Parechovirus infection in human brain organoids: host innate inflammatory response and not neuro-infectivity correlates to neurologic disease.

Picornaviruses are a leading cause of central nervous system (CNS) infections. While genotypes such as parechovirus A3 (PeV-A3) and echovirus 11 (E11) can elicit severe neurological disease, the highly prevalent PeV-A1 is not associated with CNS disease. Here, we expand our current understanding of these differences in PeV-A CNS disease using human brain organoids and clinical isolates of the two PeV-A genotypes. Our data indicate that PeV-A1 and A3 specific differences in neurological disease are not due to infectivity of CNS cells as both viruses productively infect brain organoids with a similar cell tropism. Proteomic analysis shows that PeV-A infection significantly alters the host cell metabolism. The inflammatory response following PeV-A3 (and E11 infection) is significantly more potent than that upon PeV-A1 infection. Collectively, our findings align with clinical observations and suggest a role for neuroinflammation, rather than viral replication, in PeV-A3 (and E11) infection.

COVID-19, social determinants of transmission in the home. A population-based study.

BACKGROUND: Studying transmission within the home is essential to understand the transmission dynamics of numerous infectious diseases. For Coronavirus Disease-2019 (COVID-19), transmission within the home constitutes the majority exposure context. The risk of infection in this setting can be quantified by the household/intra-family secondary attack rate (SAR). In the literature, there are discrepancies in these values and little information about its social determinants. The aim of this study was to investigate transmission in the home by analyzing the influence of occupational social class, country of origin and gender/sex. METHODS: This was a retrospective cohort study of a population registry of cohabiting contacts with COVID-19 cases diagnosed from 15 June to 23 December 2020, in the Murcia Region. The household SAR was analyzed considering the characteristics of the primary case (sex, age, symptoms, occupational social class, country of origin and number of people in the household) and contact (age and sex) using a multilevel binary logistic regression model. RESULTS: Among the 37 727 contacts included, the intra-family SAR was 39.1%. The contacts of confirmed primary cases in the migrant population (Africa and Latin America) had higher attack rates, even after adjusting for the other variables. Older age and female sex were independent risk factors for contracting Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) within the home. CONCLUSION: There was greater intra-domiciliary transmission among immigrants, likely related to the conditions of the home and situation of social vulnerability. Women were more likely to be infected by transmission from a cohabiting infected individual.

Assessment of the broad-spectrum host targeting antiviral efficacy of halofuginone hydrobromide in human airway, intestinal and brain organotypic models.

Halofuginone hydrobromide has shown potent antiviral efficacy against a variety of viruses such as SARS-CoV-2, dengue, or chikungunya virus, and has, therefore, been hypothesized to have broad-spectrum antiviral activity. In this paper, we tested this broad-spectrum antiviral activity of Halofuginone hydrobomide against viruses from different families (Picornaviridae, Herpesviridae, Orthomyxoviridae, Coronaviridae, and Flaviviridae). To this end, we used relevant human models of the airway and intestinal epithelium and regionalized neural organoids. Halofuginone hydrobomide showed antiviral activity against SARS-CoV-2 in the airway epithelium with no toxicity at equivalent concentrations used in human clinical trials but not against any of the other tested viruses.

Pavlovian conditioning-induced hallucinations reduce MMN amplitudes for duration but not frequency deviants.

The mismatch negativity (MMN) is an evoked potential that indexes auditory regularity violations. Since the 90's, a reduced amplitude of this brain activity in patients with schizophrenia has been consistently reported. Recently, this alteration has been related to the presence of auditory hallucinations (AHs) rather than the schizophrenia diagnostic per se. However, making this attribution is rather problematic due to the high heterogeneity of symptoms in schizophrenia. In an attempt to isolate the AHs influence on the MMN amplitude from other cofounding variables, we artificially induced AHs in a non-clinical population by Pavlovian conditioning. Before and after conditioning, volunteers (N = 31) participated in an oddball paradigm that elicited an MMN. Two different types of deviants were presented: a frequency and a duration deviant, as the MMN alteration seems to be especially present in schizophrenia with the latter type of deviant. Hence, this pre-post design allowed us to compare whether experiencing conditioning-induced AHs exert any influence on MMN amplitudes. Our results show that duration-deviant related MMN reductions significantly correlate with the number of AHs experienced. Moreover, we found a significant correlation between AHs proneness (measured with the Launay-Slade Hallucination Extended Scale) and the number of AHs experienced during the paradigm. In sum, our study shows that AHs can be conditioned and exert similar effects on MMN modulation in healthy participants as has been reported for patients with schizophrenia. Thus, conditioning paradigms offer the possibility to study the association between hallucinations and MMN reductions without the confounding variables present in schizophrenia patients.

Amino acid variation at VP1-145 of enterovirus A71 determines the viral infectivity and receptor usage in a primary human intestinal model.

Enterovirus A71 (EV-A71) can elicit a wide variety of human diseases such as hand, foot, and mouth disease and severe or fatal neurological complications. It is not clearly understood what determines the virulence and fitness of EV-A71. It has been observed that amino acid changes in the receptor binding protein, VP1, resulting in viral binding to heparan sulfate proteoglycans (HSPGs) may be important for the ability of EV-A71 to infect neuronal tissue. In this study, we identified that the presence of glutamine, as opposed to glutamic acid, at VP1-145 is key for viral infection in a 2D human fetal intestinal model, consistent with previous findings in an airway organoid model. Moreover, pre-treatment of EV-A71 particles with low molecular weight heparin to block HSPG-binding significantly reduced the infectivity of two clinical EV-A71 isolates and viral mutants carrying glutamine at VP1-145. Our data indicates that mutations in VP1 leading to HSPG-binding enhances viral replication in the human gut. These mutations resulting in increased production of viral particles at the primary replication site could lead to a higher risk of subsequent neuroinfection. Importance: With the near eradication of polio worldwide, polio-like illness (as is increasingly caused by EV-A71 infections) is of emerging concern. EV-A71 is indeed the most neurotropic enterovirus that poses a major threat globally to public health and specifically in infants and young children. Our findings will contribute to the understanding of the virulence and the pathogenicity of this virus. Further, our data also supports the identification of potential therapeutic targets against severe EV-A71 infection especially among infants and young children. Furthermore, our work highlights the key role of HSPG-binding mutations in the disease outcome of EV-A71. Additionally, EV-A71 is not able to infect the gut (the primary replication site in humans) in traditionally used animal models. Thus, our research highlights the need for human-based models to study human viral infections.Graphical Abstract.

Analysis of the Chagas disease situation in Japan: a cross sectional study and cost-effectiveness analysis of a Chagas disease screening program.

Background: Japan is estimated to host 3000 cases of Chagas disease (CD). However, there are no epidemiological data and policies for prevention and care. We aimed to analyze the current situation of CD in Japan and identify possible barriers to seeking care. Methods: This cross-sectional study included Latin American (LA) migrants living in Japan from March 2019 to October 2020. We obtained blood samples to identify participants infected with Trypanosoma cruzi, and data about sociodemographic information, CD risk factors, and barriers to access to the Japanese national health care system (JNHS). We used the observed prevalence to calculate the cost-effectiveness analysis of the screening of CD in the JNHS. Findings: The study included 428 participants, most of them were from Brazil, Bolivia and Peru. The observed prevalence was 1.6% (expected prevalence= 0.75%) and 5.3% among Bolivians. Factors associated with seropositivity were being born in Bolivia, having previously taken a CD test, witnessing the triatome bug at home, and having a relative with CD. The screening model was more cost-effective than the non-screening model from a health care perspective (ICER=200,320 JPY). Factors associated with access to JNHS were being female, length of stay in Japan, Japanese communication skills, source of information, and satisfaction about the JNHS. Interpretation: Screening of asymptomatic adults at risk of CD may be a cost-effective strategy in Japan. However, its implementation should consider the barriers that affect LA migrants in access to the JNHS. Funding: Nagasaki University and Infectious Diseases Japanese Association.

Body Composition and Nutritional Status of the Spanish National Breaking Team Aspiring to the Paris 2024 Olympic Games.

Breaking is a sports dance modality that will debut for the first time at the Paris 2024 Olympic Games. This dance form combines street dance steps with acrobatics and athletic elements. It complies with gender equality, maintains aesthetic appeal, and is practised indoors. The objective of this study is to assess the characteristics of body composition and nutritional status of the athletes that make up the Breaking national team. This national team was recruited, and they underwent an analysis of body composition using bioimpedance measurement and a nutritional interview status with the completion of a survey on the frequency of the consumption of sports supplements and ergogenic aids. In addition, they completed a consumption questionnaire for a series of food groups with specified protein, lipid, and carbohydrate content. After that, parameters were analyzed in relation to their nutritional status during a complete medical examination at the Endocrinology and Nutrition Service of the Sports Medicine Center of CSD. A descriptive analysis of the results obtained was carried out to find the mean values of the variables analyzed. The analytical parameters described an adequate nutritional status, except for the mean capillary determination of 25-OH-vitamin D3, which was 24.2 ng/dL (SD: 10.3). Bone mineral density values were higher than those of the general population. This is the first time that a study of these characteristics has been carried out on Breakers, so it is highly relevant to increase knowledge in this area in order to conduct nutritional interventions aimed at improving the sports performance of these athletes.

Impact of Embryo Cryopreservation on Large for Gestational Age Babies Born by Embryo Transfer: Cohort Retrospective Study.

The transfer of frozen-thawed embryos has been associated with an increased risk of large for gestational age at birth. Our objective is to assess its impact on the risk of large for gestational age (LGA) in order to improve the bias control in relation with the available studies. Retrospective cohort study on cumulative sample of 801 single live births of 32 weeks or longer of gestation, resulting from pregnancies obtained by cryopreserved or fresh embryo transfer which are not affected by disorders that could impair fetal growth and carried out at Hospital General Universitario Gregorio Marañón, in Madrid, during the period 2005-2017. The relative risk (RR) of LGA has been estimated with its confidence interval (CI) at 95%. Multivariate analysis using logistic regression was applied to adjust the crude effect. LGA was more frequent in babies born after cryotransfer in comparison with the reference group (20.9% vs. 6.0%; p < 0.001), as well as macrosomia (8.2% vs. 0.9%; p < 0.001). Frequencies of weight > 4500 g were similar (0.7% vs. 0.1%; p > 0.05). Nulliparity was associated to a higher risk of LGA (RR: 3.8; CI95%: 2.0-7.0; p < 0.005), as well as cleaving embryo transfer (RR: 2; CI95%: 1.07-3.8; p < 0.05). According to the multivariate analysis, the exposure variable was the only one independently associated with LGA (OR: 3.5; CI95%: 2.0-6.1; p < 0.001). Frozen-thawed embryo transfer significantly increases the risk of LGA, regardless of the influence of factors relating to the patient's condition, the embryos transferred, or the fetal sex.

[COVID-19 exposure setting, social and gender determinants in a mediterranean region.] / Características sociales y de género en el ámbito de contagio de COVID-19 en una región mediterránea.

OBJECTIVE: Knowledge of social and gender determinants, which influence the places where people are exposed to COVID-19, may be relevant in the development of preventive and control strategies. The aim of this paper was to determine the context in which COVID-19 cases were infected (household, work/labor, health, social-health, and social-leisure settings) according to country of origin, occupational social class and gender, which is essential in order to designing public health strategies. METHODS: A cross-sectional study of an epidemiological registry of 56,628 COVID-19 incident cases was made, whose exposure/contagion setting was studied according to the previous variables from June 15 to December 23, 2020, in the Region of Murcia (Spain). An exact Fisher test was used to study the distribution of COVID-19 cases based on the above variables. RESULTS: The cumulative incidence was higher in people from Africa (5,133.5 cases/100,000 inhabitants) and Latin America (11,351.1) than in non-immigrants (3,145.7). It was also higher in women (3,885.6) than in men (3,572.6). It is noteworthy, that 53.3% of the cases with employment were workers in industry or construction, artisans, agricultural workers, or elementary occupations. In contrast, during the second semester of 2020, 41.3% of the employed population in the Region of Murcia performed such jobs. The household was the main exposure setting (56.5% of cases with a known setting), followed by social-leisure (20.7%) and work/labor (18.2%). The labor settings were more important in immigrants from Africa (28.4%) and Latin America (35.7%) than in non-immigrants (12%), inversely to social-leisure settings. Labor context was more important in women (19.6%) than in men (16.5%) and in manual workers (44.1%) than in non-manual workers (26.6%). CONCLUSIONS: The context in which COVID-19 cases were infected is different according to social inequalities related to country of origin, gender and occupational social class.

OBJETIVO: El conocimiento de los determinantes sociales y de género que influyen en el ámbito de exposición al SARS-CoV-2 puede ser relevante en el planteamiento de estrategias preventivas y de control de la transmisión. No se han encontrado estudios previos que evalúen cómo influyen la clase social ocupacional y el país de origen en el ámbito de exposición del SARS-CoV-2. El objetivo de este trabajo fue conocer el contexto en que se contagiaron los casos de COVID-19 (ámbito: hogar, trabajo, sanitario, sociosanitario y social-ocio) según país de origen, clase social ocupacional y género, lo cual es esencial para orientar estrategias de Salud Pública. METODOS: Se realizó un estudio descriptivo de un registro epidemiológico de 56.628 casos incidentes de COVID-19 en los que se estudió el ámbito de exposición/contagio en función de las variables anteriormente indicadas entre el 15 de junio y el 23 de diciembre de 2020 en la Región de Murcia. Se utilizó una prueba exacta de Fisher para el estudio de la distribución de los casos de COVID-19 en función de las variables anteriores. RESULTADOS: La incidencia acumulada fue mayor en personas procedentes de África (5.133,5 casos por cada 100.000 habitantes) y Latinoamérica (11.351,1) que en no inmigrantes (3.145,7) y superior en mujeres (3.885,6) que en hombres (3.572,6). Es destacable que el 53,3% de los casos COVID-19 con empleo registrado eran operarios en industria o construcción, artesanos, trabajadores agrarios o con ocupaciones elementales (15 de junio y el 23 de diciembre de 2020). En contraste, el 41,3% de la población ocupada en la Región de Murcia realizaba dichos empleos (promedio 3º y 4º trimestre de 2020). El hogar fue el principal ámbito de contagio (56,5% de los casos con ámbito conocido), seguido del social-ocio (20,7%) y el laboral (18,2%). Este último tuvo mayor peso en personas procedentes de África (28,4%) y Latinoamérica (35,7%) que en no inmigrantes (12%), a la inversa que el social-ocio. Fue más importante en mujeres (19,6%) que en hombres (16,5%) y en trabajadores manuales (44,1%) que en no manuales (26,6%). CONCLUSIONES: El contexto en el que los casos de COVID-19 se contagiaron es diferente en función de las desigualdades sociales relativas a país de origen, género y clase social ocupacional.

Human milk inhibits some enveloped virus infections, including SARS-CoV-2, in an intestinal model.

Human milk is important for antimicrobial defense in infants and has well demonstrated antiviral activity. We evaluated the protective ability of human milk against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in a human fetal intestinal cell culture model. We found that, in this model, human milk blocks SARS-CoV-2 replication, irrespective of the presence of SARS-CoV-2 spike-specific antibodies. Complete inhibition of both enveloped Middle East respiratory syndrome coronavirus and human respiratory syncytial virus infections was also observed, whereas no inhibition of non-enveloped enterovirus A71 infection was seen. Transcriptome analysis after 24 h of the intestinal monolayers treated with human milk showed large transcriptomic changes from human milk treatment, and subsequent analysis suggested that <i>ATP1A1</i> down-regulation by milk might be of importance. Inhibition of ATP1A1 blocked SARS-CoV-2 infection in our intestinal model, whereas no effect on EV-A71 infection was seen. Our data indicate that human milk has potent antiviral activity against particular (enveloped) viruses by potentially blocking the ATP1A1-mediated endocytic process.

HDAC Inhibitors Enhance Efficacy of the Oncolytic Adenoviruses Ad∆∆ and Ad-3∆-A20T in Pancreatic and Triple-Negative Breast Cancer Models.

The prognosis for triple-negative breast cancer (TNBC) and pancreatic ductal adenocarcinoma (PDAC) is dismal. TNBC and PDAC are highly aggressive cancers with few treatment options and a potential for rapid resistance to standard-of-care chemotherapeutics. Oncolytic adenoviruses (OAds) represent a promising tumour-selective strategy that can overcome treatment resistance and eliminate cancer cells by lysis and host immune activation. We demonstrate that histone deacetylase inhibitors (HDACi) potently enhanced the cancer-cell killing of our OAds, Ad∆∆ and Ad-3∆-A20T in TNBC and PDAC preclinical models. In the TNBC cell lines MDA-MB-436, SUM159 and CAL51, cell killing, viral uptake and replication were increased when treated with sublethal doses of the Class-I-selective HDACis Scriptaid, Romidepsin and MS-275. The pan-HDACi, TSA efficiently improved OAd efficacy, both in vitro and in SUM159 xenograft models in vivo. Cell killing and Ad∆∆ replication was also significantly increased in five PDAC cell lines when pre-treated with TSA. Efficacy was dependent on treatment time and dose, and on the specific genetic alterations in each cell line. Expression of the cancer specific αvß6-integrin supported higher viral uptake of the integrin-retargeted Ad-3∆-A20T in combination with Scriptaid. In conclusion, we demonstrate that inhibition of specific HDACs is a potential means to enhance OAd activity, supporting clinical translation.

[Peripheral arterial disease and anaemia]. / Anemia en la enfermedad arterial periférica.

Iron deficiency anaemia is highly prevalent worldwide. In the surgical patient, anaemia of any cause implies higher morbidity and mortality in the post-operative period. This is especially important in patients with peripheral artery disease, as they have very high rates of anaemia due to iron deficiency or other causes. In intermittent claudication, anaemia is a predictor of death in the medium term. Patients with critical ischaemia have higher prevalence of anaemia and it is an indicator of amputation and death in the medium term. Specific protocols need to be developed for these patients since the natural history of their disease does not allow for the correction of anaemia before surgery.

Red cell ektacytometry in two patients with chronic hemolytic anemia and three new α-spectrin variants.

Red blood cell (RBC) morphology is, in general, the key diagnostic feature for hereditary spherocytosis (HS) and hereditary elliptocytosis (HE). However, in hereditary pyropoikilocytosis (HPP), the severe clinical form of HE, the morphological diagnosis is difficult due to the presence of a RBC morphological picture characterized by a mixture of elliptocytes, spherocytes, tear-drop cells, and fragmented cells. This difficulty increases in new-borns and/or patients requiring frequent transfusions, making impossible the prediction of the disease course or its severity. Recently, it has been demonstrated that the measurement of osmotic gradient ektacytometry (OGE), using a laser-assisted optical rotational ektacytometer LoRRca (MaxSis, RR Mechatronics), allows a clear differentiation between HS and HE, where the truncated osmoscan curve reflects the inability of the already elliptical cells to deform further under shear stress in the face of hypotonicity. In HPP, however, the RBCs appear to have a significantly decreased ability to maintain deformability in these conditions, and the classical trapezoidal profile of HE is less evident or indistinguishable from HS. Here, two unrelated patients with hereditary hemolytic anemia (HHA) due to HPP and HS, respectively, are described with the joint inheritance of a complex set of five genetic defects. Two of these defects are novel alpha-spectrin gene (SPTA1) variants, one is a microdeletion that removes the entire SPTA1 gene, and two are well-known low-expression polymorphic alleles: α-LELY and α-LEPRA. In the HPP patient (ID1), with many circulating spherocytes, the interactions between the two SPTA1 gene variants may lead, in addition to an elongation defect (elliptocytes), to a loss of membrane stability and vesiculation (spherocytes), and RBCs appear to have a significantly decreased ability to maintain deformability in hypotonic conditions. Due to this, the classical trapezoidal profile of HE may become less evident or indistinguishable from HS. The second patient (ID2) was a classical severe form of HS with the presence of more than 20% of spherocytes and few pincered cells. The severity of clinical manifestation is due to the coinheritance of a microdeletion of chromosome 1 that removes the entire SPTA1 gene with a LEPRA SPTA1 variant in trans. The diagnostic interest of both observations is discussed.

New Mutations in HFE2 and TFR2 Genes Causing Non HFE-Related Hereditary Hemochromatosis.

Hereditary hemochromatosis (HH) is an iron metabolism disease clinically characterized by excessive iron deposition in parenchymal organs such as liver, heart, pancreas, and joints. It is caused by mutations in at least five different genes. HFE hemochromatosis is the most common type of hemochromatosis, while non-HFE related hemochromatosis are rare cases. Here, we describe six new patients of non-HFE related HH from five different families. Two families (Family 1 and 2) have novel nonsense mutations in the HFE2 gene have novel nonsense mutations (p.Arg63Ter and Asp36ThrfsTer96). Three families have mutations in the TFR2 gene, one case has one previously unreported mutation (Family A-p.Asp680Tyr) and two cases have known pathogenic mutations (Family B and D-p.Trp781Ter and p.Gln672Ter respectively). Clinical, biochemical, and genetic data are discussed in all these cases. These rare cases of non-HFE related hereditary hemochromatosis highlight the importance of an earlier molecular diagnosis in a specialized center to prevent serious clinical complications.

Blood Rheological Characterization of ß-Thalassemia Trait and Iron Deficiency Anemia Using Front Microrheometry.

The purpose of this work is to develop a hematocrit-independent method for the detection of beta-thalassemia trait (ß-TT) and iron deficiency anemia (IDA), through the rheological characterization of whole blood samples from different donors. The results obtained herein are the basis for the development of a front microrheometry point-of-care device for the diagnosis and clinical follow-up of ß-TT patients suffering hematological diseases and alterations in the morphology of the red blood cell (RBC). The viscosity is calculated as a function of the mean front velocity by detecting the sample fluid-air interface advancing through a microfluidic channel. Different viscosity curves are obtained for healthy donors, ß-TT and IDA samples. A mathematical model is introduced to compare samples of distinct hematocrit, classifying the viscosity curve patterns with respect to the health condition of blood. The viscosity of the fluid at certain shear rate values varies depending on several RBC factors such as shape and size, hemoglobin (Hb) content, membrane rigidity and hematocrit concentration. Blood and plasma from healthy donors are used as reference. To validate their potential clinical value as a diagnostic tool, the viscosity results are compared to those obtained by the gold-standard method for RBC deformability evaluation, the Laser-Optical Rotational Red Cell Analyzer (LoRRCA).

Parechovirus A Infection of the Intestinal Epithelium: Differences Between Genotypes A1 and A3.

Human parechovirus (PeV-A), one of the species within the Picornaviridae family, is known to cause disease in humans. The most commonly detected genotypes are PeV-A1, associated with mild gastrointestinal disease in young children, and PeV-A3, linked to severe disease with neurological symptoms in neonates. As PeV-A are detectable in stool and nasopharyngeal samples, entry is speculated to occur via the respiratory and gastro-intestinal routes. In this study, we characterized PeV-A1 and PeV-A3 replication and tropism in the intestinal epithelium using a primary 2D model based on human fetal enteroids. This model was permissive to infection with lab-adapted strains and clinical isolates of PeV-A1, but for PeV-A3, infection could only be established with clinical isolates. Replication was highest with infection established from the basolateral side with apical shedding for both genotypes. Compared to PeV-A1, replication kinetics of PeV-A3 were slower. Interestingly, there was a difference in cell tropism with PeV-A1 infecting both Paneth cells and enterocytes, while PeV-A3 infected mainly goblet cells. This difference in cell tropism may explain the difference in replication kinetics and associated disease in humans.