mortality rate was 13.6% in-hospital and 45.6% at 12 months. In multivariate analysis, outpatient ACB was associated with delirium, with an odds ratio (OR) of 1.65 [95% confidence interval (CI) 1.09-2.51]. Those with delirium had longer hospital stays (median 13 vs 8 days; P = .01), received more drugs (median 18 vs 15; P = .02), and presented a higher inpatient ACB (mean 3.9 vs 3.1; P = .034). No increased risk was found for in-hospital or 12-month mortality with drug use, ACB, or delirium., Discussion: In the population studied, we found an association between anticholinergic burden as measured by the ACB and the presence of delirium, but not with mortality at 12 months. A very high 12-month mortality rate might have been an obstacle for association recognition., Conclusions and Implications: Clinician awareness of possible drug side effects, especially in older populations, is crucial. As part of medication reconciliation at the time of hospitalization, ACB of prehospitalization medications should be routinely calculated by inpatient pharmacy services and made available to medical teams., (Copyright © 2019 AMDA – The Society for Post-Acute and Long-Term Care Medicine. Published by Elsevier Inc. All rights reserved.)

Forensic laboratories frequently receive biological samples from crime scene investigations with limited DNA in terms of quantity and quality. Hair shafts are a common type of evidence constituting an ideal sample for DNA testing. Nevertheless, the majority of these hairs are rootless. In an attempt to facilitate the analyses of such samples, new DNA quantification (InnoQuant ® HY) and amplification (InnoTyper ® 21) kits were developed, using mobile elements as molecular markers. The main advantage of the InnoTyper ® 21 kit is its ability to amplify short DNA fragments (60-125 bp), overcoing the difficulties in the amplification of samples with low DNA quantity and/or quality. Therefore, the aim of the present work is to evaluate the efficiency of these two kits in rootless hair samples. In this study, the InnoQuant ® HY kit revealed that nearly 94% of the samples presented moderate to high DNA degradation, allowing the selection of the most suitable amplification protocol to be applied. The InnoTyper ® 21 kit revealed that nearly 95% of the samples presented amplification peaks in its genetic profiles (GP). However, 60% of these samples did not present a good enough quality to be reported according to the Best Practice Manuals (BPMs), guidelines and documents published by the DNA Working Group of European Network of Forensic Science Institutes (ENFSI). Moreover, in approximately 23% of the amplified samples, GPs with enough quality to be reported were retrieved, 10% of which exhibiting a complete GP. The use of this kit allowed obtaining results which could not be produced by the kits routinely used at forensic laboratories. These results demonstrated that InnoQuant ® HY and InnoTyper ® 21 kits are reliable alternatives for the analysis of nuclear DNA in challenging samples such as rootless hair shafts., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Anisakiasis is an emerging marine food-borne zoonosis resulting from the accidental ingestion of Anisakis larvae, through the consumption of raw or undercooked infected seafood products. The first case of human gastric hyperinfection by Anisakis simplex with an unusual and severe presentation, occurring in a Portuguese woman, is described in this article. Over 140 anisakid larvae were removed by gastroscopy. Massive infection is uncommon in areas where the consumption of raw fish is not part of the traditional diet, as is the case in Portugal. The increased consumption of raw seafood products is considered a health determinant in the rise in cases of anisakiasis. However, clinicians should be aware of the emergence of these infections, not only because of the new dietary habits of the population, but also because of the high prevalence of Anisakis larvae in the different fish species usually consumed by the population, collected on the Portuguese coast., (Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The central and peripheral renin-angiotensin systems are known for playing a key role in cardiovascular control. In the present study, we evaluated the hemodynamic effects produced by nanoinjections of angiotensin II (Ang II) or angiotensin-(1-7) [Ang-(1-7)] into the rostral ventrolateral medulla (RVLM) of adult male normotensive (Wistar-WT) and spontaneously hypertensive rats (SHR). Animals were anesthetized (urethane 1.2g/kg) and instrumented for recording blood pressure (BP), heart rate (HR) and blood flow (BF) in the femoral, renal or mesenteric arteries. Afterwards, rats were positioned in a stereotaxic and prepared for nanoinjections (100 nl) of saline (NaCl 0.9%), Ang-(1-7) (40 ng) or Ang II (40 ng) into the RVLM. The vascular resistance (VR) was calculated by ΔMAP/ΔBF ratio. In WT, Ang-(1-7) or Ang II caused equipotent pressor effects that were not accompanied by changes in vascular resistance. However, MAP changes were greater in SHR. This strain also showed a concomitant increase in relative vascular resistance (ΔVR/VRbaseline) of renal (0.31 ± 0.07 and 0.3 ± 0.07 vs. 0.02 ± 0.01; Ang-(1-7), Ang II and Saline, respectively) and mesenteric beds (0.3 ± 0.06 and 0.33 ± 0.04 vs. 0.05 ± 0.02; Ang-(1-7), Ang II and saline, respectively). We conclude that Ang II and Ang-(1-7) at the RVLM control the vascular resistance of renal and mesenteric beds during hypertension., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The strategy of antiangiogenic drugs is based on inhibiting formation of new blood vessels as alternative to limit cancer progression. In this work, we investigated the antitumor and antiangiogenic potential of eight thalidomide derivatives. Most of the molecules was not cytotoxic but 2a, 2d and 3d revealed weak antiproliferative activity on HL-60, Sarcoma 180 (S180) and normal peripheral blood mononuclear cells. Thalidomide, 2a and 2b were able to inhibit tumor growth (53.5%, 67.9% and 67.4%, respectively) in S180-bearing mice and presented moderate and reversible toxicity on liver, kidneys and spleens. Both analogs (2a and 2b) inhibited cell migration of endothelial (HUVEC) and melanoma cells (MDA/MB-435) at 50μg/mL. Immunohistochemistry labeling assays with CD-31 (PECAM-1) antibody showed microvascular density (MVD) was significantly reduced in thalidomide, 2a and 2b groups (30±4.9, 64.6±1.8 and 46.5±19.5%, respectively) (p<0.05). Neovascularization evaluated by Chorioallantoic Membrane Assay (CAM) with compounds 2a and 2b showed reduction of vessels' number (12. 9±2.3 and 14.8±3.3%), neovascularization area (13.1±1.7 and 14.3±1.7%) and total length of vessels (9.2±1.5 and 9.9±1.9%). On the other hand, thalidomide did not alter vascularization parameters. Consequently, addition of thiosemicarbazone pharmacophore group into the phthalimidic ring improved the in vivo antitumor and antiangiogenic potential of the analogs 2a and 2b., (Published by Elsevier Ireland Ltd.)

Casearia sylvestris Swartz (Salicaceae) is a plant commonly widespread in the Americas. It has oxygenated tricyclic bioactive clerodane diterpenes with antimicrobial, antiulcer, larvicidal, chemopreventive, anti-inflammatory, antioxidant and antiproliferative properties. Due to this requirement for the developing of new anticancer drugs, it was initially evaluated the cytotoxic activity of a fraction with Casearins (FC) and its clerodane diterpenes Casearin B (Cas B), D (Cas D), X (Cas X) and Caseargrewiin F (Cas F) isolated from C.sylvestris leaves against 7 tumor cell lines, Sarcoma 180 cells (S180) and on normal peripheral blood mononuclear cells (PBMC). All substances tested showed cytotoxic potential. Cas F and X were the most active compounds. Cell death analyzes with Cas F (0.5 and 1μM) and Cas X (0.7 and 1.5μM) using the HL-60 leukemia line as experimental model showed DNA synthesis and membrane integrity reduction, DNA fragmentation and mitochondrial depolarization, specially after 24h exposure, cell cycle arrest in G0/G1 phase caused by Cas X, activation of the initiator -8/-9 and effector -3/-7 caspases and phosphatidylserine externalization, all biochemical features of apoptosis corroborated by chromatinic condensation, karyorrhexis, cytoplasmic vacuolation and rarefaction and cellular shrinkage, morphological findings specially observed after 12 and 24h of incubation. Therefore, Cas X and F were the most functional molecules with more pronounced lethal and discriminating effects on tumor cells and antiproliferative action predominantly mediated by apoptosis, highlighting clerodane dipertenes as promising lead antineoplastic compounds., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Clerodane diterpenes have demonstrated cytotoxic, antiplasmodial and anti-ulcer properties. In the present work, we determined the cytotoxic effect of casearin L (Cas L), O (Cas O) and X (Cas X) and (-)-hardwickiic acid isolated from Casearia sylvestris leaves, and investigated the underlying mechanisms involved in in vitro cell death induced by Cas X in HL-60 leukemia cells (0.7, 1.5 and 3.0μM). Cytotoxicity tests demonstrated that Cas X was the most active compound studied, showing greater cytotoxic effects against CEM and HL-60 lines (IC(50) of 0.4μM) and human peripheral blood mononuclear cells (PBMC, IC(50) of 1.2μM). After 24h exposure, Cas X caused a decrease in 5-bromo-20-deoxyuridine (BrdU) incorporation (36.6 and 24.5% labeling at 0.7 and 1.5μM, respectively), reduction in viability, and increase in apoptotic and necrotic leukemia cells in a dose-dependent manner evidenced by the trypan blue and AO/EB (acridine orange/ethidium bromide) assays. Moreover, Cas X-treated cells exhibited nuclear fragmentation and cytoplasmic vacuolization depending on the concentration tested. These characteristics of apoptosis or secondary necrosis were confirmed by flow cytometry which revealed DNA fragmentation, phosphatidylserine externalization, activation of the effector caspases 3/7 and mitochondrial depolarization. We then found evidence that Cas X causes cell death via apoptotic pathways, corroborating the potential of casearins as compounds with promising antitumor-related properties., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

Using the epilepsy model obtained by systemic administration of pilocarpine in rats in the present study we investigated the changes caused by seizures on content and species of gangliosides and phospholipids, as well as on cholesterol concentration, glutathione reduced contents, Na(+), K(+)-ATPase activity and lipid peroxidation levels in rat hippocampus. Wistar rats received pilocarpine hydrochloride (400mg/kg, i.p., pilocarpine group), and other group received 0.9% saline (i.p., control group). Results showed that seizures significantly decreased the total content of lipids and glutathione reduced concentration in rat hippocampus. We also observed that seizures significantly reduced the absolute quantity of the major brain gangliosides (GM1, GD1a, GD1b and GT1b) and phospholipids (sphingomyelin, phosphatidylcholine and phosphatidylethanolamine). Our data also showed a decreased Na(+), K(+)-ATPase activity and an increased TBARS levels in hippocampus of seized rats. If confirmed in human beings, these data could suggest that the alteration in lipid composition, Na(+), K(+)-ATPase activity, glutathione reduced content and TBARS levels caused by seizures might contribute to the neurophysiopathology of seizures observed in epileptic patients.