Resumen: Los objetivos de esta investigación fueron evaluar la inmunocompetencia frente al virus de la leucosis bovina enzoótica (VLBE), virus de la diarrea viral bovina (VDVB) y herpesvirus bovino 1 (HVB-1) asociada con el suministro de calostro materno (CM) y reemplazador de calostro (RC) en terneras desde el nacimiento, así como valorar el efecto de la transferencia pasiva y la lactoterapia sobre la inmunocompetencia de las terneras desde los 55 hasta los 102 días de edad. Cuarenta terneras fueron asignadas sistemáticamente a uno de dos grupos, como sigue: (1) suministro de ≈ 171 g de IgG en 1,8 L de CM (grupo A1, n=20) y (2) suministro de ≈ 150 g de IgG en 1,5 L de RC (grupo A2, n=20). Luego, se incluyó la aplicación de CM intramuscular a los 55 días de edad para formar dos bloques, como sigue: (1) sin CM intramuscular (grupo N0, n=17), (2) con CM intramuscular (grupo N1, n=18) y cuatro combinaciones entre niveles de A × N (A1N0, n=10; A1N1, n=11; A2N0, n=7 y A2N1, n=7). Muestras de sangre, suero, heces o secreción respiratoria fueron analizadas periódicamente mediante pruebas serológicas o reacción en cadena de la polimerasa desde el nacimiento hasta los 102 días de edad. Los títulos de anticuerpos medidos por el porcentaje de inhibición (PI) frente al VLBE, VDVB y HVB-1 fueron mayores y persistentes en el grupo A1 en comparación con el grupo A2, pero la diferencia entre medias no fue significativa. El PI-seronegativo frente al VDVB se asoció significativamente con la ingesta de RC desde los 60 hasta los 81 días de edad. La vacunación aumentó el número de terneras seropositivas frente al VDVB hasta los 102 días de edad en el grupo A2. La lactoterapia no cambió la tendencia de los niveles de anticuerpos frente al VDVB ni de los otros virus. En conjunto, la lactoterapia con CM intramuscular no afectó la inmunocompetencia frente a los virus, pero en terneras calostradas con CM fue mejor que la proveída con RC.

Resumo: Os objetivos desta pesquisa foram avaliar a imunocompetência contra o vírus da leucose bovina enzoótica (BLV), vírus da diarreia viral bovina (BVDV) ou herpesvírus bovino 1 (BHV-1) associados ao colostro materno (MC) e substituto do colostro (CR) em bezerros desde o nascimento; bem como avaliar a efeito da transferência passiva e da lactoterapia na imunocompetência em bezerros de 55 a 102 dias de idade. Quarenta bezerros foram sistemáticamente designados para um dos dois grupos a seguir: (1) recebeu, via oral, ≈ 171 g de IgG em 1,8 L de colostro materno (CM) (grupo A1, n=20) e (2) recebeu, via oral, ≈ 150 g de IgG em 1,5 L de substituto de colostro (SC) (grupo A2, n=20). Em seguida, foi administrado CM, via intramuscular, aos bezerros com 55 dias de idade para formar dois outros grupos organizados da seguinte forma: (1) sem CM intramuscular (grupo N0, n=17), (2) com CM intramuscular (grupo N1, n=18) e quatro combinações entre os níveis A × N (A1N0, n=10; A1N1, n=11; A2N0, n=7 e A2N1, n=7). Amostras de sangue, soro, fezes ou secreção respiratória foram testadas periódicamente por testes sorológicos ou reação em cadeia da polimerase, desde o nascimento até 102 dias de idade. Os títulos de anticorpos medidos por inibição percentual (PI) contra BLV, BVDV e BHV-1 eram mais altos e persistentes no grupo A1 em comparação com o grupo A2, mas a diferença entre os meios não era significativa. O PI-soronegativo contra BVDV foi significativamente associado ao consumo de SC dos 60 aos 81 dias de idade. A vacinação aumentou o número de bezerros soropositivos BVDV até 102 dias de idade no grupo SC. A colostroterapia não mudou a tendência dos níveis de anticorpos contra BVDV ou os outros vírus. Em geral, a colostroterapia não afetou a imunocompetência contra os vírus, mas o resultado do grupo de bezerros alimentados com CM via oral foi melhor do que aqueles que ingeriram SC.

Abstract: Forty calves were systematically assigned to one of two groups, as follows: (1) supply of ≈ 171 g of IgG in 1.8 L of MC (A1 group A1, n=20) and (2) supply of ≈ 150 g of IgG in 1.5 L of CR (group A2, n=20). Then, the application of intramuscular MC at 55 days of age was included to form two blocks, as follows: (1) without intramuscular MC (group N0, n=17), (2) with intramuscular MC (group N1, n=18) and four combinations between levels of A × N (A1N0, n=10; A1N1, n=11; A2N0, n=7 and A2N1, n=7). Blood, serum, stool or respiratory secretion samples were periodically analyzed by serological tests or polymerase chain reaction from birth to 102 days of age. Antibody titers measured by percent inhibition (PI) against BLV, BVDV, and BHV-1 were higher and more persistent in the group A1 compared to the group A2, but the difference between means was not significant. PI-seronegative BVDV was significantly associated with the intake of CR from 60 to 81 days of age. Vaccination increased the number of BVDV seropositive calves up to 102 days of age in the CR A2. Lactotherapy did not change the trend of antibody levels against BVDV and other viruses. Overall, lactotherapy with intramuscular MC did not affect immunocompetence against viruses, but in calves that ingest MC it was better than the one provided with CR.

ABSTRACT: This study evaluated the vaccine-induced serological response after administering four commercial formulations containing Bovine Viral Diarrhea Virus (BVDV) type-1, BVDV-2, and Bovine Herpesvirus type 1 (BoHV-1) to young heifers with circulating maternal antibody titers. The study also determined the anamnestic response to vaccinations after the complete metabolization of maternal antibodies when the calves reached six months. Eighty-seven Holstein heifers were selected and randomly distributed into four experimental groups based on the vaccine administered. The four vaccine-based experimental groups were as follows: vaccine A (n = 22), vaccine B (n = 22), vaccine C (n = 24), and vaccine D (n = 19), given on the 60th, 90th, and 180th day of life, respectively. Virus-neutralization (VN) tests were performed at baseline (day 60) and 30 days after administering the second (day 120) and third doses (day 210). We analyzed the effect of vaccine group (P ≤ 0.001), day of vaccination (P ≤ 0.001), and group versus vaccine day interaction (P ≤ 0.001) for antibody titers produced against BVDV-1, BVDV-2, and BoHV-1 using the PROC MIXED method (Statistical Analysis System -SAS 9.4). Antibody titers against BVDV-1, BVDV-2, and BoHV-1 were similar at baseline and on day 60 of life. The mean antibody titers were constant and persisted against BVDV-1 in heifers immunized with vaccines A and C. Heifers immunized with vaccine A alone had a similar effect against BVDV-2. Regarding BoHV-1, the antibody titers decreased between days 60 and 210 in groups B, C, and D. The antibody titer for heifers in group A also decreased between days 60 and day 120, and an intense increase in titers was observed on day 210. After being immunized with formulations B, C, and D, the frequency of animals with titers above protective levels for BVDV-1, BVDV-2, and BoHV-1 was very low or null. Indicators of anamnestic response were observed in heifers vaccinated with formulation A only. Therefore, it can be concluded that passive immunity negatively interferes with antibody production after vaccination. ABSTRACT vaccineinduced induced BVDV (BVDV type1, type1 1, type-1 BVDV2, BVDV2 2, 2 BVDV-2 BoHV1 BoHV (BoHV-1 months Eightyseven Eighty seven administered vaccinebased follows n 22, 22 , 22) 24, 24 24) 19, 19 19) 60th th 90th life respectively Virusneutralization neutralization VN (VN 3 120 . 210) P 0.001, 0001 0.001 0 001 BVDV1, BVDV1 BoHV- Statistical SAS 9.4. 94 9.4 9 4 9.4) 6 BVDV- BVDV2. 2. BoHV1, 21 null only Therefore type- (BoHV- 12 000 0.00 00 9. (BoHV 0.0 0.

RESUMO: O objetivo desse estudo foi avaliar a resposta sorológica induzida após vacinação com quatro formulações comerciais contendo o BVDV-1, BVDV-2 e BoHV-1 em novilhas jovens com títulos de anticorpos maternos circulantes, assim como analisar uma possível resposta anamnéstica em vacinações, após a completa metabolização dos anticorpos maternais aos seis meses de idade. Foram selecionadas 87 novilhas Holandesas, distribuídas aleatoriamente em quatro grupos experimentais de acordo com as vacinas: vacina A (n = 22), vacina B (n = 22), vacina C (n = 24) e vacina D (n = 19), aplicadas aos dias 60, 90 e 180 de vida. Testes de vírus-neutralização (VN) foram realizados no momento basal (D60) e 30 dias após a aplicação da 2ª dose (D120) e 3ª dose (D210). Observou-se efeito do grupo (P ≤ 0,001), dia da vacinação (P ≤ 0,001), e interação grupo versus dia (P ≤ 0,001) para os títulos de anticorpos produzidos contra o BVDV-1, BVDV-2 e BoHV-1, por meio do comando PROC MIXED (Statistical Analysis System, versão 9,4). Os títulos de anticorpos contra o BVDV-1, BVDV-2 e BoHV-1 eram semelhantes no momento basal aos 60 dias de vida. Os títulos médios de anticorpos foram constantes e persistentes contra o BVDV-1 nas novilhas imunizadas com as vacinas A e C, porém apenas a vacina A teve este mesmo perfil contra o BVDV-2. Em relação ao BoHV-1, os grupos B, C e D apresentaram queda nos títulos de anticorpos do D60 ao D210, enquanto as novilhas do grupo A apresentaram queda do D60 ao D120, com aumento intenso dos títulos no D210. A frequência de animais com títulos acima dos protetores contra o BVDV-1, BVDV-2 e BoHV-1 foram muito baixos ou nulos após as vacinações nos animais imunizados com as formulações B, C e D. Sugere-se que a imunidade passiva interferiu negativamente na indução de anticorpos pelas vacinas, observando-se indicadores de resposta anamnéstica apenas para as novilhas vacinadas com a formulação A. RESUMO BVDV1, BVDV1 BVDV 1, 1 BVDV2 2 BVDV- BoHV1 BoHV BoHV- circulantes idade 8 Holandesas n 22, 22 , 22) 24 19, 19 19) 9 18 vida vírusneutralização vírus neutralização VN (VN (D60 3 ª D120 (D120 D210 . (D210) Observouse Observou se P 0,001, 0001 0,001 0 001 BoHV1, Statistical System 9,4. 94 9,4 4 9,4) 6 BVDV2. 2. D6 Sugerese Sugere observandose observando (D6 D12 (D12 D21 (D210 000 0,00 00 9, (D D1 (D1 D2 (D21 0,0 (D2 0,

RESUMEN Introducción: La infección aguda por chikungunya produce una infección auto-limitada caracterizada principalmente por fiebre alta, artralgias, artritis, cefalea, mialgias y exantema pruriginoso; una minoría presenta síntomas persistentes que puede evolucionar a la cronicidad. Objetivo: describir las características clínicas y laboratoriales de pacientes adultos ambulatorios con chikungunya en un hospital de referencia durante la epidemia del 2022-2023 en Paraguay. Metodología: estudio descriptivo de corte trasversal de pacientes adultos con chikungunya que consultaron en el Hospital Nacional de Itauguá entre enero-marzo de 2023. Resultados: Se incluyeron 140 pacientes entre 19 y 79 años (edad media: 46,6±13,6), 77,9 % del sexo femenino y 89,3 % del departamento Central. El 72,9 % de los pacientes fueron confirmados por RT-PCR y el resto por la serología o nexo epidemiológico. El 48,6 % tenía alguna comorbilidad, el 65% se encontraba en la fase aguda, y el resto en la fase subaguda (35 %). Los síntomas más frecuentes fueron la poliartralgia (96,4 %); artritis (98,6 %), fiebre (95,7 %), astenia (100 %), mialgias (96,4 %), cefalea (82,9 %), exantema (95 %). Los síntomas gastrointestinales fueron menos frecuentes; náuseas (37 %), vómitos (15,7 %) y diarrea (20 %). Entre los principales hallazgos laboratoriales se encontraron la proteína C reactiva aumentada (45,7 %), leucopenia (23,6 %), eritrosedimentación aumentada (27,9 %), citólisis hepática leve (13,6 %). Todos fueron tratados con paracetamol; el 60,4 % y el 87,8 % de los pacientes en fase aguda y subaguda recibieron AINES, respectivamente. Discusión: La combinación de fiebre alta y artralgia severa fue el rasgo característico de la infección por el virus chikungunya en la epidemia en Paraguay.

ABSTRACT Introduction: Acute infection by chikungunya produces a self-limited infection characterized mainly by high fever, arthralgia, arthritis, headache, myalgia and pruritic rash; a minority presents persistent symptoms that can evolve to chronicity. Objective: to describe the clinical and laboratory characteristics of adult ambulatory patients with chikungunya at a referral hospital during the 2022-2023 epidemic in Paraguay. Method: descriptive cross-sectional study of adult patients with chikungunya infection who consulted at the Hospital National de Itauguá between January-March 2023. Results: 140 patients between 19 and 79 years old (mean age: 46.6 ± 13.6), 77.9% female and 89.3% from the Central department, were included. 72.9% of the patients were confirmed by RT-PCR and the rest by serology or epidemiological nexus. 48.6% had some comorbidity, 65% were in the acute phase, and the rest in the subacute phase (35%). The most frequent symptoms were polyarthralgia (96.4%); arthritis (98.6%), fever (95.7%), fatigue (100%), myalgia (96.4%), headache (82.9%), rash (95%). Gastrointestinal symptoms were less frequent such as; nausea (37%), vomiting (15.7%) and diarrhea (20%). Increased C-reactive protein (45.7%), leukopenia (23.6%), increased erythrocyte sedimentation rate (27.9%), and mild hepatic cytolysis (13.6%) were among the main laboratory findings. All patients were treated with paracetamol; 60.4% and 87.8% of the patients in the acute and subacute phase, respectively, received NSAIDs. Discussion: The combination of high fever and severe arthralgia can be considered the characteristic feature of chikungunya virus infection in this epidemic.

Resumen Introducción: el virus Chikungunya (CHIKV) es un virus ARN que se transmite a través de mosquitos; se asocia a manifestaciones articulares, tanto en fases agudas, como también a síntomas persistentes articulares pasada la infección. Objetivos: describir la prevalencia informada de artritis reumatoidea (AR) en la infección por CHIKV en sus diferentes fases (aguda, subaguda crónica) y la persistencia de artralgias en la fase crónica. Materiales y métodos: revisión bibliográfica, no sistemática, de artículos científicos publicados desde 2001 hasta 2022, en inglés y español, en las siguientes bases de datos: Medline, Cochrane, Lilacs y Scielo. No se hicieron restricciones en base al tipo de estudio. Se revisaron los resúmenes y, en caso de ser necesario, se evaluaron los artículos completos teniendo en cuenta para su selección los trabajos que describían la asociación de CHIKV AR en la infección por CHIKV en sus diferentes fases (aguda, subaguda y crónica) y la persistencia de artralgias en la fase crónica. Para la búsqueda de artículos en los diferentes motores de búsqueda se emplearon las siguientes ecuaciones de búsqueda: “Chikungunya virus” y “arthritis”; “Chikungunya virus” y “persistent arthralgias”. Como criterio de inclusión, se aplicaron los artículos que mencionaban la infección en adultos y describieran la presencia de AR en la infección por CHIKV en sus diferentes fases (aguda, subaguda y crónica) y la persistencia de artralgias en la fase crónica. Luego de la búsqueda inicial, se seleccionaron 26 artículos que se consideraron relevantes para esta revisión. De los 26 artículos seleccionados, siete fueron revisiones bibliográficas no sistémicas, seis estudios preclínicos (tres en ratones, dos de secuenciación viral y uno de inmunidad celular), cuatro estudios corte transversal descriptivos, cuatro longitudinales prospectivos, dos estudios retrospectivos, una revisión sistémica con metaanálisis, una guía de práctica clínica y una serie de casos clínicos. Resultados: la prevalencia informada de pacientes con AR por CHIKV que progresan a una etapa crónica varía de 4,1 a 78,6%. Las rodillas, tobillos, codos, muñecas y articulaciones metacarpofalángicas son las articulaciones más frecuentemente comprometidas en la fase crónica. El mecanismo por el cual el CHIKV se induce la AR crónica aún está en investigación, y es la persistencia del virus en macrófagos y el mimetismo molecular las teorías más aceptadas hasta el momento. Con respecto al desarrollo de AR en pacientes luego de la infección por CHIKV, no hay a la fecha una postura definida. Existe consenso respecto a que la AR es la enfermedad reumática inflamatoria más frecuentemente encontrada en pacientes recuperados de CHIKV. Dos estudios demostraron que entre un 5 y un 36% de los pacientes que fueron evaluados por artralgias post-CHIKV cumplían criterios del American College of Rheumatology (ACR 2010) de AR. Conclusiones: el compromiso musculoesquelético, principalmente las artralgias crónicas son una manifestación frecuente en un gran número de pacientes recuperados de CHIKV. Si bien hay discrepancia entre los diferentes autores, existe una prevalencia aproximada del 30% de síntomas articulares crónicos posteriores a la infección viral. En general, el compromiso es poliarticular, simétrico y afecta principalmente a las articulaciones de la mano y las rodillas. Deben realizarse más investigaciones y estudios para establecer guías de abordaje de los pacientes con compromiso articular y antecedentes de infecciones por arbovirus.

Abstract Introduction: the Chikungunya virus (CHIKV) is an RNA virus that is transmitted through mosquitoes, which is associated with joint manifestations both in acute phases as well as persistent joint symptoms after the infection. Objectives: to describe the reported prevalence of arthritis in CHIKV infection in its different phases: acute, subacute, chronic, and the persistence of arthralgia in the chronic phase. Materials and methods: non-systematic bibliographic review of scientific articles published from 2001 to 2022, in English and Spanish, in the following databases: Medline, Cochrane, Lilacs, Scielo. No restrictions were made based on the type of study. The abstracts were reviewed and, if necessary, the complete articles were evaluated, taking into account for their selection the works that described the association of the CHIKV virus with arthritis in its different phases: acute, subacute, chronic, and the persistence of arthralgia in the chronic phase. To search for articles in the different search engines, the following search equations were used: “Chikungunya virus” and “arthritis”; “Chikungunya virus” and “persistent arthralgias”. Articles that spoke of infection in adults and described the presence of arthritis and chronic arthralgia once the acute and subacute phases, respectively, were applied as inclusion criteria. After the initial search, 26 articles were considered relevant for this review. Of the 26 articles selected, 7 were non-systemic bibliographic reviews, 6 preclinical studies (3 in mice, 2 on viral sequencing and 1 on cellular immunity), 4 descriptive cross-sectional studies, 4 prospective longitudinal, 2 retrospective studies, 1 systemic review with meta-analysis, 1 clinical practice guideline and 1 series of clinical cases. Results: The reported prevalence of patients with CHIKV arthritis progressing to a chronic stage ranges from 4.1 to 78.6%. The knees, ankles, elbows, wrists, and metacarpophalangeal joints are the most frequently involved joints in the chronic phase. The mechanism by which CHIKV induces chronic arthritis remains under investigation, with virus persistence in macrophages and molecular mimicry being the most accepted theories to date. Regarding the development of Rheumatoid Arthritis (RA) in patients after CHIKV infection, there is no defined position to date. There is consensus that RA is the most frequently found inflammatory rheumatic disease in patients recovered from CHIKV. Two studies have shown that between 5% and 36% of patients who were evaluated for post-CHIKV arthralgia met the 2010 ACR criteria for RA. Conclusions: Musculoskeletal compromise, mainly chronic arthralgia, is a frequent manifestation in a large number of patients recovered from CHIKV. Although there is discrepancy between the different authors, there is an approximate prevalence of 30% of chronic joint symptoms after viral infection. The compromise is generally polyarticular, symmetrical and mainly affects joints of the hand and knees. More research and studies must be carried out in order to establish management guidelines for patients with joint involvement and a history of arbovirus infections.

RESUMO JUSTIFICATIVA E OBJETIVOS: Indivíduos após infecções virais permanecem com sintomas persistentes, como a dor e a fadiga. Exercícios físicos têm sido descritos como alternativa promissora para o controle desses sintomas, porém não há revisões sistemáticas que verifquem a eficácia dessa terapêutica e que avaliem a qualidade destes estudos. O objetivo deste estudo foi investigar o efeito de exercícios físicos na dor ou fadiga associados a infecções virais. MÉTODOS: Revisão sistemática registrada na PROSPERO (CRD42021265174). A coleta de dados foi realizada entre julho de 2021 a janeiro de 2022. Foram incluídos ensaios clínicos randomizados que abordaram a prática de exercícios, em indivíduos com idade superior a 18 anos, com diagnóstico de infecção viral associada à presença de dor ou fadiga por mais de três meses. A busca foi realizada nas bases de dados Pubmed, EMBASE, LILACS e Scielo e, a seleção por pares foi realizada no software (rayyan.ai); a análise de risco de viés foi avaliada através da ferramenta Cochrane risk-of-bias tool for randomized trials 2; a certeza da evidência por meio da GRADE; e para a construção da meta-análise, o software Review Manager. RESULTADOS: Foram selecionados 11 ensaios clínicos nas populações com Vírus da Imunodeficiência Adquirida (HIV), Virus Linfotrópico da Célula T Humana (HTLV), Chikungunya e Poliomielite. Tanto para dor quanto para a fadiga, a conjunção de exercícios aeróbicos com treino resistido, com duração de 40 a 60 minutos, de duas a três vezes por semana, foram eficazes e seguros. A qualidade metodológica dos estudos demonstrou em seis estudos alto risco de viés, devido aos domínios: viés devido a desvios das intervenções pretendidas, viés devido à falta de dados de resultado e viés na seleção do resultado relatado; classificado como algumas preocupações em um estudo devido o domínio viés devido a desvios das intervenções pretendidas; e os demais foram avaliados como baixo risco de viés. Na meta-análise foi demonstrado resultado a favor do grupo intervenção sobre a intensidade da dor nos estudos para Chikungunya e em um estudo para HTLV, o que aponta para efeito positivo a favor dos grupos ativos. CONCLUSÃO: Os exercícios físicos no tratamento da fadiga apresentam evidências muito baixas, enquanto para o desfecho dor os exercícios resistidos apresentam moderada evidência. São recursos de baixo risco e custo, com efeitos promissores, que devem ser melhor testados em pessoas após infecções virais. DESTAQUES O treinamento aeróbico combinado com o treinamento de resistência mostra resultados promissores para reduzir a dor e a fadiga nesta população após a infecção viral. O Pilates é um método que reduz significativamente a intensidade da dor após infecções por HTLV-1 e Chikungunya. O exercício pode beneficiar pessoas com sintomas persistentes de dor e fadiga após infecções

ABSTRACT BACKGROUND AND OBJECTIVES: Individuals after viral infections remain with persistent symptoms such as pain and fatigue. Physical exercises have been described as a promising alternative for the control of these symptoms, but there are no systematic reviews that verify the effectiveness of this therapy and that assess the quality of these studies. The aim of this study was to investigate the effect of physical exercise on pain or fatigue associated with viral infections. METHODS: Systematic review registered with PROSPERO (CRD42021265174). Data collection was carried out between July 2021 and January 2022. Randomized clinical trials that addressed the practice of exercises, in individuals over 18 years of age, diagnosed with viral infection associated with the presence of pain or fatigue for more than 3 months were included. The search was carried out in the Pubmed, EMBASE, LILACS and Scielo databases, and the paired selection was carried out in the software (rayyan.ai); risk of bias analysis was assessed using the Cochrane risk-of-bias tool for randomized trials 2; certainty of evidence through GRADE; and for the construction of the meta-analysis, the Review Manager software. RESULTS: Eleven clinical trials were selected in populations with acquired immunodeficiency virus (HIV), human T-cell lymphotropic virus (HTLV), chikungunya and poliomyelitis. For both pain and fatigue, the combination of aerobic exercise with resistance training, lasting 40 to 60 minutes, two to three times a week, was effective and safe. The methodological quality of the studies showed a high risk of bias in six studies due to the following domains: bias due to deviations from the intended interventions, bias due to lack of outcome data and bias in the selection of the reported outcome; rated as some concerns in one study due to the domain bias due to deviations from intended interventions; and the others were assessed as low risk of bias. The meta-analysis showed a result in favor of the intervention group on pain intensity in the studies for Chikungunya and in a study for HTLV, which points to a positive effect in favor of the active groups. CONCLUSION: Exercises for the treatment of fatigue have very low evidence, while resistance exercises have moderate evidence for pain outcome. These are low-risk, low-cost resources with promising effects that should be better tested in people after viral infections.

ABSTRACT Background: the SARS-CoV-2 virus is responsible for the second pandemic of the 21st century. Since its appearance in China at the end of 2019, it has been associated with pneumonia and considered to be just another respiratory virus. However, during its global spread, it shows its ability to damage other organs with clinical manifestations never before described for other respiratory viruses. Objective: to describe the scientific evidence that supports the direct extra-pulmonary damage produced by the SARS-CoV-2 virus in late stages of infection, which supports its biphasic nature and different from other respiratory viruses. Methods: a search of the articles was carried out in the MEDLINE databases accessed from PubMed, SciELO and LILACS. Articles published in prepress repositories such as medRxiv, BioRxiv were also taken into account. Using the Mendeley reference manager and search manager, duplicates and those that did not meet the objective of the study were eliminated, selecting 63 articles for the present review. Results: the evidence suggests that SARS-CoV-2 has a tropism not only limited to the respiratory tract. The clinical progression of COVID-19 presents a biphasic course, with flu-like manifestations in the first phase and post-acute and persistent episodes in the late phase, caused by direct damage to the central nervous, cardiovascular, endocrine and renal systems. Conclusions: SARS-CoV-2 infection should not be considered only as an acute infection limited to the respiratory tract.

RESUMEN Fundamento: el virus SARS-CoV-2 es responsable de la segunda pandemia del siglo XXI. Desde su aparición en China a finales de 2019, se asocia a neumonía y considera como un virus respiratorio más. Sin embargo, durante su diseminación global demuestra su capacidad para producir daño a otros órganos con manifestaciones clínicas nunca antes descritas para otros virus respiratorios. Objetivo: describir la evidencia científica que respalde el daño extrapulmonar directo producido por el virus SARS-CoV-2 en etapas tardías de la infección, que apoyan su naturaleza bifásica y distinta frente a otros virus respiratorios. Métodos: se realizó una búsqueda de artículos referente al tema en las bases de datos MEDLINE accedido desde PubMed, SciELO y LILACS. También se tuvieron en cuenta artículos publicados en los repositorios de preimpresión como medRxiv, BioRxiv. Mediante el gestor de búsqueda y administrador de referencias Mendeley, se eliminaron los duplicados y aquellos que no se ajustaban al objetivo del estudio, se seleccionaron 63 artículos para la revisión. Resultados: la evidencia sugiere que el SARS-CoV-2 tiene tropismo no solo limitado a las vías respiratorias. La progresión clínica de la COVID-19 presenta un curso bifásico, con manifestaciones de tipo gripal en la primera fase y episodios postagudos y persistentes en la fase tardía, ocasionados por el daño directo al sistema nervioso central, cardiovascular, endocrino y renal. Conclusiones: la infección por SARS-CoV-2 no debe considerarse solo como una infección aguda y circunscrita a las vías respiratorias.

RESUMO Objetivo: Como a infecção por SARS-CoV-2 pode afetar gravemente os pulmões e podem ocorrer alterações funcionais persistentes após doença grave, nosso objetivo foi determinar os parâmetros de função pulmonar de pacientes com COVID-19, 45 dias após a alta hospitalar e comparar as alterações de acordo com a gravidade da doença. Métodos: Estudo multicêntrico, prospectivo, descritivo e analítico. Os participantes foram alocados em três grupos: grupo internação em enfermaria (IE); grupo internação em UTI sem ventilação mecânica (UTI/VM−); e grupo internação em UTI com VM (UTI/VM+). Volumes pulmonares, DLCO, PImáx, PEmáx e distância percorrida no teste de caminhada de seis minutos (DTC6) foram medidos 45 dias após a alta. Resultados: A amostra foi composta por 242 pacientes (média de idade = 59,4 ± 14,8 anos; 52,1% do sexo masculino), e 232 (96%) apresentavam alteração da função pulmonar. Na coorte total, observou-se distúrbio restritivo em 96%, assim como redução da DLco (em 21,2% dos pacientes), VEF1/CVF (em 39,7%) e PEmáx (em 95,8%), sem diferença entre os grupos para todos. Na comparação entre os grupos, o grupo UTI/VM+ apresentou DLCO reduzida em 50% dos pacientes (p < 0,001) e teve menor média da DTC6 do valor previsto (p = 0,013). Dessaturação de oxigênio no teste de caminhada de seis minutos foi observada em 32,3% da coorte e foi menos frequente no grupo IE. Conclusões: Este é o primeiro estudo sul-americano envolvendo sobreviventes de COVID-19 grave cuja função pulmonar foi avaliada 45 dias após a alta hospitalar. Alterações foram frequentes, principalmente naqueles em VM, o que destaca a importância da avaliação da função pulmonar após COVID-19 grave.

ABSTRACT Objective: Because SARS-CoV-2 infection can severely affect the lungs and persistent functional changes can occur after severe disease, we aimed to determine lung function parameters of COVID-19 patients at 45 days after hospital discharge and compare changes according to the severity of the disease. Methods: This was a prospective descriptive analytical multicenter study. The participants were allocated into three groups: ward admission (WA) group; ICU admission not on mechanical ventilation (ICU/MV−) group; and ICU admission on MV (ICU/MV+) group. Lung volumes, DLco, MIP, MEP, and six-minute walk distance (6MWD) were measured 45 days after discharge. Results: The sample comprised 242 patients (mean age = 59.4 ± 14.8 years; 52.1% of males), and 232 (96%) had altered lung function. In the total cohort, restrictive disorder was observed in 96%, as well as reductions in DLco (in 21.2% of the patients), FEV1/FVC (in 39.7%), and PEmax (in 95.8%), with no differences between the groups. Comparing the groups, the ICU/MV+ group had reduced DLco in 50% of the patients (p < 0.001) and a lower mean 6MWD % of the predicted value (p = 0.013). Oxygen desaturation in the six-minute walk test was observed in 32.3% of the cohort and was less frequent in the IE group. Conclusions: This is the first South American study involving severe COVID-19 survivors whose lung function was assessed 45 days after hospital discharge. Changes were frequent, especially in those on MV, which highlights the importance of lung function evaluation after severe COVID-19.

ABSTRACT Covid-19 disease shows many different manifestations, from common symptoms affecting both the upper and lower respiratory tract, and also sensorial alterations such as anosmia or taste perversion. Heart valve thrombosis and subsequent endocarditis are extremely rare manifestations and they have been scarcely described within the context of Covid-19. We present a case of a patient that was positive for SARS-Cov-2, who developed Staphylococcus hominis bacteremia. Afterwards, he developed persistent fever, and after ruling out some differential diagnoses, a diagnosis of infective endocarditis affecting a native healthy aortic valve was made, using cardiac ultrasonography. This occurrence was considered to be hospital-acquired.

RESUMEN La enfermedad del COVID-19 presenta actualmente una gama de síntomas que van desde la presencia de síntomas clásicos de compromiso de las vías respiratorias altas y bajas hasta alteraciones sensitivas como anosmia o trastornos del gusto. Las trombosis valvulares cardíacas y la endocarditis subsecuente son presentaciones extremadamente raras y poco descritas de la infección por COVID-19. Presentamos el caso de un paciente positivo para virus SARS-Cov2, quien desarrolló una bacteriemia por Staphylococcus hominis y que, ante episodios febriles persistentes y posterior evaluación de diagnósticos diferenciales, se identificó el diagnóstico de endocarditis infecciosa en válvula aórtica nativa sana -mediante ecocardiografía-, la cual fue adquirida en el hospital.

RESUMO Objetivo: Determinar os resultados da ceratoplastia penetrante (PK) para o tratamento da cicatriz da córnea consequente à ceratite por Herpes simplex vírus (HSV), e se o tipo de cicatriz na córnea afeta o resultado cirúrgico. Métodos: Foi realizada análise retrospectiva dos pacientes, submetidos à PK para a cicatriz da córnea relacionados com o HSV entre janeiro de 2008 e julho de 2011. Os pacientes foram divididos em dois grupos. Grupo 1 consistiu de pacientes que tiveram cicatriz corneana herpética quiescente e grupo 2 consistiu de pacientes que desenvolveram descemetocele ou perfuração córnea secundária a defeitos epiteliais persistentes sem inflamação estromal ativa. O seguimento médio foi de 21,30 ± 14,59 meses. Os principais parâmetros avaliados foram recorrência de ceratite herpética, rejeição de enxerto, falência do enxerto, acuidade visual e taxa de sobrevida do enxerto. Resultados: Foram avaliados 42 pacientes do grupo 1 e 13 doentes do grupo 2. Acuidade visual pré-operatória variou de movimentos das mãos (HM) para 0,7 logMAR. No pós-operatório, 34 pacientes (61,8%) atingiram acuidade visual de 0,6 logMAR ou melhor. Doze olhos (28,57%) tiveram recorrência de HSV ceratite no grupo 1, e quatro olhos (30,76%) tiveram recorrência no grupo 2 (p=0,40). A rejeição do enxerto ocorreu em 4 olhos (9,52%) no grupo 1, e em 3 olhos do grupo 2 (23,07%; p=0,58), taxa de sobrevivência do enxerto foi de 91,9% a 1 ano, 76,0% aos 2 anos e 65,1% aos 3 anos no grupo 1, e 89,5% a 1 ano, 76,0% aos 2 anos e 63,6% aos 3 anos no grupo 2 (p=0,91). Conclusões: Embora diferentes taxas de recorrência e de rejeição do enxerto foram encontradas nos dois grupos, a taxa de sobrevida do enxerto em 3 anos foi semelhantes nos dois grupos. De acordo com nossos resultados, em casos sem inflamação, a cicatriz herpética da córnea com descemetocele ou perfuração demonstra as taxas de sobrevivência do enxerto semelhantes às da cicatriz corneana herpética quiescente.

ABSTRACT Purpose: To determine the outcomes of penetrating keratoplasty (PK) for treatment of corneal scarring caused by Herpes simplex virus (HSV) keratitis, and whether the corneal scar type affects treatment outcome. Methods: A retrospective analysis of patients who underwent PK for HSV-related corneal scarring between January 2008 and July 2011 was performed. The patients were categorized into two groups. Group 1 consisted of patients with a quiescent herpetic corneal scar and group 2 consisted of patients who developed a corneal descemetocele or perforation secondary to persistent epithelial defects with no active stromal inflammation. The mean follow-up was 21.30 ± 14.59 months. The main parameters evaluated were recurrence of herpetic keratitis, graft rejection, graft failure, visual acuity, and graft survival rate. Results: There were 42 patients in group 1 and 13 in group 2. Preoperative BCVA varied from hand movements to 0.7 logMAR. Postoperatively, 34 patients (61.8%) achieved visual acuity of 0.6 logMAR or more. Recurrence of HSV keratitis was noted in 12 (28.57%) eyes in group 1 and 4 (30.76%) eyes in group 2 (p=0.40). Graft rejection occurred in 4 eyes (9.52%) in group 1 and in 3 (23.07%) eyes in group 2 (p=0.58). The 1-, 2-, and 3-year graft survival rates were 91.9%, 76.0%, and 65.1% in group 1, and 89.5%, 76.0%, and 63.6% in group 2 (p=0.91), respectively. Conclusions: Although there were different recurrence and graft rejection rates for two groups, the graft survival rates at 3 years were similar. According to our results, without inflammation, corneal herpetic scarring with a descemetocele or perforation achieved similar graft survival rates with quiescent herpetic corneal scars.

ABSTRACT Objective: From December 2013 to March 7, 2015, a total of 22 283 confirmed/probable cases and 863 207 suspected cases of chikungunya were reported in the Caribbean (1). More information regarding symptoms and protection factors is necessary to accurately inform the public and have effective case management. The purpose of this study was to assess age group differences with respect to chikungunya (CHIKV) symptoms and explore protection differences of people who have and have not had CHIKV. Methods: A cross-sectional study sampling 154 participants was conducted from April to May 2015. Participants completed a 37-item questionnaire and were recruited from public places and at a local retirement association in Grenada. Results: Knee joint pain (p < 0.05) and persisting symptoms (rash p < 0.05, muscle pain p < 0.005, ankle joint pain, p < 0.05) were statistically significant for all age groups. No statistical difference was found for taking precautions before and after the outbreak for those who did and did not have CHIKV. However, precaution comparisons for the home (p < 0.001) and work (p < 0.001) environments showed significant non-change before and after the outbreak. Conclusion: This study found that age was a factor for symptom development and persistence, and resistance to behaviour change is an important factor for future epidemic responses and intervention studies.

RESUMEN Objetivo: De diciembre de 2013 al marzo 7 de 2015, un total de 22,283 casos probables/confirmados, y 863,207 casos sospechosos fueron reportados en el Caribe (1). Se requiere más información sobre los síntomas y factores de protección para informar al público con exactitud y lograr un manejo eficaz de los casos. El propósito de este estudio fue evaluar las diferencias de los grupos por edad con respecto a los síntomas de la fiebre chikunguña (CHIKV) y explorar las diferencias de protección entre las personas que han tenido y no han tenido CHIKV. Métodos: Un estudio transversal qué tomó 154 participantes como muestras, se realizó de abril a mayo de 2015. Los participantes completaron un cuestionario de 37 ítems y fueron reclutados de lugares públicos y una asociación local de retiro en Granada. Resultados: El dolor en las articulaciones de la rodilla (p < 0.05) y los síntomas persistentes (erupción p < 0.05, dolor muscular p < 0.005, dolor en las articulaciones tobillo, p < 0.05) fueron esta-dísticamente significativos para todas las edades. No se encontró ninguna diferencia estadística significativa en cuanto a tomar precauciones antes y después del brote, tanto para aquellos que tuvieron como para los que no tuvieron CHIKV. Sin embargo, las comparaciones de precaución con respecto al hogar (p < 0.001) y el entorno de trabajo (p < 0.001) mostraron una significativa ausencia de cambios antes y después del brote. Conclusión: Este estudio encontró que la edad fue un factor para el desarrollo y persistencia de síntomas, y la resistencia al cambio de comportamiento es un factor importante para futuras respuestas epidémicas y estudios de intervención.

Herpes simplex keratitis is one of the leading causes of infectious corneal blindness in the world. It remains latent in the human host after the primary infection and can be reactivated by many factors. When activated, it travels along the trigeminal nerve to the cornea, and causes recurrent infection which leads to corneal scarring. Clinically, preserved AM used as a basement membrane substitute has been applied successfully for the management of persistent epithelial defects with and without ulcerations. Furthermore, multilayered AMT has been described to be useful for the treatment of deep corneal ulcers, descemetoceles, and small corneal perforations. This report describes a case of herpes simplex keratitis treated by amniotic membrane transplantation that resulted in high visual acuity and the absence of keratitis recurrent during two years

La queratitis por herpes simple es una de las causas más importantes de ceguera por infección corneal. El virus permanece latente en el huésped humano después de la infección primaria y puede ser reactivado por muchos factores, como traumatismos, estrés, exposición al sol, procesos que cursen con fiebre asociada, procedimientos quirúrgicos, entre otros. Una vez activado, se traslada a través del nervio trigémino hacia la córnea y causa la infección recurrente, la cual produce cicatrización corneal. Clínicamente, la membrana amniótica usada como sustituto de la membrana basal ha sido aplicada satisfactoriamente para el manejo de defectos epiteliales persistentes con o sin ulceración. Además, se ha descrito que el trasplante de membrana amniótica de múltiples capas es útil para el tratamiento de úlceras corneales profundas, descemetoceles y perforaciones corneales pequeñas. El presente reporte describe un caso de queratitis por herpes simple tratado con membrana amniótica con el que se obtuvo una agudeza visual buena y ausencia de queratitis recurrente durante dos años

Introduction: Polyomavirus BK (BKPyV) and JC (JCPyV) are persistent pathogens able to reactivate in im-munocompromised patients, involving mostly urinary and central nervous system. There are no Chilean studies in HIV positive patients. Objective: To detect BKPyV and JCPyV in blood of Chilean HIV positive adult patients and to correlate these results with clinical-related variables. Materials and Methods: 96 stored blood samples from HIV patients belonging to the north area of Santiago were analyzed. Viral genomes of both viruses were detected by real-time PCR. For statistical analysis, chi-square (Pearson) and Mann-Whitney tests were used and p-values < 0.05 were considered significant. Results: 33% of the samples were positive for BKPyV and a significant correlation was found between the presence of BKPyV genome and the absence of detectable HIV viral load. We demonstrated the need to consider more than one amplification target to detect the BKPyV genome. All the samples were negative for JCPyV genome. Discussion: BKPyV prevalence in Chilean HIV patients is higher than most of international studies. New studies regarding the interaction between both viruses are required. These patients should undergo periodic evaluations by urologist and nephrologist.

Introducción: Polyomavirus BK (BKPyV) y JC (JCPyV) son patógenos persistentes con capacidad de reactivación en inmunocomprometidos, afectando principalmente el sistema urinario y el sistema nervioso central, respectivamente. No existen estudios chilenos en población infectada por VIH. Objetivo: Detectar la presencia de BKPyV y JCPyV en muestras de sangre de pacientes adultos, chilenos, con infección por VIH y correlacionar los resultados con variables clínicas. Materiales y Métodos: Analizamos 96 muestras de extractos leucocitarios de pacientes del área norte de Santiago. El genoma viral se detectó mediante RPC en tiempo real. Para el análisis estadístico se utilizó las pruebas chi-cuadrado de Pearson y Mann-Whitney, considerando significativo un valor de p < 0,05. Resultados: 33% de las muestras resultaron positivas para BKPyV y se encontró una correlación significativa entre la presencia de genoma de BKPyV y la ausencia de carga viral de VIH. Se evidenció la necesidad de considerar más de un blanco de amplificación del genoma de BKPyV. Todas las muestras fueron negativas para JCPyV. Discusión: La prevalencia de BKPyV en pacientes chilenos con infección por VIH es superior a la informada en la mayoría de los reportes internacionales. Se requiere estudios que evalúen la interacción entre ambos virus. Estos pacientes deberían ser sometidos a controles urológicos y nefrológicos periódicos.

Biotechnical of assisted reproduction have become commercially important in recent years. Associated with these techniques the question about his possible role in the transmission of infectious agents via semen and oocytes used as starting material arises. The high prevalence and ability to cause reproductive infections of the bovine viral diarrhea virus (BVDV) has become a potential problem in the in vitro production embryos. This update addresses the previous data on the subject and shows own results on the ways of infection with bovine viral diarrhea (BVDV) associated with in vitro production techniques and their impact on the reproduction bovine. The results demonstrate changes in the ovarian follicular development associated with persistent BVDV infection. These changes were reflected in the low efficiency obtained in the in vitro fertilization as a result of early interaction between the virus and ovarian germ line. Moreover, it was determined that noncytopathic BVDV can cross the zona pellucida of bovine oocytes infected at the beginning of the stage of in vitro maturation. This finding highlights the importance of to understand fully the risks of transmission of virus via semen and oocytes and appropriate quality assurances are used in in vitro production embryos systems (IVP). Likewise, the association of BVDV was demonstrated with gametes when IVF was performed with both types from animals infected persistent, where a decrease was observed in cleavage rates and embryo development. The information obtained in this review article contributes to knowledge about the implications of BVDV infections in IVP systems and its effect on embryonic development, as well as the impact of the transmission of infection in cattle by breeding techniques assisted.

Las biotécnicas de reproducción asistida han adquirido importancia comercial en los últimos años. Asociado a estas técnicas surge el interrogante sobre su rol en la posible transmisión de agentes infecciosos vía semen y ovocitos utilizados como material de partida. La elevada prevalencia y capacidad de causar infecciones reproductivas del virus de la Diarrea Viral Bovina (vDVB) lo ha convertido en un problema potencial en la producción in vitro de embriones. La presente revisión aborda los antecedentes previos sobre la temática y muestra resultados propios sobre las vías de infección del virus de la Diarrea Viral Bovina (vDVB) asociadas a las técnicas de producción in vitro (PIV) y su impacto en la reproducción bovina. Los resultados obtenidos demuestran alteraciones en el desarrollo folicular ovárico asociadas a infecciones persistentes por vDVB. Estas alteraciones fueron reflejadas en la baja eficiencia obtenida en la fecundación in vitro (FIV) como resultado de la interacción temprana entre el virus y la línea germinal del ovario. Por otra parte, se determinó que el vDVB-no citopático (vDVB-ncp) puede atravesar la zona pelúcida de ovocitos bovinos infectados al inicio de la etapa de maduración in vitro (MIV). Este hallazgo pone en evidencia la importancia de controles sanitarios en los sistemas de producción in vitro basados en la comprensión de los riesgos de transmisión del virus a partir de semen y ovocitos. Asimismo, la asociación del vDVB con gametas fue demostrada cuando la FIV se realizó tanto con ovocitos como con semen de animales persistentemente infectados (PI), donde se observó una disminución en las tasas de división y desarrollo de embriones. La información presentada en este artículo de revisión aporta al conocimiento sobre las implicancias de infecciones por vDVB en los sistemas de PIV y su efecto en el desarrollo embrionario, como así también al impacto de la transmisión de la infección en el ganado bovino mediante técnicas de reproducción asistida.

Objective. To describe a political mapping on discrimination and homophobia associated to human immunodeficiency virus (HIV) in the context of public institutions in Mexico. Materials and methods. The political mapping was conducted in six Mexican states. Stakeholders who were involved in HIV actions from public and private sectors were included. Semistructured interviews were applied to explore homophobia and discrimination associated with HIV. Information was systematized using the Policy Maker software, which is a good support for analyzing health policies. Results. Discriminatory and homophobic practices in the public domain occurred, damaging people´s integrity via insults, derision and hate crimes. Most stakeholders expressed a supportive position to prevent discrimination and homophobia and some of them had great influence on policy-making decisions. It was found that state policy frameworks are less specific in addressing these issues. Conclusions. Homophobia and discrimination associated to HIV are still considered problematic in Mexico. Homophobia is a very sensitive issue that requires further attention. Also, an actual execution of governmental authority requires greater enforcement of laws against discrimination and homophobia.

Objetivo. Presentar un mapeo político sobre discriminación y homofobia asociadas con la epidemia del virus de la inmunodeficiencia humana (VIH) en entornos institucionales públicos. Material y métodos. Se realizó un mapeo político con actores clave en seis estados de México, a quienes se les aplicó una entrevista semiestructurada para explorar la discriminación y homofobia asociadas con el VIH. La información se sistematizó mediante matrices de contenido por categorías y el software PolicyMaker. Resultados. Se documentaron prácticas discriminatorias y homofóbicas lesivas a la integridad como la denostación, la burla y los crímenes de odio. La mayoría de actores se manifestaron en contra de la discriminación y la homofobia; algunos de ellos influyeron de manera importante en la formulación de políticas para prevenir la discriminación y la homofobia. Sin embargo, los marcos normativos estatales fueron poco específicos. Conclusiones. Son persistentes las prácticas discriminatorias y la homofobia asociadas con el VIH, por lo cual se requiere mayor atención en los estados y es necesario impulsar políticas para prevenirlas, así como la observancia del cumplimiento de las leyes estatales.

Objective. To describe a political mapping on discrimination and homophobia associated to human immunodeficiency virus (HIV) in the context of public institutions in Mexico. Materials and methods. The political mapping was conducted in six Mexican states. Stakeholders who were involved in HIV actions from public and private sectors were included. Semistructured interviews were applied to explore homophobia and discrimination associated with HIV. Information was systematized using the Policy Maker software, which is a good support for analyzing health policies. Results. Discriminatory and homophobic practices in the public domain occurred, damaging people´s integrity via insults, derision and hate crimes. Most stakeholders expressed a supportive position to prevent discrimination and homophobia and some of them had great influence on policy-making decisions. It was found that state policy frameworks are less specific in addressing these issues. Conclusions. Homophobia and discrimination associated to HIV are still considered problematic in Mexico. Homophobia is a very sensitive issue that requires further attention. Also, an actual execution of governmental authority requires greater enforcement of laws against discrimination and homophobia.

Objetivo. Presentar un mapeo político sobre discriminación y homofobia asociadas con la epidemia del virus de la inmunodeficiencia humana (VIH) en entornos institucionales públicos. Material y métodos. Se realizó un mapeo político con actores clave en seis estados de México, a quienes se les aplicó una entrevista semiestructurada para explorar la discriminación y homofobia asociadas con el VIH. La información se sistematizó mediante matrices de contenido por categorías y el software PolicyMaker. Resultados. Se documentaron prácticas discriminatorias y homofóbicas lesivas a la integridad como la denostación, la burla y los crímenes de odio. La mayoría de actores se manifestaron en contra de la discriminación y la homofobia; algunos de ellos influyeron de manera importante en la formulación de políticas para prevenir la discriminación y la homofobia. Sin embargo, los marcos normativos estatales fueron poco específicos. Conclusiones. Son persistentes las prácticas discriminatorias y la homofobia asociadas con el VIH, por lo cual se requiere mayor atención en los estados y es necesario impulsar políticas para prevenirlas, así como la observancia del cumplimiento de las leyes estatales.