Whole Genome Sequence-Based Analysis of Bovine Gammaherpesvirus 4 Isolated from Bovine Abortions.

Bovine gammaherpesvirus 4 (BoGHV4) is a member of the Gammaherspivirinae subfamily, Rhadinovirus genus. Its natural host is the bovine, and it is prevalent among the global cattle population. Although the complete genome of BoGHV4 has been successfully sequenced, the functions of most of its genes remain unknown. Currently, only six strains of BoGHV4, all belonging to Genotype 1, have been sequenced. This is the first report of the nearly complete genome of Argentinean BoGHV4 strains isolated from clinical cases of abortion, representing the first BoGHV4 Genotype 2 and 3 genomes described in the literature. Both Argentinean isolates presented the highest nt p-distance values, indicating a greater level of divergence. Overall, the considerable diversity observed in the complete genomes and open reading frames underscores the distinctiveness of both Argentinean isolates compared to the existing BoGHV4 genomes. These findings support previous studies that categorized the Argentinean BoGHV4 strains 07-435 and 10-154 as Genotypes 3 and 2, respectively. The inclusion of these sequences represents a significant expansion to the currently limited pool of BoGHV4 genomes while providing an important basis to increase the knowledge of local isolates.

Patient safety topics, especially the second victim phenomenon, are neglected in undergraduate medical and nursing curricula in Europe: an online observational study.

This study aims to assess the inclusion of second victims and other patient safety issues in the curricula of undergraduate medical and nursing degrees in the countries participating in the European Researchers' Network Working on Second Victims (The ERSNT Consortium, COST Action 19,113). A review of medical and nursing school curricula in 206 universities was carried out, using their websites to search for subjects addressing "patient safety", "quality of care", "risk management", "safe practices", "interprofessional communication", "adverse events", and "second victims". There was substantial variability in the extent of training for patient safety. Forty-four out of 88 nursing schools and 74 of 118 medical schools did not include any of the patient safety topics studied. The most frequent in both nursing and medicine was "interprofessional communication", followed by "quality of care" and basic aspects on "patient safety". The second victim phenomenon was present in only one curriculum of the total sample. Our study showed that patient safety, especially the second victim phenomenon, is still neglected in medical and nursing curricula in European universities, although positive initiatives were also found. Given the frequency with which adverse events occur in health centres and the need to prepare students to deal with them adequately, additional efforts are needed to introduce patient safety elements into medical and nursing education.

Study of the dynamics of in vitro infection with bovine gammaherpesvirus type 4 and apoptosis markers in susceptible cells.

Bovine gammaherpesvirus type 4 (BoHV-4) shows tropism for the endometrium, in which it causes the death of epithelial and stroma cells. Despite having anti-apoptotic genes in its genome, experiments based on immortalized cell lines have shown that BoHV-4 induces cell death by apoptosis. In the present study, we evaluated BoHV-4 replication, pro-apoptotic (Bax) and anti-apoptotic (Bcl-2) mitochondrial genes expression and chromatin condensation in bovine endometrium primary culture cells (BEC) and in the Madin Darby bovine kidney (MDBK) cell line. Results showed that BoHV-4 has a preference for replication in BEC cells over the MDBK cell line, demonstrated by the high viral titer that is consistent with the tropism of the virus. In BEC cells, chromatin condensation was consistent with the values of viral kinetics at the late stage of infection, accompanied with a balance in the mRNA levels of apoptotic mitochondrial proteins. As a consequence, in those cells viral transmission would be enhanced by inhibiting apoptosis in the early stage of virus proliferation, allowing the complete production of viral progeny, and then, the induction of apoptosis in late stages would allow neighboring cells infection. In MDBK cells replication kinetics was coincident with the up-regulation of Bcl-2, which suggests that the productive infection in MDBK is associated with a lytic phase of the virus or another cell death pathway (probably autophagy mechanism) at the late stage of infection. The results agree with the study of nuclear morphology, where a constant chromatin condensation was observed over time. It is clear that the documented BoHV-4 apoptotic responses observed in the cell lines studied above are not valid in cells from primary cultures. The data presented in this study suggest that BoHV-4 could induce apoptosis in BEC cells without a leading role of the mitochondria pathway. Further studies will be necessary to characterize in detail the programmed cell death pathways involved in BoHV-4 infection in the primary cell cultures evaluated.

Chitosan-Based Formulations Intended as Protective Spray for Mask Surfaces in Prevention of Coronavirus Dissemination.

The extraordinary occurrence of COVID-19 by the fast expansion of viral infections has propelled particular interest in developing novel antiviral and virucidal agents to guarantee personal security. The main objective of this work is to propose novel formulations able to optimize the use of personal protection elements. In recent years, chitosan (CH) has attracted attention for being an interesting multifunctional, biodegradable, non-antigenic, non-toxic, and biocompatible natural polymer with antimicrobial properties. In this work, formulations based on a CH matrix containing silver, and Copper based nanoparticles have been developed. The novelty of this proposal is that almost liquid formulations have been reached, possessing verified properties to inhibit evolved virus such as herpes simplex type 1 (HSV-1) and bovine betacoronavirus (BCoV), the latter belonging to the same family of the well-known the well-known SARS-CoV-2. Besides antibacterial bioactivity; as well as the ability of these formulations to be easily sprayed on various surfaces, including conventional face masks, have been verified and discussed. The results presented in this contribution provide strong evidence on CH films as an ideal biosafe surface-protective for several daily used materials including the conventional face masks.

Genetic characterization of bovine herpesvirus 4 (BoHV-4) isolates from Argentine cattle suggests a complex evolutionary scenario.

Bovine herpevsirus 4 (BoHV-4) is a gammaherpesvirus that has been associated with different clinical conditions in cattle. In Argentina, BoHV-4 was detected in diverse bovine samples. The aim of this study was to analyze the genetic relationship of 48 field BoHV-4 strains isolated from cattle in Argentina. According to thymidine kinase (tk) gene sequences, BoHV-4 isolates belong to genotypes 1, 2 and 3. Phylogenetic analyses confirmed the presence of the three previously described viral genotypes. However, some of the studied isolates presented conflicting phylogenetic signals between the studied markers. This suggests a complex evolutionary background, that is a history of recombination, incomplete lineage sorting (deep coalescence) or a combination of these, which requires further study. These potential events make difficult the diagnosis of BoHV-4 from clinical samples of cattle and may pose a significant problem for the control of the virus in the herds.

Analysis of the anti-apoptotic v-Bcl2 and v-Flip genes and effect on in vitro programmed cell death of Argentinean isolates of bovine gammaherpesvirus 4 (BoHV-4).

Some viruses encode inhibitory factors of apoptosis during infection to prolong cell viability and then to achieve a higher production of viral progeny or facilitate persistent infections. There is evidence that some gammaherpesviruses, including BoHV-4, carry genes that can both inhibit or induce apoptosis. BoHV-4 possesses two genes (ORF16 and ORF71) that code for proteins with anti-apoptotic functions, such as v-Bcl2 and v-Flip, respectively. Thus, it is relevant to study BoHV-4 in relation to the modulation of apoptosis in infected cells as a strategy for persistence in the host. The objective of this work was to analyze whether variations in v-Flip and v- Bcl2 of six phylogenetically divergent Argentinean isolates of BoHV-4 can influence the capacity of these strains to induce apoptosis in cell cultures. In this study, variations were mainly detected in the v-Flip gene and protein of the BoHV-4 strains belonging to genotype 3. Thus, it is possible to infer that sequence variations could be associated with some BoHV-4 genotype. Induction of apoptosis was not a significant event for any of the genetically distinct local isolates of BoHV-4 and there was not an evident relationship between the variability of both genes with the apoptotic effect of the phylogenetically distinct strains.

Analysis of the transcripts encoding for antigenic proteins of bovine gammaherpesvirus 4.

The major glycoproteins of bovine gammaherpesvirus 4 (BoHV-4) are gB, gH, gM, gL, and gp180 with gB, gH, and gp180 being the most glycosylated. These glycoproteins participate in cell binding while some act as neutralization targets. Glycosylation of these envelope proteins may be involved in virion protection against neutralization by antibodies. In infected cattle, BoHV-4 induces an immune response characterized by low neutralizing antibody levels or an absence of such antibodies. Therefore, virus seroneutralization in vitro cannot always be easily demonstrated. The aim of this study was to evaluate the neutralizing capacity of 2 Argentine BoHV-4 strains and to associate those findings with the gene expression profiles of the major envelope glycoproteins. Expression of genes coding for the envelope glycoproteins occurred earlier in cells infected with isolate 10/154 than in cells infected with strain 07/435, demonstrating a distinct difference between the strains. Differences in serological response can be attributed to differences in the expression of antigenic proteins or to post-translational modifications that mask neutralizing epitopes. Strain 07/435 induced significantly high titers of neutralizing antibodies in several animal species in addition to bovines. The most relevant serological differences were observed in adult animals. This is the first comprehensive analysis of the expression kinetics of genes coding for BoHV-4 glycoproteins in 2 Argentine strains (genotypes 1 and 2). The results further elucidate the BoHV-4 life cycle and may also help determine the genetic variability of the strains circulating in Argentina.

Comparative analysis of replicative properties of phylogenetically divergent, Argentinean BoHV-4 strains in cell lines from different origins.

Bovine gammaherpesvirus 4 (BoHV4) is a member of the family Herpesviridae. In Argentina, BoHV4 was isolated and characterized in 2007 from samples of aborted cows. Argentinean isolates are highly divergent and are classified as: Genotype 1(Movar-like), Genotype 2 (DN599-like) and Genotype 3 (a novel group). The aim of this study was to comparatively evaluate the biological characteristics of six Argentinean BoHV4 field isolates in cell lines from different origins. All strains induced productive infection in the cell lines used, with different degrees of permissiveness. A direct relationship among the times of appearance of cytopathic effect, the growth kinetics, the size of the lysis plaques and the virulent-like behaviour in vitro could not be established. However, although slight, there are differences in the biological behaviour of the BoHV4 fields isolates analyzed. This variability is independent of their genetic classification but would be conditioned by the nature of the infected cells.

Bovine herpesvirus type 5 replication and induction of apoptosis in vitro and in the trigeminal ganglion of experimentally-infected cattle.

Bovine herpesvirus (BoHV) types 1 and 5 are neuroinvasive. Cases of BoHV-1-induced encephalitis are not as frequent as those caused by BoHV-5. In this study, the capability of BoHV-5 to induce apoptosis in cell cultures and in the trigeminal ganglion during acute infection of experimentally-infected cattle was analyzed. Apoptotic changes in cell cultures agree with the ability of the viral strains to replicate in each cell line. Marked differences were observed between the in vitro induction of apoptosis by BoHV-1Cooper and BoHV-5 97/613 strains. Apoptotic neurons were clearly evident in the trigeminal ganglion of BoHV-1-infected calves. For BoHV-5 a fewer number of positive neurons was observed. There is an association between the magnitude of bovine herpesviruses replication and the induction of apoptosis in trigeminal ganglion. These findings suggest that the induction of apoptosis and the innate immune response orchestrate the final outcome of alpha herpesviruses infection of the bovine nervous system.

Involvement of toll-like receptors 3 and 7/8 in the neuropathogenesis of bovine herpesvirus types 1 and 5.

Bovine herpesvirus types 1 (BoHV-1) and 5 (BoHV-5) are closely related alpha-herpesviruses. BoHV-5 is the causal agent of non-suppurative meningoencephalitis in calves. BoHV-1 causes respiratory disease, abortions, genital disorders and, occasionally, encephalitis in cattle. Both viruses are neurotropic and they share similar biological properties. Nevertheless, they differ in their ability to cause neurological disease. Toll-like receptors (TLRs) are involved in the innate immune response to pathogens. In this study, the variations in the expression levels of TLRs were evaluated in different regions of the bovine central nervous system during the acute infection and reactivation of BoHV-1 and BoHV-5- infected cattle. With the exception of TLR9, significant up-regulation of all TLRs was detected following primary infection of neural tissues by both bovine alpha-herpesviruses. Furthermore, the stages of acute infection and reactivation were characterized by a distinguishable TLR expression pattern. Important differences in TLR expression upon infection of the central nervous system by BoHV-1 or BoHV-5 were not detected. The striking differences in TLR mRNA levels during acute infection and reactivation provide evidence that the innate immune response may be involved in the clinical outcomes observed at each stage. Further research is required to analyze the mechanisms that initiate TLR activation and the signaling cascade mediated by each TLR to elucidate the precise role these receptors play in bovine herpesvirus encephalitis.

Herpesvirus bovino 4 (BoHV-4): aspectos generales de su biología y situación en la República Argentina / Bovine herpesvirus 4 (BoHV-4): general aspects of the biology and status in Argentina

El herpesvirus bovino 4 [Bovine herpesvirus 4 (BoHV-4)] ha sido aislado de bovinos con infecciones respiratorias, vulvovaginitis, mastitis, abortos, endometritis y de animales aparentemente sanos en diferentes partes del mundo. Si bien no se ha reconocido como agente causal de una entidad patológica en particular, se asocia principalmente con infecciones del tracto reproductivo de los bovinos. Este virus puede infectar un amplio rango de especies tanto in vivo como in vitro. Los primeros aislamientos dieron origen a dos grupos de cepas prototipo: el grupo americano tipo DN599 y el grupo europeo tipo Movar. En Argentina, el BoHV-4 fue aislado y caracterizado en el año 2007; este aislamiento se obtuvo de muestras de mucus cérvico-vaginal de vacas que abortaron. Hasta el momento se han registrado más de 40 aislamientos, provenientes principalmente de hembras bovinas que han abortado

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle with respiratory infections, vulvovaginitis, mastitis, abortions, endometritis and from apparently healthy animals throughout the world. Although it has not yet been established as causal agent of a specific disease entity, it is primarily associated with reproductive disorders of cattle. This virus can infect a wide range of species, either in vivo or in vitro. Two groups of prototype strains were originated from the first isolates: the DN599-type strains (American group) and the Movar-type strains (European group). In Argentina, BoHV-4 was isolated and characterized in 2007 from vaginal discharge samples taken from cows that had aborted. So far, more than 40 isolates, mainly associated with aborting bovine females have been registered in our country

[Bovine herpesvirus 4 (BoHV-4): general aspects of the biology and status in Argentina]. / Herpesvirus bovino 4 (BoHV-4): aspectos generales de su biología y situación en la República Argentina.

Bovine herpesvirus 4 (BoHV-4) has been isolated from cattle with respiratory infections, vulvovaginitis, mastitis, abortions, endometritis and from apparently healthy animals throughout the world. Although it has not yet been established as causal agent of a specific disease entity, it is primarily associated with reproductive disorders of cattle. This virus can infect a wide range of species, either in vivo or in vitro. Two groups of prototype strains were originated from the first isolates: the DN599-type strains (American group) and the Movar-type strains (European group). In Argentina, BoHV-4 was isolated and characterized in 2007 from vaginal discharge samples taken from cows that had aborted. So far, more than 40 isolates, mainly associated with aborting bovine females have been registered in our country.

Consenso español de salud física del paciente con depresión / Spanish consensus on the physical health of patients with depressive disorders

La comorbilidad entre depresión y enfermedades físicas es muy frecuente y tiene un impacto importante en el deterioro de la salud así como en la atención y tratamiento recibidos. Impulsado por las Sociedades Españolas de Psiquiatría y Psiquiatría Biológica, y la Sociedad Española de Médicos de Atención Primaria (SEMERGEN) se realizó un consenso sobre la salud física en el paciente con depresión, que de forma resumida se expone en el presente trabajo. La revisión de la literatura puso de manifiesto la elevada frecuencia en pacientes con depresión de trastornos cardiovasculares y trastornos endocrinometabólicos como la obesidad y la diabetes, de tal modo que en pacientes con depresión serían aplicables las recomendaciones de prevención primaria y secundaria establecidas para los individuos con riesgo cardiovascular o metabólico. También es frecuente la comorbilidad entre depresión y dolor crónico, que obliga a la detección activa de ambos trastornos y a una intervención terapéutica integral. La presencia de esta comorbilidad puede condicionar, aunque no impedir el tratamiento farmacológico en cuya elección se deberá tener en cuenta los efectos secundarios y las interacciones de los distintos antidepresivos. Por su parte, la psicoterapia puede contribuir a la recuperación integral del enfermo. Globalmente, en el proceso de atención al paciente con depresión y enfermedades físicas comórbidas es fundamental la coordinación del médico de atención primaria, el psiquiatra y otros especialistas o profesionales implicados (AU)

Comorbidity between depression and physical illnesses is very common and has a significant impact on the health and management of the patient. With the support of the Sociedades Españolas de Psiquiatría y Psiquiatría Biológica, and Sociedad Española de Médicos de Atención Primaria (SEMERGEN) a consensus was prepared on physical health in patients with depression and is summarized in the present work. The literature review highlighted the high frequency of cardiovascular and endocrine-metabolic disorders in patients with depression such as diabetes and obesity, thus making the primary and secondary prevention recommendations for patients with cardiovascular or metabolic risk applicable to patients with depression. Comorbidity between depression and chronic pain is also frequent, and requires an integrated therapeutic approach. The presence of physical illness in patients with depression may condition, but not preclude, the pharmacological treatment; drug selection should take into account potential side-effect and drug-drug interactions. On the other hand, psychotherapy may contribute to the patient's recovery. Overall, coordination between the primary care physician, the psychiatrist and other health professionals involved is essential for the management of patients with depression and concomitant physical illness (AU)

Spanish consensus on the physical health of patients with depressive disorders.

Comorbidity between depression and physical illnesses is very common and has a significant impact on the health and management of the patient. With the support of the Sociedades Españolas de Psiquiatría y Psiquiatría Biológica, and Sociedad Española de Médicos de Atención Primaria (SEMERGEN) a consensus was prepared on physical health in patients with depression and is summarized in the present work. The literature review highlighted the high frequency of cardiovascular and endocrine-metabolic disorders in patients with depression such as diabetes and obesity, thus making the primary and secondary prevention recommendations for patients with cardiovascular or metabolic risk applicable to patients with depression. Comorbidity between depression and chronic pain is also frequent, and requires an integrated therapeutic approach. The presence of physical illness in patients with depression may condition, but not preclude, the pharmacological treatment; drug selection should take into account potential side-effect and drug-drug interactions. On the other hand, psychotherapy may contribute to the patient's recovery. Overall, coordination between the primary care physician, the psychiatrist and other health professionals involved is essential for the management of patients with depression and concomitant physical illness.

Quantitative analysis of campylobacter fetus venerealis adhesion to bovine reproductive tract cell cultures / Análise quantitativa da adesão de campylobacter fetus venerealis em culturas de células do aparelho reprodutor bovino

Campylobacter fetus is the etiological agent of bovine genital campylobacteriosis, a sexually transmitted disease which is associated with reproductive losses in bovines. Campylobacter colonizes the vagina and the uterus and then infects the epithelial cells of the endometrium. The objective of this work was to develop an ex vivo model to quantify the adhesion of Campylobacter to its natural specific target cells; this is a key step for the establishment of infection and studies regarding the adherence and cytotoxicity on the natural host cells are not available. The assays were carried out by seeding Campylobacter fetus venerealis on bovine vaginal and uterine epithelial cell cultures. HeLa cells were used as control. Bacterial adhesion was corroborated by optical microscopy and determination of the percentage of adherent bacteria was performed on immunochemically-stained slides. Results are presented as percentage of cells with adherent Campylobacter and as number of bacteria per cell. In comparison to the control HeLa cells, the statistical analysis revealed that primary cultures show a higher percentage of infected cells and a lower variation of the evaluated parameters. This primary culture model might be useful for studies on cytopathogenicity and adhesion of different field strains of Campylobacter fetus.

Campylobacter fetus é o agente etiológico da campilobacteriose genital bovina, uma doença sexualmente transmissível que está associada com perdas reprodutivas em bovinos. Campylobacter coloniza a vagina e o útero e então infecta as células epiteliais do endométrio. O objetivo deste trabalho foi desenvolver um modelo ex vivo para quantificar a adesão de Campylobacter às células-alvo naturais específicas; este é um passo fundamental para o estabelecimento da infecção e estudos acerca da adesão e citotoxicidade sobre as células do hospedeiro natural não estão disponíveis. Os ensaios foram realizados a través da semeadura de Campylobacter fetus venerealis em culturas celulares epiteliais vaginais e uterinas.Células HeLa foram utilizadas como controle.A aderência bacteriana foi confirmada por microscopia óptica e a determinação da porcentagem de bactérias aderidas foi realizada em lâminas tingidas imunoquimicamente. Os resultados são apresentados como porcentagem de células com Campylobacter aderente e como o número de bactérias por células. Em comparação com as células HeLa controle, a análise estatística revelou que as culturas primárias mostram uma maior porcentagem de células infectadas e uma menor variação dos parâmetros avaliados. Este modelo de cultura primária pode ser útil para estudos sobre citopatogenicidade e adesão de diferentes cepas de campo de Campylobacter fetus.