Rapid Identification of Phospholipase A2 Transcripts from Snake Venoms

Phospholipase A2 (PLA2) is a major component in snake venoms and it is found in many different isoforms. To identify transcripts encoding different PLA2 isoforms, we developed a simple, rapid procedure. Total RNA was extracted from the venoms of three cottonmouth snakes and two diamondback rattlesnakes, and further reverse-transcribed into complementary DNA (cDNA). Using one pair of cottonmouth PLA2-specific primers and Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique, we identified 27 unique full-length PLA2 transcripts, including nine sequences identical to the previously documented ones in the database and one novel GIII-like PLA2. Two common transcripts respectively encoding Asp49 and Lys49 PLA2 isoforms were identified in all three cottonmouth venoms that contain more PLA2 transcripts than diamondback rattlesnake venoms. The placement of cloned PLA2 transcripts in snake venom PLA2s was further discussed by phylogenetic analysis. The procedure developed in this study paves the way for accelerated acquisition of transcriptome data on any other venom toxin families. The results obtained are crucial for insight into the structure and function of PLA2 isoforms for scientific and potential therapeutic purposes

CONHECIMENTO SOBRE A IMPORTÂNCIA DO DESCARTE CORRETO DE MEDICAMENTOS POR ESTUDANTES DE ENSINO MÉDIO EM PORTO ALEGRE-RS: PROJETO EDUCATIVO DE INTERAÇÃO UNIVERSIDADE-COMUNIDADE

Disposal of medicines is a theme in evolution concerning environmental education, mainly considering the negative effects from disposal in wrong places, like household waste or wastewater system. The present manuscript was developed from a project that works with environment education and health, focusing expired and unused medicines and the importance of their correct disposal, involving high school students in the city of Porto Alegre, Rio Grande do Sul. From the exchange of experiences mediated by the presentation and discussion of educational material, 209 high school students, aged between 15 and 19 years old, from October 2018 to January 2020, participated of a descriptive cross-sectional research. The aim was to evaluate the knowledge of students concerning the correct disposal of expired and unused drugs, through the application of a questionnaire. By descriptive analysis of collected information, it was observed that 82% of the students improperly discard expired and unused medicines, with 49% discarding in dry garbage, 24% in organic waste, 6% in the toilet and 3% in the sink. Of those who dispose correctly (18%), 9% of the students return the medicines to the pharmacy and another 9% to the public health centers. Regarding the reduced number of correct disposal actions, environmental education actions that lead to awareness of the problem that irregular disposal can generate, as well as the transformation of this current scenario, are imperative. It is clear the need for strategies to improve the knowledge about the correct disposal of medicines, especially with the young public, which can disseminate the importance of this action, in favor of the environment and society.O descarte de medicamentos constitui temática em contínua evolução em aspectos educativos ambientais, em especial pelas consequências negativas já conhecidas a partir da disposição deste tipo de resíduo em lixo comum ou em rede de esgoto. O presente artigo origina-se de projeto de extensão que trabalha com educação ambiental e saúde, no foco medicamentos vencidos e em desuso e a importância do seu descarte correto, junto ao ambiente escolar de ensino médio no município de Porto Alegre, Rio Grande do Sul. A partir da troca de experiências mediada pela apresentação e discussão de material educativo, 209 alunos do ensino médio, com idade entre 15 e 19 anos, no período de outubro de 2018 a janeiro de 2020, participaram de pesquisa transversal descritiva. O objetivo foi avaliar os conhecimentos prévios dos alunos sobre o descarte correto de medicamentos vencidos e em desuso (sobras), por meio da aplicação de questionário. Pela análise descritiva dos dados coletados observou-se que 82% dos alunos descartam de forma inadequada os medicamentos vencidos e suas sobras, sendo que 49% descartam em lixo seco, 24% em lixo orgânico, 6% em vaso sanitário e 3% em pia. Daqueles que descartam de modo correto (18%), 9% dos alunos devolvem os medicamentos à farmácia e outros 9% às Unidades de Saúde. Em relação ao quantitativo reduzido de ações corretas de descarte, torna-se imperativo ações de educação ambiental que levem à conscientização da problemática que o descarte irregular pode gerar, bem como à transformação deste cenário. O estudo mostrou a necessidade de estratégias para melhoria no âmbito do conhecimento sobre descarte correto de medicamentos, em especial junto ao público jovem, que pode disseminar a importância desta ação, em prol do meio ambiente e da sociedade

Intravenous human umbilical cord-derived mesenchymal stromal cell administration in models of moderate and severe intracerebral hemorrhage

Intracerebral hemorrhage (ICH) is as a life-threatening condition that can occur in young adults, often causing long-term disability. Recent preclinical data suggests mesenchymal stromal cell (MSC)-based therapies as promising options to minimize brain damage after ICH. However, therapeutic evidence and mechanistic insights are still limited, particularly when compared to other disorders such as ischemic stroke. Herein, we employed a model of collagenase-induced ICH in young adult rats to investigate the potential therapeutic effects of an intravenous injection of human umbilical cord Wharton's jelly-derived MSCs (hUC-MSCs). Two doses of collagenase were used to cause moderate or severe hemorrhages. Magnetic resonance imaging showed that animals treated with hUC-MSCs after moderate ICH had smaller residual hematoma volumes than vehicle-treated rats, whereas the cell therapy failed to decrease the hematoma volume in animals with a severe ICH. Functional assessments (rotarod and elevated body swing tests) were performed for up to 21 days after ICH. Enduring neurological impairments were seen only in animals subjected to severe ICH, but the cell therapy did not induce statistically significant improvements in the functional recovery. The biodistribution of Technetium-99m-labeled hUC-MSCs was also evaluated, showing that most cells were found in organs such as the spleen and lungs 24 h after transplantation. Nevertheless, it was possible to detect a weak signal in the brain, which was higher in the ipsilateral hemisphere of rats subjected to a severe ICH. These data indicate that hUC-MSCs have moderately beneficial effects in cases of less severe brain hemorrhages in rats by decreasing the residual hematoma volume, and that optimization of the therapy is still necessary

Evaluation of predictive factors for i-CLARAS (intraoperative complications in laparoscopic renal and adrenal surgery): a multicentre international retrospective cohort study

The laparoscopic approach represents the standard of treatment for renal and adrenal diseases, and its use is increasing even outside referral centres. Although most procedures are routinely performed, intraoperative complications do not occur, and the rate and predictive factors of these complications have not been established. The aim of this study was to evaluate the incidence and type of intraoperative complications and to identify predictive factors in patients undergoing laparoscopic renal and adrenal surgery. This was a cohort, multicentre, international retrospective study. Patients who underwent laparoscopic renal and adrenal surgeries between April 2017 and March 2022 were included in the study. Bivariate analysis was performed using contingency tables and the χ2 test for independent samples to compare qualitative variables and the T test and Mood test for continuous variables. Multivariate analysis was performed using a logistic regression model to obtain adjusted odds ratios. A total of 2374 patients were included in the study. Intraoperative complications were reported for 8.09% of patients who underwent renal surgery, with the most common complications reported being hollow viscus and vascular complications, and for 6.75% of patients who underwent adrenal surgery, with the most common complication reported being parenchymatous viscous complications. Multivariate analysis revealed that both adrenal and renal surgery radiological preoperative factors, such as invasive features during adrenalectomy and the RENAL score during nephrectomy, are predictive factors of intraoperative complications. In contrast to existing data, surgeon experience was not associated with a reduction in the incidence of perioperative complications

The MMS22L-TONSL Complex Mediates Recovery from Replication Stress and Homologous Recombination

Genome integrity is jeopardized each time DNA replication forks stall or collapse. Here we report the identification of a complex composed of MMS22L (C6ORF167) and TONSL (NFKBIL2) that participates in the recovery from replication stress. MMS22L and TONSL are homologous to yeast Mms22 and plant Tonsoku/Brushy1, respectively. MMS22L-TONSL accumulates at regions of ssDNA associated with distressed replication forks or at processed DNA breaks, and its depletion results in high levels of endogenous DNA double-strand breaks caused by an inability to complete DNA synthesis after replication fork collapse. Moreover, cells depleted of MMS22L are highly sensitive to camptothecin, a topoisomerase I poison that impairs DNA replication progression. Finally, MMS22L and TONSL are necessary for the efficient formation of RAD51 foci after DNA damage, and their depletion impairs homologous recombination. These results indicate that MMS22L and TONSL are genome caretakers that stimulate the recombination-dependent repair of stalled or collapsed replication forks

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Dry Needling as an Alternative Physical Therapy Treatment for Patients With Cervicogenic Headaches: A Systematic Review

Background: There has been an increase in prevalence in cervicogenic headaches (CGH) which is commonly accompanied by neck pain, decreased cervical range of motion (ROM), and other symptoms related to neck movement. Purpose: The objective of this systematic review was to compare manipulation/mobilization versus dry needling (DN) to determine which treatment is most effective in improving pain intensity, ROM, and frequency in individuals with CGH. Methods: Current literature was retrieved from a search using Pubmed and EBSCO (Medline) databases. The following keywords were used to guide searches: “cervicogenic” “headache” “neck pain” “dry needling” “manipulation” “mobilization” “manual therapy”. After filtering, 10 studies met inclusion and exclusion criteria such as treatment via manipulation/mobilization or dry needling, diagnosis of CGH, no other treatments or headache diagnoses were the focus of the study, or outcome measures not aligned with our purpose. All studies were appraised by 2 raters using the PEDro scale, and PRISMA guidelines were followed. Results: Of the 1,406 identified studies, 10 randomized control trials were eligible for review. Only 7 studies met our threshold for quality on the Pedro scale (≥ 5). Two studies of good quality and one of fair quality suggested that DN was an effective treatment; two studies of good quality suggested that manipulation was significantly better than placebo and mobilization; one good quality study suggested that mobilizations worked significantly better than placebo but had a small effect size; and one good quality study suggested that manipulation combined with DN was significantly effective as treatment. Positive treatment benefits for DN were evident immediately after treatment and persisted for up to 6 months, whereas manipulation was evident immediately and for up to 3 months due to the length of the study. The analysis of the data revealed that DN and manipulations significantly improved headache frequency, intensity, and cervical range of motion. However, more research is necessary to establish if a single treatment is sufficient or whether a mix of treatments is more effective. Conclusion: This systematic review found both dry needling and manual therapy techniques as effective treatment methods when treating individuals with cervicogenic headaches. However, due to limited research directly comparing the two treatment methods no treatment is found to be superior than the other for improving pain intensity, cervical ROM, and frequency in patients with cervicogenic headaches

Rapid Identification of Phospholipase A₂ Transcripts from Snake Venoms

Phospholipase A2 (PLA2) is a major component in snake venoms and it is found in many different isoforms. To identify transcripts encoding different PLA2 isoforms, we developed a simple, rapid procedure. Total RNA was extracted from the venoms of three cottonmouth snakes and two diamondback rattlesnakes, and further reverse-transcribed into complementary DNA (cDNA). Using one pair of cottonmouth PLA2-specific primers and Reverse Transcription Polymerase Chain Reaction (RT-PCR) technique, we identified 27 unique full-length PLA2 transcripts, including nine sequences identical to the previously documented ones in the database and one novel GIII-like PLA2. Two common transcripts respectively encoding Asp49 and Lys49 PLA2 isoforms were identified in all three cottonmouth venoms that contain more PLA2 transcripts than diamondback rattlesnake venoms. The placement of cloned PLA2 transcripts in snake venom PLA2s was further discussed by phylogenetic analysis. The procedure developed in this study paves the way for accelerated acquisition of transcriptome data on any other venom toxin families. The results obtained are crucial for insight into the structure and function of PLA2 isoforms for scientific and potential therapeutic purposes

Microemulsions for topical delivery of 8-methoxsalen

8-Methoxsalen (8-MOP) and related furocumarins have been extensively used for the treatment of hyperproliferative skin diseases in association with long-wavelength UVA light. In order to develop alternative formulations for the topical administration of 8-MOP, microemulsions were evaluated as delivery vehicles. Six microemulsion formulations were prepared using water, isopropyl myristate (IPM) and Tween(R) 80. Span(R) 80: 1,2-Octanediol (3:1:1.2 w/w). The microemulsions were characterized using conductimetric and dynamic light scattering analyses. The ability of the systems to deliver 8-MOP into and through the skin was evaluated in vitro using newborn pig-skin. The in vitro permeation data showed that the novel microemulsions increased the 8-MOP total penetration through the skin by order of 1.9-4.5, as compared with IPM. In general, the accumulation of 8-MOP into the skin was increased by a factor of 1.5-4.5 by the microemulsion systems with respect to their total amount of drug delivered across the skin. These results suggest that the studied microemulsion systems may be appropriate vehicles for the topical delivery of 8-MOP. (C) 2000 Elsevier Science B.V. All rights reserved