Evaluation of the circulating endothelial cell in deep venous thrombosis : patients and animal model

Orientador: Joyce Maria Anichino-BaizzacchiTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências MédicasResumo: As células endoteliais participam da hemostasia com efeitos pró e antitrombóticos, que podem ser estimulados por uma lesão endotelial. A presença de células endoteliais na circulação pode ser considerada um novo marcador de integridade vascular, como já descrito em várias patologias tais como: doenças cardiovasculares, doenças infecciosas, doenças imunes, transplantes, anemia falciforme. Os objetivos do nosso estudo foram: padronizar a identificação e quantificação das células endoteliais circulantes (CECs) e das células endoteliais progenitoras (CEPs) em um grupo de pacientes com trombose venosa profunda (TVP) ao diagnóstico (aguda, 1ª coleta) e após no mínimo 6 meses (2ª coleta), em um grupo de TVP crônica e em um grupo controle; padronizar um modelo animal de TVP induzida por lesão endotelial para avaliar as CECs e CEPs no sangue periférico e no trombo venoso. O grupo de TVP aguda foi composto por 9 pacientes [F: 7; M: 2; 45 anos (26 - 54 anos)], sendo recrutados 6 indivíduos para uma 2ª coleta [F: 5; M: 1; 47,5 anos (27 - 55 anos)], no grupo de TVP crônica foram incluídos 10 pacientes [F: 6; M: 4; 44,5 anos (28 - 56 anos)] e no grupo controle 11 voluntários [F: 9; M: 2; 29 anos (21 - 52 anos)]. A identificação das CECs e CEPs no sangue periférico foi realizada por citometria de fluxo. No modelo animal, a TVP foi induzida por lesão endotelial com uma solução de FeCl3 a 15%. Após a indução da TVP as CECs e CEPs foram avaliadas no sangue periférico nos tempos: 15 minutos, 30 minutos, 45 minutos, 1 hora, 24 horas, 48 horas e 72 horas. O trombo venoso formado na veia cava inferior (VCI) foi avaliado pela coloração de Verhoff van Gienson e a presença de CECs e CEPs por imunofluorescência. Houve uma diferença estatística no número das CECs (P=0.001, CD31+CD144+CD45dimCD133-; P<0.001, CD31+CD146+CD45dimCD133-; P=0.002, CD31+VEGFR2+CD45dimCD133-) entre os grupos estudados, com um aumento importante nos pacientes com TVP aguda. Os grupos de TVP crônica e TVP 2ª coleta mostraram um aumento significativo de CECs em relação aos controles. Não houve diferença estatística no número das CEPs (CD34+VEGFR2+CD45dimCD133-) entre os grupos estudados, porém observou-se um aumento no grupo de TVP aguda. No modelo animal, a oclusão total da VCI foi verificada entre 15 minutos e 1 hora. Após 24 horas ocorreu uma diminuição progressiva da área do trombo formado [85,4% (24h); 65,4% (48h); 51,3% (72h)], e não foram observadas CECs e CEPs no mesmo. No sangue periférico, no tempo de 48 horas houve um aumento importante de CECs e CEPs quando comparado com os outros tempos estudados (P=0.001, Sca1-VEGFR2+CD45dim; P=0.004, CD34-VEGFR2+CD45dim; P<0.001,Sca1+VEGFR2+CD45dim; P<0.001, CD34+VEGFR2+CD45dim). Os resultados observados em camundongos e humanos sugerem que as CEPs podem ser recrutadas da medula óssea para a circulação, participando do processo de reparo endotelial. O aumento de CECs na fase em que se inicia o processo de recanalização no modelo animal, sugere que apesar da lesão endotelial inicial, as mesmas somente são liberadas quando o fluxo começa a ser restabelecido. Neste estudo concluímos que as CECs podem constituir um marcador de dano vascular.Abstract: Endothelial cells participate in hemostasia and have pro and anti -thrombotic effects which can be stimulated by an endothelial lesion. The presence of endothelial cells in circulation can be considered a novel marker of vascular integrity, as described in several pathologies, such as: cardiovascular disease, infectious disease, immune diseases, transplants, and sickle cell anemia. The aims of our study were to: standardize the identification and quantification of circulating endothelial cells (CECs) and progenitor endothelial cells (EPCs) in a group of patients with Deep Vein Thrombosis (DVT) at diagnosis (acute, 1st collection) and after a minimum of six months (2nd collection), in a group with chronic DVT and in a control group; standardize an animal model with DVT induced by endothelial lesion in order to evaluate CECs and EPCs in peripheral blood and in venous thrombosis. The group of DVT was composed of 9 patients [F: 7; M: 2; 45 years old (y.o.) (26 - 54 y.o.)], of which six individuals were recruited for the 2nd collection [F: 5; M: 1; 47.5 y.o. (27 - 55 y.o.)]; in the chronic DVT group 10 patients were included [F: 6; M: 4; 44.5 y.o. (28 - 56 y.o.)]; and in the control group 11 volunteers were included [F: 9; M: 2; 29 y.o. (21 - 52 y.o.)]. The identification of CECs and EPCs in peripheral blood was carried out by flow cytometry. DVT was induced in the animal model by endothelial lesion using a FeCl3 solution at 15%. After DVT induction, CECs and EPCs were evaluated in peripheral blood at: 15 minutes, 30 minutes, 45 minutes, 1 hour, 24 hours, 48 hours and 72 hours. The venous thrombus formed in the inferior cava vein (ICV) was evaluated using Verhoff van Gienson stain and the presence of CECs and EPCs was evaluated using immunofluorescence. There was a statistical difference in the number of CECs (P=0.001, CD31+CD144+CD45dimCD133-; P<0.001, CD31+CD146+CD45dimCD133-; P=0.002, CD31+VEGFR2+CD45dimCD133-) between the groups studied, with a significant increase in acute DVT patients. The chronic DVT and 2nd collection DVT groups demonstrated a significant increase in CECs in relation to the controls. There was no statistical difference in the number of EPCs (CD34+VEGFR2+CD45dimCD133-) among the groups studied, however an increase in the acute DVT group was observed. In the animal model, total ICV occlusion was observed between 15 minutes and 1 hour. After 24 hours a progressive decrease in the area of the formed thrombus occurred [85.4% (24h); 65.4% (48h); 51.3% (72h)], and no CECs or CEPs were observed. After 48hours, there was a significant increase in CECs and EPCs in peripheral blood when compared to other periods studied (P=0.001, Sca1-VEGFR2+ CD45dim; P=0.004, CD34-VEGFR2+CD45dim; P<0.001, Sca1+VEGFR2+CD45dim; P<0.001, CD34+VEGFR2+CD45dim). The results observed in mice and human beings suggest that EPCs can be recruited to circulation from the bone marrow, participating in a process of endothelial repair. The increase of CECs during the phase when the re-channeling process begins in the animal model, suggests that despite the initial endothelial lesion, CECs are liberated when the flow is reestablished. In the present study we concluded that CECs can represent a vascular damage marker.DoutoradoMedicina ExperimentalDoutor em Ciência

Salvage procedure after sternochondroplasty for pectus excavatum.

Vacuum-Assisted Closure in association to flap reconstruction is a well-established approach for treating complicated wounds. Case presentation: The authors present a case of soft-tissue breakdown with hardware exposure in a patient treated with strenochondroplasty for pectus excavatum. VAC therapy was applied in association to multiple debridement procedures and final flap closure. This was achieved without the need for hardware removal, thus maintaining adequate skeletal fixation. To our knowledge this approach has not yet been described into the literature after sternochondroplasty procedure. Conclusion Aggressive debridement and VAC therapy before final closure may represent a new, conservative method for managing surgical complications after sternochondroplasty procedures

Suplementação em excesso com Ácido Fólico durante o pré-natal: revisão da literatura / Excessive supplementation with Folic Acid during prenatal: literature review

A suplementação com ácido fólico para evitar defeitos no fechamento do tubo neural é recomendada como parte do cuidado pré-natal. As diretrizes recomendam administrar 400µg diários de AF, não podendo esta dose ultrapassar 1000µg diários. No entanto, não é incomum nos depararmos com casos em que a ingestão de AF supera as doses recomendadas, o que constitui uma realidade preocupante, uma vez que os efeitos decorrentes do uso crônico de doses elevadas desta vitamina ainda não são totalmente esclarecidos. Foi realizada uma análise da literatura dos últimos 10 anos, foram selecionados 09 artigos. Alguns dos malefícios que podem estar associados ao excesso de AF no pré-natal incluem aumento do risco de desenvolvimento de asma na infância, bem como infecções de via aérea superior e chiados; exacerbação de deficiência de B12; comprometimento do crescimento fetal; influência no desenvolvimento do cérebro (autismo); reações alérgicas (eczema); promoção do câncer (tumor de mama e adenoma colorretal); aumento resistência à insulina nas crianças; e risco de diabetes gestacional. Esse estudo alerta sobre a importância da adequação do SUS em fornecer comprimidos de AF na dose adequada para gestante, uma vez que os comprimidos ofertados pelo SUS são na dose de 5mg, o que é aproximadamente 10 vezes maior que o recomendado.

Protection of Farms from Wolf Predation: A Field Approach

The livestock sector is facing serious challenges in combatting the increasing predation of domestic livestock. In this scenario, wild carnivores, especially wolves, represent key predators. To allow the coexistence of wild and domestic animals, defense methodologies consisting of multiple integrated antipredator strategies must be tested and implemented based on the geographical man agement context of each farm. This study investigated the potential of a novel antipredator method (PAN, Project Farmers-Nature in Italian) in protecting livestock (goats and horses) from wolves on a farm located in the Sibillini Mountains National Park, over a three-month period (June–September 2022). The PAN field approach involved two phases: (1) interviews with farmers and inspections of how the farm and pasture are structured and (2) monitoring predator abundance using camera traps and transects in order to understand the wildlife habits. Information on predator movement around the grazing area was shared with the farmer, who was actively involved in implementing strategies to protect livestock. The stable presence of one pair of wolves was confirmed in the grazing area, placing grazing livestock at risk. The farmer was advised to strengthen the existing antipredator strategy (herd protection dogs) by introducing two trained Maremma-Abruzzese sheepdog puppies to protect his animals. The implemented actions demonstrate how professional experts can serve as a strategic intermediary between livestock and wildlife conservation in the management of the current conflicts

Pectoralis Muscle Transposition in Association with the Ravitch Procedure in the Management of Severe Pectus Excavatum.

Background: Pectus excavatum (PE) is the most common congenital chest wall deformity. PE is sometimes associated with cardiorespiratory impairment, but is often associated with psychological distress, especially for patients in their teenage years. Surgical repair of pectus deformities has been shown to improve both physical limitations and psychosocial well-being in children. The most common surgical approaches for PE treatment are the modified Ravitch technique and the minimally invasive Nuss technique. A technical modification of the Ravitch procedure, which includes bilateral mobilization and midline transposition of the pectoralis muscle flap, is presented here. Methods: From 2010 to 2016, 12 patients were treated by a modified Ravitch procedure with bilateral mobilization and midline transposition of the pectoralis muscle flap for severe PE. Outcomes, morphological results, and complications were analyzed with respect to this new combined surgical approach. Results: There was a statistically significant difference between pre- and postoperative values (P = 0.0025) of the Haller index at the 18-month follow-up, showing a significant morphological improvement for all treated patients. After surgery, no morbidity and mortality were noted. The mean hospital stay was 7 days, and all patients were discharged without major complications. Conclusion: This technique significantly improved patients’ postoperative morphological outcomes and significantly reduced long-term complications, such as wound dehiscence, skin thinning, and hardware exposure

Comparison of epidemiological profile of leprosy from 2010 to 2020 in the city of Alfenas, state of Minas Gerais, with Brazil

Leprosy is an infectious disease caused by Mycobacterium leprae, a mycobacterium that affects skin cells and peripheral nerves and, when not treated properly, can cause skin lesions with loss of local sensitivity. Its study is still of great importance, as it is a stigmatizing disease, with impacts on public health in emerging countries such as Brazil, and prevalent cities such as the Alfenas city in Minas Gerais. Objective: To compare the epidemiological profile of patients affected by leprosy from 2010 to 2020 in Alfenas city, state of Minas Gerais, with Brazil. Method: This is a transversal and descriptive study with collected data for the period of 2010 to 2020 available in the DATASUS system. Result: The observed profile was characterized by men, aged over 15 years, incomplete Elementary School I and multibacillary type of the disease, however there was divergence in relation to the race/color of individuals affected by leprosy comparing the national scope with the municipal scope of Alfenas. This difference in data is due to the socioeconomic development process in the city of Alfenas since its population composition is similar to that of west Paulista by virtue of the coffee labor in the colonial period. The statistical analysis indicated a varied prevalence coefficients for Brazil and Alfenas, and a dependence between both variables, although relatively weak. Conclusion: With the high prevalence of leprosy cases and its consequences, such as skin lesions, it is of great importance to understand this disease, as well as its epidemiological patterns

The 15-Months Clinical Experience of SARS-CoV-2: A Literature Review of Therapies and Adjuvants

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the virus responsible for the coronavirus disease of 2019 (COVID-19) that emerged in December 2019 in Wuhan, China, and rapidly spread worldwide, with a daily increase in confirmed cases and infection-related deaths. The World Health Organization declared a pandemic on the 11th of March 2020. COVID-19 presents flu-like symptoms that become severe in high-risk medically compromised subjects. The aim of this study was to perform an updated overview of the treatments and adjuvant protocols for COVID-19. Methods: A systematic literature search of databases was performed (MEDLINE PubMed, Google Scholar, UpToDate, Embase, and Web of Science) using the keywords: “COVID-19”, “2019-nCoV”, “coronavirus” and “SARS-CoV-2” (date range: 1 January 2019 to 31st October 2020), focused on clinical features and treatments. Results: The main treatments retrieved were antivirals, antimalarials, convalescent plasma, immunomodulators, corticosteroids, anticoagulants, and mesenchymal stem cells. Most of the described treatments may provide benefits to COVID-19 subjects, but no one protocol has definitively proven its efficacy. Conclusions: While many efforts are being spent worldwide in research aimed at identifying early diagnostic methods and evidence-based effective treatments, mass vaccination is thought to be the best option against this disease in the near future

Rapporto 2007 su consumo e dipendenze da sostanze in Emilia-Romagna.

Report on the state of legal and illegal substances use in the territory of Emilia-Romagna Region.Il report analizza il fenomeno delle dipendenze nel territorio della Regione Emilia-Romagna. La descrizione del fenomeno si sviluppa intorno all\u27analisi degli indicatori individuati dall\u27Osservatorio Europeo delle Dipendenze di Lisbona (OEDT): 1-uso di sostanze nella popolazione generale (questo indicatore va a rilevare i comportamenti nei confronti di alcol e sostanze psicoattive da parte della popolazione generale); 2-prevalenza d\u27uso problematico delle sostanze psicoattive; 3-domanda di trattamento degli utilizzatori di sostanze; 4-mortalit? degli utilizzatori di sostanze; 5-malattie infettive. Altri due importanti indicatori che si stanno sviluppando, e che vengono qui illustrati, sono l\u27analisi delle Schede di Dimissione Ospedaliera (SDO) e gli indicatori relativi alle conseguenza sociali dell\u27uso di droghe (criminalit? droga correlata). Inoltre sono state applicate diverse metodologie standard di stima sia per quantificare la quota parte sconosciuta di utilizzatori di sostanze che non afferiscono ai servizi, sia per identificarne alcune caratteristiche

Guidelines for the use and interpretation of diagnostic methods in adult food allergy

Food allergy has an increasing prevalence in the general population and in Italy concerns 8 % of people with allergies. The spectrum of its clinical manifestations ranges from mild symptoms up to potentially fatal anaphylactic shock. A number of patients can be diagnosed easily by the use of first- and second-level procedures (history, skin tests and allergen specific IgE). Patients with complex presentation, such as multiple sensitizations and pollen-food syndromes, frequently require a third-level approach including molecular diagnostics, which enables the design of a component-resolved sensitization profile for each patient. The use of such techniques involves specialists' and experts' skills on the issue to appropriately meet the diagnostic and therapeutic needs of patients. Particularly, educational programs for allergists on the use and interpretation of molecular diagnostics are needed