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Rift Valley fever (RVF) is an important zoonotic viral disease affecting several species of domestic and wild ruminants, causing major economic losses and dozens of human deaths in various geographical areas of Africa, where it is endemic. Although it is not present in Europe, there is a risk of its introduction and spread linked to globalisation and climate change. At present, the only measure that could help to prevent the disease is vaccination of flocks in areas at risk of RVF. Available live attenuated vaccines are an effective means of controlling the disease, but their use is often questioned due to residual virulence, particularly in susceptible hosts such as pregnant sheep. On the other hand, no vaccine is currently licensed for use in humans. The development of safe and effective vaccines is therefore a major area of research. In previous studies, we selected under selective mutagenic pressure a highly attenuated RVFV 56/74 virus variant called 40Fp8. This virus showed an extremely attenuated phenotype in both wild-type and immunodeficient A129 (IFNARKO) mice, yet was still able to induce protective immunity after a single inoculation, thus supporting its use as a safe, live attenuated vaccine. To further investigate its safety, in this work we have analysed the attenuation level of 40Fp8 in immunosuppressed mice (A129) when administered by the intranasal route, and compared it with other attenuated RVF viruses that are the basis of vaccines in use or in development. Our results show that 40Fp8 has a much higher attenuated level than these other viruses and confirm its potential as a candidate for safe RVF vaccine development.

IntroductionBluetongue (BT), caused by bluetongue virus (BTV), is an important arthropod-borne livestock disease listed by the World Organization for Animal Health. Live-attenuated and inactivated vaccines have permitted to control BT but they do not simultaneously protect against the myriad of BTV serotypes. Recently, we identified the highly conserved BTV nonstructural protein NS1 and the N-terminal region of NS2 as antigens capable of conferring multiserotype protection against BTV. MethodsHere, we designed Modified Vaccinia Ankara (MVA) viral vectors that expressed BTV-4 proteins VP2 or VP7 along with NS1 and NS2-Nt as well as MVAs that expressed proteins VP2, VP7 or NS1 and NS2-Nt. ResultsImmunization of IFNAR(-/-) mice with two doses of MVA-NS1-2A-NS2-Nt protected mice from BTV-4M infection by the induction of an antigen-specific T cell immune response. Despite rMVA expressing VP7 alone were not protective in the IFNAR(-/-) mouse model, inclusion of VP7 in the vaccine formulation amplified the cell-mediated response induced by NS1 and NS2-Nt. Expression of VP2 elicited protective non-cross-reactive neutralizing antibodies (nAbs) in immunized animals and improved the protection observed in the MVA-NS1-2A-NS2-Nt immunized mice when these three BTV antigens were co-expressed. Moreover, vaccines candidates co-expressing VP2 or VP7 along with NS1 and NS2-Nt provided multiserotype protection. We assessed protective efficacy of both vaccine candidates in sheep against virulent challenge with BTV-4M.DiscussionImmunization with MVA-VP7-NS1-2A-NS2-Nt partially dumped viral replication and clinical disease whereas administration of MVA-VP2-NS1-2A-NS2-Nt promoted a complete protection, preventing viraemia and the pathology produced by BTV infection.

Abstract Riboflavin Transporter Deficiency (RTD) is a rare genetic, childhood-onset disease. This pathology has a relevant neurological involvement, being characterized by motor symptoms, ponto-bulbar paralysis and sensorineural deafness. Such clinical presentation is associated with muscle weakness and motor neuron (MN) degeneration, so that RTD is considered part of the MN disease spectrum. Based on previous findings demonstrating energy dysmetabolism and mitochondrial impairment in RTD induced Pluripotent Stem cells (iPSCs) and iPSC-derived MNs, here we address the involvement of intrinsic apoptotic pathways in disease pathogenesis using these patient-specific in vitro models by combined ultrastructural and confocal analyses. We show impaired neuronal survival of RTD iPSCs and MNs. Focused Ion Beam/Scanning Electron Microscopy (FIB/SEM) documents severe alterations in patients’ cells, including deranged mitochondrial ultrastructure, and altered plasma membrane and nuclear organization. Occurrence of aberrantly activated apoptosis is confirmed by immunofluorescence and TUNEL assays. Overall, our work provides evidence of a role played by mitochondrial dysfunction in RTD, and identifies neuronal apoptosis as a contributing event in disease pathogenesis, indicating intrinsic apoptosis pathways as possible relevant targets for more effective therapeutical approaches.

Abstract Several approaches have been developed to analyze the entry of highly pathogenic viruses. In this study, we report the implementation of a Bimolecular Multicellular Complementation (BiMuC) assay to safely and efficiently monitor SARS-CoV-2 S-mediated membrane fusion without the need for microscopy-based equipment. Using BiMuC, we screened a library of approved drugs and identified compounds that enhance S protein-mediated cell-cell membrane fusion. Among them, ethynylestradiol promotes the growth of SARS-CoV-2 and Influenza A virus in vitro. Our findings demonstrate the potential of BiMuC for identifying small molecules that modulate the life cycle of enveloped viruses, including SARS-CoV-2.

Amyotrophic Lateral Sclerosis (ALS) is considered the prototype of motor neuron disease, characterized by motor neuron loss and muscle waste. A well-established pathogenic hallmark of ALS is mitochondrial failure, leading to bioenergetic deficits. So far, pharmacological interventions for the disease have proven ineffective. Trimetazidine (TMZ) is described as a metabolic modulator acting on different cellular pathways. Its efficacy in enhancing muscular and cardiovascular performance has been widely described, although its molecular target remains elusive. We addressed the molecular mechanisms underlying TMZ action on neuronal experimental paradigms. To this aim, we treated murine SOD1G93A-model-derived primary cultures of cortical and spinal enriched motor neurons, as well as a murine motor-neuron-like cell line overexpressing SOD1G93A, with TMZ. We first characterized the bioenergetic profile of the cell cultures, demonstrating significant mitochondrial dysfunction that is reversed by acute TMZ treatments. We then investigated the effect of TMZ in promoting autophagy processes and its impact on mitochondrial morphology. Finally, we demonstrated the effectiveness of TMZ in terms of the mitochondrial functionality of ALS-rpatient-derived peripheral blood mononuclear cells (PBMCs). In summary, our results emphasize the concept that targeting mitochondrial dysfunction may represent an effective therapeutic strategy for ALS. The findings demonstrate that TMZ enhances mitochondrial performance in motor neuron cells by activating autophagy processes, particularly mitophagy. Although further investigations are needed to elucidate the precise molecular pathways involved, these results hold critical implications for the development of more effective and specific derivatives of TMZ for ALS treatment.

Vitamin B12 (VitB12) is a micronutrient and acts as a cofactor for fundamental biochemical reactions: the synthesis of succinyl-CoA from methylmalonyl-CoA and biotin, and the synthesis of methionine from folic acid and homocysteine. VitB12 deficiency can determine a wide range of diseases, including nervous system impairments. Although clinical evidence shows a direct role of VitB12 in neuronal homeostasis, the molecular mechanisms are yet to be characterized in depth. Earlier investigations focused on exploring the biochemical shifts resulting from a deficiency in the function of VitB12 as a coenzyme, while more recent studies propose a broader mechanism, encompassing changes at the molecular/cellular levels. Here, we explore existing study models employed to investigate the role of VitB12 in the nervous system, including the challenges inherent in replicating deficiency/supplementation in experimental settings. Moreover, we discuss the potential biochemical alterations and ensuing mechanisms that might be modified at the molecular/cellular level (such as epigenetic modifications or changes in lysosomal activity). We also address the role of VitB12 deficiency in initiating processes that contribute to nervous system deterioration, including ROS accumulation, inflammation, and demyelination. Consequently, a complex biological landscape emerges, requiring further investigative efforts to grasp the intricacies involved and identify potential therapeutic targets.

The introduction of three single nucleotide mutations into the genome of the virulent RVFV ZH548 strain allows for the rescue of a fully attenuated virus in mice (ZH548-rA2). These mutations are located in the viral genes encoding the RdRp and the non-structural protein NSs. This paper shows the results obtained after the subcutaneous inoculation of ZH548-rA2 in adult sheep and the subsequent challenge with the parental virus (ZH548-rC1). Inoculation with the ZH548-rA2 virus caused no detectable clinical or pathological effect in sheep, whereas inoculation of the parental rC1 virus caused lesions compatible with viral infection characterised by the presence of scattered hepatic necrosis. Viral infection was confirmed via immunohistochemistry, with hepatocytes within the necrotic foci appearing as the main cells immunolabelled against viral antigen. Furthermore, the inoculation of sheep with the rA2 virus prevented the liver damage expected after rC1 virus inoculation, suggesting a protective efficacy in sheep which correlated with the induction of both humoral and cell-mediated immune responses.

Abstract To define the seroprevalence of antibodies against SARS-CoV-2 in the municipality of Vilanova del Camí (in the region of Conca d'Ódena, Barcelona, Spain) and to know the risk factors associated with positive seroprevalence. Cross-sectional descriptive study. The population of Vilanova del Camí had the opportunity to voluntarily attend two screenings (October and December 2020) for antibodies against the nucleocapsid protein of SARS-CoV-2 using a Rapid Diagnostic Test (RDT) (Salocor (Salofa Oy). Participants in the screening signed an informed consent form. From the 3,610 attendees at the screening, 2,170 patients were randomly selected. The relationship between antibody test results and other demographic (sex, age, morbidity index) and clinical (diagnoses, smoking and drugs) variables was analysed. The prevalence of antibodies against SARS-CoV-2 was 9.6% (95% CI of 8.4% to 10.9%) and was similar for men and women but increased with age. Among complex chronic patients, 14.3% had antibodies against SARS-CoV-2, and among patients with advanced chronic disease, 25% had antibodies against SARS-CoV-2. Age, AMG (Adjusted Morbidity Groups) index, COVID-19 diagnosis and contact with a COVID-19 case were risk factors for positive seroprevalence. A higher seroprevalence was detected in the October screening (12.16%) than in the December screening (8.38%). In the December screening, obesity was a risk factor for positive seroprevalence. This study demonstrates the high seroprevalence of antibodies against SARS-CoV-2 in the pandemic epicentre of Catalonia.

Abstract Cold Atmospheric Plasma (CAP) and Plasma Activated Media (PAM) are effective against bacteria, fungi, cancer cells, and viruses because they can deliver Reactive Oxygen and Nitrogen Species (RONS) on a living tissue with negligible damage on health cells. The antiviral activity of CAP against SARS-CoV-2 is being investigated, however, the same but of PAM has not been explored despite its potential. In the present study, the capability of Plasma Activated Media (PAM) to inactivate SARS-CoV-2 and PR8 H1N1 influenza virus with negligible damage on healthy cells is demonstrated. PAM acted by both virus detaching and diminished replication. Furthermore, the treatment of A549 lung cells at different times with buffered PAM did not induce interleukin 8 expression, showing that PAM did not induce inflammation. These results open a new research field by using PAM to the development novel treatments for COVID-19, influenza, and other respiratory diseases.

IntroductionCoronavirus disease 2019 (COVID-19) is an infectious zoonotic disease caused by SARS-CoV-2. Monitoring the infection in pets is recommended for human disease surveillance, prevention, and control since the virus can spread from people to animals during close contact. Several diagnostic tests have been adapted from humans to animals, but limited data on the validation process are available.MethodsHerein, the first comparative study of six “in house” and two commercial serological tests developed to monitor SARS-CoV-2 infection in pets was performed with a well-coded panel of sera (61 cat sera and 74 dog sera) with a conservative criterion (viral seroneutralisation and/or RT–qPCR results) as a reference. Four “in house” tests based on either the RBD fragment of the spike protein (RBD-S) or the N-terminal fragment of the nucleoprotein (N) were developed for the first time. The analytical specificity (ASp) of those tests that showed the best diagnostic performance was assessed. The validation included the analysis of a panel of sera obtained pre-pandemic from cats and dogs infected with other coronaviruses to determine the analytical Sp (17 cat sera and 41 dog sera).Results and discussionELISAS based on the S protein are recommended in serosurveillance studies for cats (RBD-S SALUVET ELISA, ELISA COVID UNIZAR and INgezim® COVID 19 S VET) and dogs (INgezim® COVID 19 S VET and RBD-S SALUVET ELISA). These tests showed higher diagnostic sensitivity (Se) and DSp in cats (>90%) than in dogs. When sera obtained prior to the pandemic and from animals infected with other coronaviruses were analyzed by RBD-S and N SALUVET ELISAs and INgezim® COVID 19 S VET, a few cross reactors or no cross reactions were detected when dog and cat sera were analyzed by tests based on the S protein, respectively. In contrast, the number of cross reactions increased when the test was based on the N protein. Thus, the use of tests based on the N protein was discarded for serodiagnosis purposes. The results obtained revealed the most accurate serological tests for each species. Further studies should attempt to improve the diagnostic performance of serological tests developed for dogs.

Resumen: Objetivo: Evaluar las fuentes de información empleadas por los Registros Autonómicos de Enfermedades Raras (RAER) para la captación de la enfermedad de Wilson en España, calcular su prevalencia y mortalidad, y describir las características sociodemográficas de las personas afectadas. Método: Estudio epidemiológico transversal, periodo 2010-2015. Se captaron los posibles casos mediante los códigos 275.1 (CIE-9-MC), E83.0 (CIE-10) y 905 ORPHA en 15 RAER y el Registro de Pacientes de Enfermedades Raras del Instituto de Salud Carlos III. Los diagnósticos fueron validados revisando la documentación clínica. Se calcularon el valor predictivo positivo (VPP) de las fuentes de información, la prevalencia, la mortalidad y la distribución de las características sociodemográficas. Resultados: El Conjunto Mínimo Básico de Datos (CMBD) fue la fuente de información más utilizada por los RAER (VPP = 39,4%), seguida del Registro de Medicamentos Huérfanos (RMH) (VPP = 81,9%). La Historia Clínica de Atención Primaria (HCAP) obtuvo un VPP del 55,9%. Las combinaciones con mayor VPP fueron las del RMH con el CMBD (VPP = 95,8%) y del RMH con la HCAP (VPP = 92,9%). Se confirmaron 514 casos, el 57,2% eran hombres, cuya edad mediana de diagnóstico fue de 21,3 años. La prevalencia fue de 1,64/100.000 habitantes en 2015 y la mortalidad del 3,0%, siendo ambas superiores en los hombres. Conclusión: Se recomienda la incorporación en los RAER del RMH y de la HCAP, ya que su combinación y la del RMH con el CMBD podrían utilizarse como criterio de validación automática para la enfermedad de Wilson. La prevalencia obtenida fue similar a la de otros países próximos a España. Abstract: Objective: To evaluate the sources of information used by the Regional Population-based Registries of Rare Diseases (RRD) for Wilson's Disease identification in Spain; to calculate its prevalence and mortality; and to describe the sociodemographic characteristics of those affected. Method: Cross-sectional epidemiological study, period 2010-2015. Possible cases were identified by codes 275.1 (ICD-9-CM), E83.0 (ICD-10) and 905 (ORPHAcode) in: 15 participating RRD and the Rare Disease Patients Registry of the Carlos III Health Institute. The diagnoses were confirmed through a clinical documentation review. The positive predictive value (PPV) of the sources of information used by RRD and their combinations were obtained. The prevalence, mortality and the distribution of sociodemographic characteristics were calculated. Results: The Hospital Discharge Database (HDD) was the most used source by the RRD (PPV = 39.4%), followed by the Orphan Drugs Registry (ODR) (PPV = 81.9%). The Clinical History of Primary Care (PC) obtains PPV = 55.9%. The combinations with highest PPV were the ODR with HDD (PPV = 95.8%) and the ODR with PC (PPV = 92.9%). 514 cases were confirmed, 57.2% men, with a median age of diagnosis of 21.3 years. The prevalence was 1.64/100,000 inhabitants in 2015 and mortality rate was 3.0%, being both higher in men. Conclusions: Incorporation of ODR and PC into the RRD is recommended, as its combination and ODR with HDD could be used as an automatic validation criterion for Wilson's disease. The prevalence obtained was similar to that of countries close to Spain.

Controlled fermentation processes have high potential for improving coffee quality. The effect of fermentation temperature on beverage quality was investigated with coffee cultivated at elevations between 1166 and 1928 m. A completely randomized design was carried out at five elevation ranges at 200 m intervals in five farms per elevation range, and two temperatures (15 °C and 30 °C), which were maintained in a temperature-controlled bioreactor. Each temperature-controlled fermentation batch had a spontaneous fermentation batch (control treatment). Microbial identification of LAB and yeast was performed using a Biolog™ MicroStation™ ID System, and cup quality tests were performed following the SCA protocol. Tests conducted at 15 °C showed higher microbial community activity on the substrates used, indicating greater transformation potential than those conducted at 30 °C or those of spontaneous fermentation. According to Wilcoxon and Kruskal–Wallis tests, temperature-controlled fermentation resulted in high-quality coffee for all elevation ranges, with coffee from higher elevations and processed at controlled temperatures of 15 °C receiving the highest cup scores compared to coffee that was subjected to 30 °C. These results suggest that controlled temperature can be used to design standardized fermentation processes in order to enhance coffee quality through differentiated sensory profiles.

The COVID-19 pandemic has underscored the importance of swift responses and the necessity of dependable technologies for vaccine development. Our team previously developed a fast cloning system for the modified vaccinia virus Ankara (MVA) vaccine platform. In this study, we reported on the construction and preclinical testing of a recombinant MVA vaccine obtained using this system. We obtained recombinant MVA expressing the unmodified full-length SARS-CoV-2 spike (S) protein containing the D614G amino-acid substitution (MVA-Sdg) and a version expressing a modified S protein containing amino-acid substitutions designed to stabilize the protein a in a pre-fusion conformation (MVA-Spf). S protein expressed by MVA-Sdg was found to be expressed and was correctly processed and transported to the cell surface, where it efficiently produced cell–cell fusion. Version Spf, however, was not proteolytically processed, and despite being transported to the plasma membrane, it failed to induce cell–cell fusion. We assessed both vaccine candidates in prime-boost regimens in the susceptible transgenic K18-human angiotensin-converting enzyme 2 (K18-hACE2) in mice and in golden Syrian hamsters. Robust immunity and protection from disease was induced with either vaccine in both animal models. Remarkably, the MVA-Spf vaccine candidate produced higher levels of antibodies, a stronger T cell response, and a higher degree of protection from challenge. In addition, the level of SARS-CoV-2 in the brain of MVA-Spf inoculated mice was decreased to undetectable levels. Those results add to our current experience and range of vaccine vectors and technologies for developing a safe and effective COVID-19 vaccine.

Abstract The optical absorption coefficient of culture media is critical for the survival analysis of pathogens under optical irradiation. The quality of the results obtained from experiments relies on the optical analysis of the spatial distribution of fluence which also depends on the geometry of the sample. In this contribution, we consider both the geometrical shape and the culture medium’s absorption coefficient to evaluate how the spatial distribution of optical radiation affects pathogens/viruses. In this work, we exposed SARS-CoV-2 to UV-C radiation ( $$\lambda$$ λ = 254 nm) and we calculated—considering the influence of the optical absorption of the culture medium—a characteristic inactivation fluence of $$F_i$$ F i = 4.7 J/m2, or an equivalent 10% survival (D90 dose) of 10.8 J/m2. Experimentally, we diluted the virus into sessile drops of Dulbecco’s Modified Eagle Medium to evaluate pathogen activity after controlled doses of UV irradiation. To validate the optical absorption mode, we carried out an additional experiment where we varied droplet size. Our model—including optical absorption and geometrical considerations—provides robust results among a variety of experimental situations, and represents our experimental conditions more accurately. These results will help to evaluate the capability of UV disinfecting strategies applied to a variety of everyday situations, including the case of micro-droplets generated by respiratory functions.

BET proteins function as histone code readers of acetylated lysins that determine the positive regulation in transcription of genes involved in cell cycle progression, differentiation, inflammation, and many other pathways. In recent years, thanks to the development of BET inhibitors, interest in this protein family has risen for its relevance in brain development and function. For example, experimental evidence has shown that BET modulation affects neuronal activity and the expression of genes involved in learning and memory. In addition, BET inhibition strongly suppresses molecular pathways related to neuroinflammation. These observations suggest that BET modulation may play a critical role in the onset and during the development of diverse neurodegenerative and neuropsychiatric disorders, such as Alzheimer’s disease, fragile X syndrome, and Rett syndrome. In this review article, we summarize the most recent evidence regarding the involvement of BET proteins in brain physiology and pathology, as well as their pharmacological potential as targets for therapeutic purposes.

Telomerase reverse transcriptase (TERT) is the catalytic subunit of telomerase holoenzyme, which adds telomeric DNA repeats on chromosome ends to counteract telomere shortening. In addition, there is evidence of TERT non-canonical functions, among which is an antioxidant role. In order to better investigate this role, we tested the response to X-rays and H2O2 treatment in hTERT-overexpressing human fibroblasts (HF-TERT). We observed in HF-TERT a reduced induction of reactive oxygen species and an increased expression of the proteins involved in the antioxidant defense. Therefore, we also tested a possible role of TERT inside mitochondria. We confirmed TERT mitochondrial localization, which increases after oxidative stress (OS) induced by H2O2 treatment. We next evaluated some mitochondrial markers. The basal mitochondria quantity appeared reduced in HF-TERT compared to normal fibroblasts and an additional reduction was observed after OS; nevertheless, the mitochondrial membrane potential and morphology were better conserved in HF-TERT. Our results suggest a protective function of TERT against OS, also preserving mitochondrial functionality.

Background: the incidence of lung cancer in the province of Sancti Spíritus is high and its diagnosis is usually made in advanced stages, for which surgical treatment is not always possible. Objective: to analyze the main characteristics and results of patients operated on for lung cancer. Methods: a cross-sectional descriptive observational study was carried out in the General Surgery service of the Camilo Cienfuegos General University Hospital in the Sancti Spiritus province in the period between January 1996 and December 2019. The universe consisted of 83 patients operated with lung cancer who underwent resection accompanied by mediastinal lymph node sampling. Results: the male sex predominated in the studied group. The most frequent age groups for both sexes were those from 51 to 60 years old and those over 60 years old. Smoking was present in most of the patients. The most frequent tumor location was in the upper lobes and lobectomy was the most performed technique. The pathological stages that predominated were lB and llB, followed by lllA. Squamous cell carcinoma was the predominant histological type. Atelectasis and respiratory infections were the most frequent complications. The deceased were few. Conclusions: the demographic characteristics of the patients studied correspond to other series of patients with lung cancer. Tumor location in the upper lobes, squamous cell carcinoma, as well as early stages, predominated in the series studied. Lobectomy was the most frequently used surgical technique and among complications, respiratory ones were the most frequent. DeCS: LUNG NEOPLASMS/surgery; LUNG NEOPLASMS/diagnosis; LUNG NEOPLASMS/complications; PNEUMONECTOMY/methods; CARCINOMA, SQUAMOUS CELL/complications.

Abstract Schmallenberg virus (SBV), an arthropod-transmitted pathogenic bunyavirus, continues to be a threat to the European livestock industry, causing morbidity and mortality among young ruminant livestock. Here, we describe a novel SBV subunit vaccine, based on bacterially expressed SBV nucleoprotein (SBV-N) administered with a veterinary-grade Saponin adjuvant. When assayed in an IFNAR-/- mouse model, SBV-N with Saponin induced strong non-neutralizing broadly virus-reactive antibodies, decreased clinical signs, as well as significantly reduced viremia. Vaccination assays also suggest that this level of immune protection is cell mediated, as evidenced by the lack of neutralizing antibodies, as well as interferon-γ secretion observed in vitro. Therefore, based on these results, bacterially expressed SBV-N, co-administered with veterinary-grade Saponin adjuvant may serve as a promising economical alternative to current SBV vaccines, and warrant further evaluation in large ruminant animal models. Moreover, we propose that this strategy may be applicable to other bunyaviruses.

Rift Valley fever (RVF) is an arboviral zoonotic disease affecting many African countries with the potential to spread to other geographical areas. RVF affects sheep, goats, cattle and camels, causing a high rate of abortions and death of newborn lambs. Also, humans can be infected, developing a usually self-limiting disease that can turn into a more severe illness in a low percentage of cases. Although different veterinary vaccines are available in endemic areas in Africa, to date no human vaccine has been licensed. In previous works, we described the selection and characterization of a favipiravir-mutagenized RVFV variant, termed 40Fp8, with potential as a RVF vaccine candidate due to the strong attenuation shown in immunocompromised animal models. Compared to the parental South African 56/74 viral strain, 40Fp8 displayed 7 amino acid substitutions in the L-protein, three of them located in the central region corresponding to the catalytic core of the RNA-dependent RNA polymerase (RdRp). In this work, by means of a reverse genetics system, we have analyzed the effect on virulence of these amino acid changes, alone or combined, both in vitro and in vivo. We found that the simultaneous introduction of two changes (G924S and A1303T) in the heterologous ZH548-RVFV Egyptian strain conferred attenuated phenotypes to the rescued viruses as shown in infected mice without affecting virus immunogenicity. Our results suggest that both changes induce resistance to favipiravir likely associated to some fitness cost that could be the basis for the observed attenuation in vivo. Conversely, the third change, I1050V, appears to be a compensatory mutation increasing viral fitness. Altogether, these results provide relevant information for the safety improvement of novel live attenuated RVFV vaccines.

A systematic review of the literature in PubMed was made by combining the terms “cuspid” and “tooth root” as MeSH health descriptors, combined with the Boolean operators “+” and “&” to obtain describing publications about two roots canines in order to sustain, on scientific evidence, the application of dental anthropology (dental morphological variations) in forensic science (forensic processes of dental identification). This literature review identified reports that describe the presence of two-rooted canines and the number and distribution of root canals for diagnostic and therapeutic purposes; and one report in which description was performed for forensic identification purposes. The descriptions corresponded to different cases of permanent maxillary canines with left unilateral expression, permanent mandibular canines with right unilateral expression, left unilateral expression with bilateral expression. There were no reports of deciduous dentition. Likewise, a case report in which skeletonized human remains were identified by the presence of bilateral two-root mandibular deciduous canines is described.

Background: Murine leprosy is a chronic granulomatous disease caused by Mycobacterium lepraemurium (MLM) in mice and rats. The disease evolves with the development of cellular anergy that impedes the production of interferon gamma (IFNγ), tumor necrosis factor-alpha (TNFα), and nitric oxide (NO) required to kill the microorganism. In this study we investigated whether histone deacetylase inhibitors (HDACi) (valproic acid and sodium butyrate [NaB]) and the immunomodulator transfer factor in dialyzable leukocyte extracts (DLE) can prevent anergy in murine leprosy. Methods: Five groups of six Balb/c mice were intraperitoneally inoculated with 2 × 107 MLM. Thirty-days post inoculation, treatment was started; one group received no treatment, one was treated with rifampicin-clofazimine (R-C), one with sodium valproate (VPA), one with NaB, and one with DLE. The animals were monitored for the evidence of disease for 96 days. After euthanasia, their spleens were removed and processed for histologic, bacteriologic, and cytokine studies. Results: R-C completely controlled the ongoing disease. DLE and NaB significantly reduced the development of lesions, including granuloma size and the number of bacilli; VPA was less effective. DLE, NaB, and VPA reverted the anergic condition in diverse grades and allowed the expression of IFNγ, TNFα, and inducible NO synthase, also in diverse grades. Conclusion: Anergy in leprosy and murine leprosy allows disease progression. In this study, anergy was prevented, in significant degree, by DLE (an immunomodulator) and NaB (HDACi). VPA was less effective. These results suggest potential beneficial effects of DLE and NaB in the ancillary treatment of leprosy.

Inorganic materials can provide a set of tools to decontaminate solid, liquid or air containing viral particles. The use of disinfectants can be limited or not practical in scenarios where continuous cleaning is not feasible. Physicochemical differences between viruses raise the need for effective formulations for all kind of viruses. In the present work we describe two types of antimicrobial inorganic materials: i) a novel soda-lime glass (G3), and ii) kaolin containing metals nanoparticles (Ag or CuO), as materials to disable virus infectivity. Strong antiviral properties can be observed in G3 glass, and kaolin-containing nanoparticle materials showing a reduction of viral infectivity close to 99% in the first 10 ​min of contact of vesicular stomatitis virus (VSV). A potent virucidal activity is also present in G3 and kaolin containing Ag or CuO nanoparticles against all kinds of viruses tested, reducing more than 99% the amount of HSV-1, Adenovirus, VSV, Influenza virus and SARS-CoV-2 exposed to them. Virucidal properties could be explained by a direct interaction of materials with viruses as well as inactivation by the presence of virucidal elements in the material lixiviates. Kaolin-based materials guarantee a controlled release of active nanoparticles with antiviral activity. Current coronavirus crisis highlights the need for new strategies to remove viruses from contaminated areas. We propose these low-cost inorganic materials as useful disinfecting antivirals in the actual or future pandemic threats.

Breast cancer is the first leading tumor in women in terms of incidence worldwide. Seventy percent of cases are estrogen receptor (ER) α-positive. In these malignancies, 17β-estradiol (E2) via ERα increases the levels of neuroglobin (NGB), a compensatory protein that protects cancer cells from stress-induced apoptosis, including chemotherapeutic drug treatment. Our previous data indicate that resveratrol (RSV), a plant-derived polyphenol, prevents E2/ERα-induced NGB accumulation in this cellular context, making E2-dependent breast cancer cells more prone to apoptosis. Unfortunately, RSV is readily metabolized, thus preventing its effectiveness. Here, four different RSV analogs have been developed, and their effect on the ERα/NGB pathway has been compared with RSV conjugated with highly hydrophilic gold nanoparticles as prodrug to evaluate if RSV derivatives maintain the breast cancer cells’ susceptibility to the chemotherapeutic drug paclitaxel as the original compound. Results demonstrate that RSV conjugation with gold nanoparticles increases RSV efficacy, with respect to RSV analogues, reducing NGB levels and enhancing the pro-apoptotic action of paclitaxel, even preventing the anti-apoptotic action exerted by E2 treatment on these cells. Overall, RSV conjugation with gold nanoparticles makes this complex a promising agent for medical application in breast cancer treatment.

Frente a las realidades imperantes que agobian y perturban la vida de la gente, constituye un compromiso imprescindible educar para una vida digna. Y pensar en ello, es también pensar en Paulo Freire y su educación liberadora, pues educar para una vida digna, exige que se favorezca la posibilidad de ser y desplegar todas las cualidades humanas en la búsqueda de transitar hacia su ideal de vida buena, lograr condiciones sociales y culturales necesarias para garantizar bienestar y una organización social justa para las presentes y futuras generaciones de todos los sectores de nuestras sociedades plurales. Y ante esta urgencia, en diálogo con el pensamiento freireano, nos aproximarnos a algunos escenarios que, desde nuestra perspectiva, se constituyen en espacios que confluyen como claves para dotar de un sentido – otro al acto de educar, como posibilitad de educar para una vida digna, como apuesta trasformadora. Es así como asumimos cinco claves como horizontes para pensar la educación del presente como vía para transitar el espacio de lo posible por–venir: educar para la ética y la política, educar para la diferencia, educar para el amor, educar para la resistencia y la liberación, educar para la experiencia.

Riboflavin transporter deficiency (RTD) is a rare genetic disorder characterized by motor, sensory and cranial neuropathy. This childhood-onset neurodegenerative disease is caused by biallelic pathogenic variants in either SLC52A2 or SLC52A3 genes, resulting in insufficient supply of riboflavin (vitamin B2) and consequent impairment of flavoprotein-dependent metabolic pathways. Current therapy, empirically based high-dose riboflavin supplementation, ameliorates the progression of the disease, even though response to treatment is variable and partial. Recent studies have highlighted concurrent pathogenic contribution of cellular energy dysmetabolism and cytoskeletal derangement. In this context, patient specific RTD models, based on induced pluripotent stem cell (iPSC) technology, have provided evidence of redox imbalance, involving mitochondrial and peroxisomal dysfunction. Such oxidative stress condition likely causes cytoskeletal perturbation, associated with impaired differentiation of RTD motor neurons. In this review, we discuss the most recent findings obtained using different RTD models. Relevantly, the integration of data from innovative iPSC-derived in vitro models and invertebrate in vivo models may provide essential information on RTD pathophysiology. Such novel insights are expected to suggest custom therapeutic strategies, especially for those patients unresponsive to high-dose riboflavin treatments.

The cytoskeletal network plays a crucial role in the differentiation, morphogenesis, function and homeostasis of the nervous tissue, so that alterations in any of its components may lead to neurodegenerative diseases. Riboflavin transporter deficiency (RTD), a childhood-onset disorder characterized by degeneration of motor neurons (MNs), is caused by biallelic mutations in genes encoding the human riboflavin (RF) transporters. In a patient-specific induced pluripotent stem cells (iPSCs) model of RTD, we recently demonstrated altered cell-cell contacts, energy dysmetabolism and redox imbalance. The present study focuses on cytoskeletal composition and dynamics associated to RTD, utilizing patients' iPSCs and derived MNs. Abnormal expression and distribution of α- and β-tubulin (α- and β-TUB), as well as imbalanced tyrosination of α-TUB, accompanied by an impaired ability to re-polymerize after nocodazole treatment, were found in RTD patient-derived iPSCs. Following differentiation, MNs showed consistent changes in TUB content, which was associated with abnormal morphofunctional features, such as neurite length and Ca2+ homeostasis, suggesting impaired differentiation. Beneficial effects of RF supplementation, alone or in combination with the antioxidant molecule N-acetyl cystine (NAC), were assessed. RF administration resulted in partially improved cytoskeletal features in patients' iPSCs and MNs, suggesting that redundancy of transporters may rescue cell functionality in the presence of adequate concentrations of the vitamin. Moreover, supplementation with NAC was demonstrated to be effective in restoring all the considered parameters, when used in combination with RF, thus supporting the therapeutic use of both compounds.

Plant endophytic bacteria have received special attention in recent decades for their ability to improve plant response to multiple stresses. A positive effect of endophytes on plant’s ability to cope with drought, salinity, nitrogen deficiency, and pathogens have already been demonstrated in numerous studies, and recently this evidence was consolidated in a meta-analysis of published data. Endophytic bacteria have also been implicated in increasing resistance to heavy metals in plants; despite the important biotechnological applications of such effect in heavy metal bioremediation and agriculture, efforts to systematically analyze studies in this field have been limited. In this study, we address this task with the objective of establishing whether the findings made for other types of stresses extend to the response to heavy metals. Specifically, we seek to establish if plant inoculation with plant-growth promoting endophytic bacteria have an impact on their tolerance to heavy metal stress? We carried out a meta-analysis of the effect size of inoculation with endophytic bacteria on the host plant biomass in response to heavy metal stress (aluminum, arsenic, cadmium, copper, chromium, manganese, nickel, lead, and zinc), which included 27 (from 76 published in the last 10 years) studies under controlled conditions that evaluated 19 host plants and 20 bacterial genera. Our results suggest that endophytic bacteria increase the biomass production of host plants subjected to different heavy metals, indicating their effectiveness in protecting plants from a wide range of metal toxicities. Stress mitigation by the bacteria was similar among the different plant groups with the exception of non-accumulating plants that benefit most from the symbiotic association. Host identity and heavy metal concentration seem to influence the effect of the bacteria. Our analysis revealed that bacterial consortia provide the greatest benefit although the most common biotechnological applications are not directed towards them, and support the value of endophytic bacteria as an alternative to mitigate heavy metal stress in a wide variety of hosts.

Este es un estudio descriptivo transversal cuantitativo que identificó el modo de herencia de 14 Rasgos Morfológicos Dentales (RMDC) a partir de la frecuencia, la variablidad, el dimorfismo sexual, la asimetría bilateral y la correspondencia entre rasgos, en modelos de yeso obtenidos de un grupo de diez familias —madre, padre e hijo(a)— que forman parte de la comunidad misak del municipio de Silvia (Cauca, Colombia). Se emplearon los sistemas de registro para dentición decidua Arizona State University Dental Anthropology System (ASUDAS, 1991) para crowding, winging y cúspide 6, Grine (1986) para cúspide de Carabelli, Sciuli (1998) para protostílido, pliegue acodado, patrón cuspídeo y número de cúspides, y Hanihara (1966) para cúspide 7; y para dentición permanente ASUDAS para los 14 RMDC. Se observaron altas frecuencias de punto P del protostílido, pliegue acodado, patrón cuspídeo, número de cúspides y cúspide 6 en los dientes deciduos; y de crowding, winging, forma de pala, reducción de hipocono, punto P del protostílido, patrón cuspídeo y cúspide 6 en los dientes permanentes.No hubo dimorfismo sexual y se observó simetría bilateral. Hubo correspondencia entre la dentición decidua y permanente para la expresión de la cúspide de Carabelli, el número de cúspides y la cúspide 6. Se observó una tendencia de herencia de la expresión dicotómica presencia/ausencia de los RMDC desde las madres hacia los hijos. El grupo de indígenas Misak corresponde a una población con alta influencia del complejo dental mongoloide y baja influencia de los complejos dentales caucasoide y negroide de acuerdo con la expresión y variabilidad de los RMDC. La ausencia de dimorfismo sexual, la presencia de simetría bilateral y la correspondencia de algunos rasgos, evidencia un fuerte control genético en el modo de herencia (con tendencia desde las madres hacia los hijos) y una baja influencia del medio ambiente.

SARS-CoV-2 infection in hospital areas is of a particular concern, since the close interaction between health care personnel and patients diagnosed with COVID-19, which allows virus to be easily spread between them and subsequently to their families and communities. Preventing SARS-CoV-2 infection among healthcare personnel is essential to reduce the frequency of infections and outbreaks during the pandemic considering that they work in high-risk areas. In this research, silver nanoparticles (AgNPs) were tested in vitro and shown to have an inhibitory effect on SARS-CoV-2 infection in cultured cells. Subsequently, we assess the effects of mouthwash and nose rinse with ARGOVIT® silver nanoparticles (AgNPs), in the prevention of SARS-CoV-2 contagion in health workers consider as high-risk group of acquiring the infection in the General Tijuana Hospital, Mexico, a hospital for the exclusive recruitment of patients diagnosed with COVID-19. We present a prospective randomized study of 231 participants that was carried out for 9 weeks (during the declaration of a pandemic). The "experimental" group was instructed to do mouthwash and nose rinse with the AgNPs solution; the "control" group was instructed to do mouthwashes and nose rinse in a conventional way. The incidence of SARS-CoV-2 infection was significantly lower in the "experimental" group (two participants of 114, 1.8%) compared to the "control" group (thirty-three participants of 117, 28.2%), with an 84.8% efficiency. We conclude that the mouth and nasal rinse with AgNPs helps in the prevention of SARS-CoV-2 infection in health personnel who are exposed to patients diagnosed with COVID-19.

BackgroundMany ruminant diseases of viral aetiology can be effectively prevented using appropriate vaccination measures. For diseases such as Rift Valley fever (RVF) the long inter-epizootic periods make routine vaccination programs unfeasible. Coupling RVF prophylaxis with seasonal vaccination programmes by means of multivalent vaccine platforms would help to reduce the risk of new RVF outbreaks.Methodology/principal findingsIn this work we generated recombinant attenuated Rift Valley fever viruses (RVFVs) encoding in place of the virulence factor NSs either the VP2 capsid protein or a truncated form of the non-structural NS1 protein of bluetongue virus serotype 4 (BTV-4). The recombinant viruses were able to carry and express the heterologous BTV genes upon consecutive passages in cell cultures. In murine models, a single immunization was sufficient to protect mice upon RVFV challenge and to elicit a specific immune response against BTV-4 antigens that was fully protective after a BTV-4 boost. In sheep, a natural host for RVFV and BTV, both vaccines proved immunogenic although conferred only partial protection after a virulent BTV-4 reassortant Morocco strain challenge.Conclusions/significanceThough additional optimization will be needed to improve the efficacy data against BTV in sheep, our findings warrant further developments of attenuated RVFV as a dual vaccine platform carrying heterologous immune relevant antigens for ruminant diseases in RVF risk areas.

Biochar is a solid material obtained from the thermal decomposition of biomass of diverse biological origins through a process called pyrolysis. Biochar has great potential for reducing greenhouse gas emissions, sequester carbon in the soil, rehabilitate degraded soils, and reduce dependence on chemical fertilizers in crops. It also improves the physical, chemical, and biological properties of the soil and has a positive effect on plant growth. Given these attributes, there is a growing interest for adopting its use in agriculture, soil and land reclamation, and climate change mitigation. The effects of biochar application can be neutral or positive and will be determined mainly by factors such as the origin of the raw materials, carbonization conditions, frequency of applications, the method of application and dosage. In this review, we offer a detailed examination of the origins of biochar and the technologies used for its production. We examine the various materials that have been used to produce biochars and how they affect their physico-chemical characteristics, and we describe their applications in agriculture and climate change mitigation. Finally, we list the guides that describe the standards for the production, characterization, and commercialization of biochar that seek to guarantee the quality of the product and the essential characteristics for its safe use.

Introduction: the cusp pattern and the deflecting wrinkle are two morphological traits whose expression and variability contribute to the morphological configuration of the occlusal surface of deciduous and permanent lower molar teeth. The objective of the present study was to estimate the correlation between cusp pattern and deflecting wrinkle in six ethnic groups from southwestern Colombia. Methods: analysis of 480 plaster models corresponding to 100 mixed Caucasoid mestizos from Cali (50 men and 50 women), 60 Afro-descendants from Cali (34 women and 26 men), 84 Afro-descendants from Puerto Tejada (42 women and 42 men), 116 Afro-descendants from Villa Rica (57 men and 59 women), 60 misak indigenous (37 women and 23 men) and 60 nasa indigenous (35 women and 25 men). Results: there were high frequencies (over 60%) of cusp pattern (Y expression) and deflecting wrinkle (grades 2 and 3) in deciduous first lower molars, mid frequencies (between 40 and 59%) of cusp pattern (Y and + expressions) and deflecting wrinkle (grades 1 and 2) in permanent first lower molars, and mid frequencies (between 40 and 59%) of cusp pattern (+ and X expressions) and deflecting wrinkle (grades 1 and 2) in permanent lower second molars. Conclusions: these frequencies suggest a trend in cusp pattern, which may vary according to the degree of expression of the deflecting wrinkle. This tendency consists of the Y cusp pattern when the deflecting wrinkle is expressed in grades 2 and 3. Bilaterally was observed and there was no sexual dimorphism.

Introducción. La periodontitis es una enfermedad infecciosa que afecta los tejidos de soporte del diente y se asocia con diferentes enfermedades sistémicas, incluida la enfermedad cardiovascular. Los estudios microbiológicos permiten detectar microorganismos a partir de muestras subgingivales y cardiovasculares. Objetivo. Describir la microbiota periodontal cultivable y la presencia de microorganismos en válvulas cardiacas de pacientes sometidos a cirugía de reemplazo valvular en una clínica de Cali. Materiales y métodos. Se analizaron 30 muestras subgingivales y de tejidos valvulares mediante cultivo en medio bifásico, agar de sangre con suplemento y agar tripticasa de soya con antibiótico. Las muestras de las válvulas se analizaron mediante reacción en cadena de la polimerasa (PCR) convencional. Resultados. Los patógenos periodontales aislados de bolsas periodontales fueron Fusobacterium (50 %), Prevotella intermedia/nigrescens (40 %), Campilobacter rectus (40 %), Eikenella corrodens (36,7 %), bacilos entéricos Gram negativos (36,7 %), Porphyromonas gingivalis (33,3 %) y Eubacterium (33,3 %). Los agentes patógenos aislados de la válvula aórtica fueron Propionibacterium acnes (12 %), bacilos entéricos Gram negativos (8 %), Bacteroides merdae (4 %) y Clostridium bifermentans (4 %), y de la válvula mitral, P. acnes y Clostridium beijerinckii. La PCR convencional no arrojó resultados positivos para agentes patógenos orales y solo se detectó ADN bacteriano en dos muestras. Conclusiones. La microbiota periodontal de pacientes sometidos a cirugía de reemplazo valvular estaba conformada por especies Gram negativas que han sido relacionadas con infecciones en tejidos extraorales; sin embargo, no se encontraron agentes patógenos periodontales en los tejidos de las válvulas. Aunque hubo muestras de estos tejidos y subgingivales, positivas para bacilos entéricos Gram negativos, no es posible asegurar que tuvieran el mismo origen filogenético.

Rift Valley fever virus (RVFV) is a mosquito-borne bunyavirus that causes an important disease in ruminants, with great economic losses. The infection can be also transmitted to humans; therefore, it is considered a major threat to both human and animal health. In a previous work, we described a novel RVFV variant selected in cell culture in the presence of the antiviral agent favipiravir that was highly attenuated in vivo. This variant displayed 24 amino acid substitutions in different viral proteins when compared to its parental viral strain, two of them located in the NSs protein that is known to be the major virulence factor of RVFV. By means of a reverse genetics system, in this work we have analyzed the effect that one of these substitutions, P82L, has in viral attenuation in vivo. Rescued viruses carrying this single amino acid change were clearly attenuated in BALB/c mice while their growth in an interferon (IFN)-competent cell line as well as the production of interferon beta (IFN-β) did not seem to be affected. However, the pattern of nuclear NSs accumulation was modified in cells infected with the mutant viruses. These results highlight the key role of the NSs protein in the modulation of viral infectivity.

Riboflavin transporter deficiency (RTD) is a childhood-onset neurodegenerative disorder characterized by sensorineural deafness and motor neuron degeneration. Since riboflavin plays key functions in biological oxidation-reduction reactions, energy metabolism pathways involving flavoproteins are affected in RTD. We recently generated induced pluripotent stem cell (iPSC) lines from affected individuals as an in vitro model of the disease and documented mitochondrial impairment in these cells, dramatically impacting cell redox status. This work extends our study to motor neurons (MNs), i.e., the cell type most affected in patients with RTD. Altered intracellular distribution of mitochondria was detected by confocal microscopic analysis (following immunofluorescence for superoxide dismutase 2 (SOD2), as a dual mitochondrial and antioxidant marker), and βIII-Tubulin, as a neuronal marker. We demonstrate significantly lower SOD2 levels in RTD MNs, as compared to their healthy counterparts. Mitochondrial ultrastructural abnormalities were also assessed by focused ion beam/scanning electron microscopy. Moreover, we investigated the effects of combination treatment using riboflavin and N-acetylcysteine, which is a widely employed antioxidant. Overall, our findings further support the potential of patient-specific RTD models and provide evidence of mitochondrial alterations in RTD-related iPSC-derived MNs—emphasizing oxidative stress involvement in this rare disease. We also provide new clues for possible therapeutic strategies aimed at correcting mitochondrial defects, based on the use of antioxidants.

Bluetongue virus (BTV), the prototype member of the genus Orbivirus (family Reoviridae), is the causative agent of an important livestock disease, bluetongue (BT), which is transmitted via biting midges of the genus Culicoides. To date, up to 29 serotypes of BTV have been described, which are classified as classical (BTV 1–24) or atypical (serotypes 25–27), and its distribution has been expanding since 1998, with important outbreaks in the Mediterranean Basin and devastating incursions in Northern and Western Europe. Classical vaccine approaches, such as live-attenuated and inactivated vaccines, have been used as prophylactic measures to control BT through the years. However, these vaccine approaches fail to address important matters like vaccine safety profile, effectiveness, induction of a cross-protective immune response among serotypes, and implementation of a DIVA (differentiation of infected from vaccinated animals) strategy. In this context, a wide range of recombinant vaccine prototypes against BTV, ranging from subunit vaccines to recombinant viral vector vaccines, have been investigated. This article offers a comprehensive outline of the live viral vectors used against BTV.

The underlying relation between Parkinson’s disease (PD) etiopathology and its major risk factor, aging, is largely unknown. In light of the causative link between genome stability and aging, we investigate a possible nexus between DNA damage accumulation, aging, and PD by assessing aging-related DNA repair pathways in laboratory animal models and humans. We demonstrate that dermal fibroblasts from PD patients display flawed nucleotide excision repair (NER) capacity and that Ercc1 mutant mice with mildly compromised NER exhibit typical PD-like pathological alterations, including decreased striatal dopaminergic innervation, increased phospho-synuclein levels, and defects in mitochondrial respiration. Ercc1 mouse mutants are also more sensitive to the prototypical PD toxin MPTP, and their transcriptomic landscape shares important similarities with that of PD patients. Our results demonstrate that specific defects in DNA repair impact the dopaminergic system and are associated with human PD pathology and might therefore constitute an age-related risk factor for PD.

Introducción: En el 2010, la Pontificia Universidad Javeriana Cali (PUJ Cali) ofertó por primera vez la carrera de Medicina. Debido a que el programa académico es relativamente nuevo y ofrece un plan de estudios innovador, basado en un currículo por competencias que integra las ciencias básicas, las clínicas médicas y la salud pública, es necesario verificar que el perfil de ingreso propuesto de los neojaverianos se adapte a las exigencias académicas, científicas y humanas requerida en la formación de los médicos javerianos. Objetivo: Caracterizar a los estudiantes del programa de medicina de la PUJ Cali, a partir de aspectos sociodemográficos. Materiales y métodos: Estudio descriptivo de corte transversal que caracterizó a los estudiantes de Medicina de la PUJ Cali. De los 423 estudiantes matriculados en el primer periodo (enero-junio) de 2013, 299 diligenciaron una encuesta diseñada en el software Cardiff TELEform® versión 10.0. Resultados: De los 299 estudiantes que diligenciaron la encuesta, el 64,3 % son mujeres y el 35,7 % son hombres; el 90,9 % son mayores de 18 años; el 59 % nacieron en la ciudad de Cali. La vivienda se distribuye en el 69,9 % en los estratos 4, 5 y 6. Conclusión: La muestra estudiada corresponde a una población homogénea donde un poco más de la mitad de los estudiantes nacieron y residen en la ciudad de Cali. Asimismo, los estratos socioeconómicos predominantes son el 4, 5 y 6, teniendo en cuen- ta que una cantidad considerable de estudiantes desconocen el estrato donde residen.

Introducción: La técnica anatómica de diafanización consiste en transparentar los tejidos blandos de organismos vertebrados (aclaramiento), para teñir los tejidos mineralizados y poder visualizar los componentes óseos y cartilaginosos (tinción). Objetivo: Revisar los reportes disponibles en la literatura especializada que han descrito el desarrollo embrionario del sistema óseo de mamíferos a través de técnicas de diafanización y tinción simple o tinción doble. Materiales y métodos: Se revisó la literatura sobre el tema en PubMed, Google Académico y SciELO, basándose en la metodología PRISMA, a través de la asociación del término double staining con los descriptores en salud del Medical Subject Headings (MeSH) alizarin red, alcian blue, bone y cartilage , y la combinación con los operadores boleanos + y &. Resultados: E n esta revisión de la literatura se incluyeron 22 artículos que describieron la técnica de diafanización y tinción simple o tinción doble empleada en la observación, registro y análisis de desarrollo embrionario del sistema óseo de ma - míferos. Conclusión: La diafanización y tinción simple o tinción doble es una técnica anatómica de estudio —versátil y de bajo costo— del de - sarrollo embrionario del sistema óseo, la cual puede emplearse en estudios toxicológicos, para descartar la posibilidad de anomalías de desarro- llo durante la formación del cartílago (condrogé - nesis) y del hueso (osteogénesis), asociadas a la exposición de un posible agente teratógeno.

Introduction: protostylid is a non-metric dental trait observable on the vestibular surface of first and second deciduous and permanent molars in populations of Mongoloid origin. The objective of this literature review was to identify the expression of protostylid in human populations from Southwestern Colombia to help clarify the macro-evolutionary processes in this geographic region. Methods: a systematic literature review in PubMed, Google Scholar, and SciELO using the following medical descriptors: “dental morphology”, “non-metric dental traits”, and “protostylid”, combined with the Boolean operators “+” and “&” by the PRISMA methodology, in order to find publications describing the prevalence and variability of protostylid in human populations from Southwestern Colombia by means of the Arizona State University Dental Anthropology System. Results: 11 publications were found allowing us to obtain biological matrix distances based on cluster hierarchical classification and their respective dendrograms using the Ward method, which allowed us to observe most of the populations in Southwestern Colombia considered in this study and their relationships with other world populations. Conclusions: miscegenation processes have influenced the prevalence of protostylid, decreasing the expression of groove, blunt vertex cusp, and free vertex cusp in indigenous populations, and increasing the expression of fossa or P point in mixed coastal populations and populations of African descent. It also became clear that the expression of fossa or P point can occur in the absence or presence of protostylid.

Mitochondrial dysfunction is a key element in the pathogenesis of neurodegenerative disorders, such as riboflavin transporter deficiency (RTD). This is a rare, childhood-onset disease characterized by motoneuron degeneration and caused by mutations in SLC52A2 and SLC52A3, encoding riboflavin (RF) transporters (RFVT2 and RFVT3, respectively), resulting in muscle weakness, ponto-bulbar paralysis and sensorineural deafness. Based on previous findings, which document the contribution of oxidative stress in RTD pathogenesis, we tested possible beneficial effects of several antioxidants (Vitamin C, Idebenone, Coenzyme Q10 and EPI-743, either alone or in combination with RF) on the morphology and function of neurons derived from induced pluripotent stem cells (iPSCs) from two RTD patients. To identify possible improvement of the neuronal morphotype, neurite length was measured by confocal microscopy after β-III tubulin immunofluorescent staining. Neuronal function was evaluated by determining superoxide anion generation by MitoSOX assay and intracellular calcium (Ca2+) levels, using the Fluo-4 probe. Among the antioxidants tested, EPI-743 restored the redox status, improved neurite length and ameliorated intracellular calcium influx into RTD motoneurons. In conclusion, we suggest that antioxidant supplementation may have a role in RTD treatment.

The sequence of non-structural protein NS1 of bluetongue virus (BTV), which contains immunodominant CD8+ T cell epitopes, is highly conserved among BTV serotypes, and has therefore become a major tool in the development of a universal BTV vaccine. In this work, we have engineered multiserotype BTV vaccine candidates based on recombinant chimpanzee adenovirus (ChAdOx1) and modified vaccinia virus Ankara (MVA) vectors expressing the NS1 protein of BTV-4 or its truncated form NS1-Nt. A single dose of ChAdOx1-NS1 or ChAdOx1-NS1-Nt induced a moderate CD8+ T cell response and protected IFNAR(-/-) mice against a lethal dose of BTV-4/MOR09, a reassortant strain between BTV-1 and BTV-4, although the animals showed low viremia after infection. Furthermore, IFNAR(-/-) mice immunized with a single dose of ChAdOx1-NS1 were protected after challenge with a lethal dose of BTV-8 in absence of viremia nor clinical signs. Additionally, the heterologous prime-boost ChAdOx1/MVA expressing NS1 or NS1-Nt elicited a robust NS1 specific CD8+ T cell response and protected the animals against BTV-4/MOR09 even 16 weeks after immunization, with undetectable levels of viremia at any time after challenge. Subsequently, the best immunization strategy based on ChAdOx1/MVA-NS1 was assayed in sheep. Non-immunized animals presented fever and viremia levels up to 104 PFU/mL after infection. In contrast, although viremia was detected in immunized sheep, the level of virus in blood was 100 times lower than in non-immunized animals in absence of clinical signs.

In vitro neutralizing antibodies have been often correlated with protection against Rift Valley fever virus (RVFV) infection. We have reported previously that a single inoculation of sucrose-purified modified vaccinia Ankara (MVA) encoding RVFV glycoproteins (rMVAGnGc) was sufficient to induce a protective immune response in mice after a lethal RVFV challenge. Protection was related to the presence of glycoprotein specific CD8+ cells, with a low-level detection of in vitro neutralizing antibodies. In this work we extended those observations aimed to explore the role of humoral responses after MVA vaccination and to study the contribution of each glycoprotein antigen to the protective efficacy. Thus, we tested the efficacy and immune responses in BALB/c mice of recombinant MVA viruses expressing either glycoprotein Gn (rMVAGn) or Gc (rMVAGc). In the absence of serum neutralizing antibodies, our data strongly suggest that protection of vaccinated mice upon the RVFV challenge can be achieved by the activation of cellular responses mainly directed against Gc epitopes. The involvement of cellular immunity was stressed by the fact that protection of mice was strain dependent. Furthermore, our data suggest that the rMVA based single dose vaccination elicits suboptimal humoral immune responses against Gn antigen since disease in mice was exacerbated upon virus challenge in the presence of rMVAGnGc or rMVAGn immune serum. Thus, Gc-specific cellular immunity could be an important component in the protection after the challenge observed in BALB/c mice, contributing to the elimination of infected cells reducing morbidity and mortality and counteracting the deleterious effect of a subneutralizing antibody immune response.

La periodontitis es una enfermedad infecciosa e inflamatoria crónica, multifactorial y compleja que se ha relacionado con diferentes complicaciones sistémicas, entre ellas la enfermedad cardiovascular. Esta asociación ha sido difícil de comprobar en estudios epidemiológicos, ya que presentan sesgos o factores de riesgo clásicos además, la enfermedad cardiovascular comprende una diversidad de patologías también de origen multifactorial, lo que hace aún más complejo determinar su relación causaefecto. Los estudios reportados en la literatura que intentan explicar la relación entre la enfermedad periodontal y la enfermedad cardiovascular, desde sus diferentes diseños metodológicos, muestran suficiente evidencia científica para considerar la enfermedad periodontal como un indicador de riesgo cardiovascular, información supremamente necesaria para la comunidad médica y odontológica, quienes en su práctica clínica deben incluir protocolos de trabajo interdisciplinario, a fin de procurar un manejo integral del individuo.

Arboviruses are arthropod-borne viruses that exhibit worldwide distribution and are a constant threat, not only for public health but also for wildlife, domestic animals, and even plants. To study disease pathogenesis and to develop efficient and safe therapies, the use of an appropriate animal model is a critical concern. Adult mice with gene knockouts of the interferon α/β (IFN-α/β) receptor (IFNAR(−/−)) have been described as a model of arbovirus infections. Studies with the natural hosts of these viruses are limited by financial and ethical issues, and in some cases, the need to have facilities with a biosafety level 3 with sufficient space to accommodate large animals. Moreover, the number of animals in the experiments must provide results with statistical significance. Recent advances in animal models in the last decade among other gaps in knowledge have contributed to the better understanding of arbovirus infections. A tremendous advantage of the IFNAR(−/−) mouse model is the availability of a wide variety of reagents that can be used to study many aspects of the immune response to the virus. Although extrapolation of findings in mice to natural hosts must be done with care due to differences in the biology between mouse and humans, experimental infections of IFNAR(−/−) mice with several studied arboviruses closely mimics hallmarks of these viruses in their natural host. Therefore, IFNAR(−/−) mice are a good model to facilitate studies on arbovirus transmission, pathogenesis, virulence, and the protective efficacy of new vaccines. In this review article, the most important arboviruses that have been studied using the IFNAR(−/−) mouse model will be reviewed.

The perception of colour and its subjective effects are key issues to designing safe and enjoyable bike lanes. This paper addresses the relationship between the colours of bike lane interventions-in particular pavement painting and intersection design-and the subjective evaluation of liking, visual saliency, and perceived safety related to such an intervention. Utilising images of three real bike lane intersections modified by software to change their colour (five in total), this study recruited 538 participants to assess their perception of all fifteen colour-design combinations. A multivariate analysis of covariance (MANCOVA) with the Bonferroni post hoc test was performed to assess the effect of the main conditions (colour and design) on the dependent variables (liking towards the intervention, level of visual saliency of the intersection, and perceived safety of the bike lane). The results showed that the colour red was more positively associated to the outcome variables, followed by yellow and blue. Additionally, it was observed that the effect of colour widely outweighs the effect of design, suggesting that the right choice and use of colour would increase the effectiveness on bike-lanes pavement interventions. Limitations and future directions are discussed.

The endoplasmic reticulum (ER) stress-mediated pathway is involved in a wide range of human neurodegenerative disorders. Hence, molecules that regulate the ER stress response represent potential candidates as drug targets to tackle these diseases. In previous studies we demonstrated that upon acetylation the reticulon-1C (RTN-1C) variant of the reticulon family leads to inhibition of histone deacetylase (HDAC) enzymatic activity and endoplasmic reticulum stress-dependent apoptosis. Here, by microarray analysis of the whole human genome we found that RTN-1C is able to specifically regulate gene expression, modulating transcript clusters which have been implicated in the onset of neurodegenerative disorders. Interestingly, we show that some of the identified genes were also modulated in vivo in a brain-specific mouse model overxpressing RTN-1C. These data provide a basis for further investigation of RTN-1C as a potential molecular target for use in therapy and as a specific marker for neurological diseases.

Introducción. Quibdó, capital del departamento del Chocó, se ha caracterizado por ser una de las regiones más deprimidas en Colombia. La presencia de virus entéricos tipo rotavirus y virus de la hepatitis A en el agua de consumo, además de ser un indicador de su mala calidad, es una importante fuente de contaminación para los individuos que la consumen. Objetivo. Demostrar la presencia de estos dos agentes virales en el agua de consumo para contribuir con la explicación de la morbilidad por enfermedad diarreica aguda y hepatitis en la región, y aclarar que el origen de estos virus es por contaminación de desechos humanos más que por la materia fecal de bovinos o porcinos. Materiales y métodos. Se procesaron 4.000 litros de agua que se llevaron a ultrafiltración tangencial y a centrifugación con filtros Centriprep Ultracel YM-50. Se aplicó la prueba de RT-PCR a 60 muestras de agua no tratada y a 20 muestras de agua tratada por el acueducto. Las muestras positivas fueron secuenciadas y con el análisis de dichas secuencias se elaboraron árboles filogenéticos. Resultados. Seis de las muestras resultaron positivas para rotavirus y dos más para virus de la hepatitis A. Éstos aparecieron tanto en aguas tratadas como no tratadas. Los análisis filogenéticos demostraron que el rotavirus pertenece al serotipo G2 humano y que el virus de la hepatitis A fue también de origen humano. Conclusión. El agua analizada presenta un alto nivel de contaminación, demostrado por la presencia de virus patógenos para el hombre.

Recent studies indicate that molecular signals from microglia determine disease progression in transgenic mice overexpressing mutant superoxide dismutase (mutSOD1) typical of amyotrophic lateral sclerosis patients and that toxicity of mutSOD1 in motor neurons descends from its tendency to associate with mitochondria.To assess whether the neurotoxicity of mutSOD1 is influenced by signals from glia, we challenged motoneuronal cells overexpressing either wild-type or mutant SOD1 with inflammatory cytokines. We have obtained evidence that combined treatment with tumor necrosis factor α and interferon γ increases the fraction of both wtSOD1 and mutSOD1 associated with mitochondria, but these inflammatory cytokines dramatically alter morphological features and functionality of mitochondria only in cells expressing mutSOD1. As an effect downstream the increase in mitochondria-associated mutSOD1, the ratio between reduced and oxidized glutathione further shifts toward the oxidized form in this compartment and a clear death phenotype is evoked upon treatment with inflammatory cytokines. These results suggest that signals coming from non-neuronal cells contribute to death of motor neurons induced by mutSOD1 through reinforcement of mitochondrial damage.