Engineered PROTAC-CID Systems for Mammalian Inducible Gene Regulation.

Gene regulation via chemically induced dimerization (CID) is useful for biomedical research. However, the number, type, versatility, and in vivo applications of CID tools remain limited. Here, we demonstrate the development of proteolysis-targeting chimera-based scalable CID (PROTAC-CID) platforms by systematically engineering the available PROTAC systems for inducible gene regulation and gene editing. Further, we show orthogonal PROTAC-CIDs that can fine-tune gene expression at gradient levels or multiplex biological signals with different logic gating operations. Coupling the PROTAC-CID platform with genetic circuits, we achieve digitally inducible expression of DNA recombinases, base- and prime-editors for transient genome manipulation. Finally, we package a compact PROTAC-CID system into adeno-associated viral vectors for inducible and reversible gene activation in vivo. This work provides a versatile molecular toolbox that expands the scope of chemically inducible gene regulation in human cells and mice.

Coincidencia de Rhupus y embarazo. Reporte de caso / Coincidence of Rhupus and pregnancy. Case report

Resumen ANTECEDENTES: Si bien el lupus eritematoso sistémico y la artritis reumatoide son enfermedades suficientemente descritas, no lo es la asociación de estas, que se denomina rhupus, que hace referencia a la manifestación clínica resultante de ambas enfermedades enmascaradas que dificulta el diagnóstico y tratamiento. CASO CLÍNICO: Paciente de 37 años que inició con un cuadro con características de lupus eritematoso sistémico y artritis reumatoide hacía 16 años. Se encontró con alteraciones en los estudios de laboratorio y con hallazgos radiológicos que apoyaron el cuadro de rhupus. Por lo anterior se documenta la evolución de este padecimiento, que coincide con su embarazo y después de éste. CONCLUSIONES: La aparición simultánea de lupus eritematoso sistémico y artritis reumatoide, aun cuando fue reportada desde hace décadas, es una enfermedad rara en frecuencia, por lo que hay escasa información del rhupus solo y más aún en coincidencia con el embarazo.

Abstract BACKGROUND: Although systemic lupus erythematosus and rheumatoid arthritis are sufficiently described diseases, the association of these is not, and is called rhupus, which refers to the clinical manifestation resulting from both diseases masked, making diagnosis and treatment difficult. CLINICAL CASE: 37-year-old patient who started with a clinical picture with features of systemic lupus erythematosus and rheumatoid arthritis 16 years ago. She was found to have alterations in laboratory studies and radiological findings that supported the diagnosis of rheupus. Therefore, the evolution of this condition is documented, which coincides with and after her pregnancy. CONCLUSIONS: The simultaneous occurrence of systemic lupus erythematosus and rheumatoid arthritis, even though it was reported decades ago, is a rare disease in frequency, so there is scarce information on rhupus alone and even more so in coincidence with pregnancy.

Differential activation of innate and adaptive lymphocytes during latent or active infection with Mycobacterium tuberculosis.

Most individuals infected with Mycobacterium tuberculosis (Mtb) have latent tuberculosis (TB), which can be diagnosed with tests (such as the QuantiFERON-TB Gold test [QFT]) that detect the production of IFN-γ by memory T cells in response to the Mtb-specific antigens 6 kDa early secretory antigenic target EsxA (Rv3875) (ESAT-6), 10 kDa culture filtrate antigen EsxB (Rv3874) (CFP-10), and Mtb antigen of 7.7 kDa (Rv2654c) (TB7.7). However, the immunological mechanisms that determine if an individual will develop latent or active TB remain incompletely understood. Here we compared the response of innate and adaptive peripheral blood lymphocytes from healthy individuals without Mtb infection (QFT negative) and from individuals with latent (QFT positive) or active TB infection, to determine the characteristics of these cells that correlate with each condition. In active TB patients, the levels of IFN-γ that were produced in response to Mtb-specific antigens had high positive correlations with IL-1ß, TNF-α, MCP-1, IL-6, IL-12p70, and IL-23, while the proinflammatory cytokines had high positive correlations between themselves and with IL-12p70 and IL-23. These correlations were not observed in QFT-negative or QFT-positive healthy volunteers. Activation with Mtb-soluble extract (a mixture of Mtb antigens and pathogen-associated molecular patterns [PAMPs]) increased the percentage of IFN-γ-/IL-17-producing NK cells and of IL-17-producing innate lymphoid cell 3 (ILC3) in the peripheral blood of active TB patients, but not of QFT-negative or QFT-positive healthy volunteers. Thus, active TB patients have both adaptive and innate lymphocyte subsets that produce characteristic cytokine profiles in response to Mtb-specific antigens or PAMPs. These profiles are not observed in uninfected individuals or in individuals with latent TB, suggesting that they are a response to active TB infection.

A monocyte/dendritic cell molecular signature of SARS-CoV-2-related multisystem inflammatory syndrome in children with severe myocarditis.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children is generally milder than in adults, but a proportion of cases result in hyperinflammatory conditions often including myocarditis. METHODS: To better understand these cases, we applied a multiparametric approach to the study of blood cells of 56 children hospitalized with suspicion of SARS-CoV-2 infection. Plasma cytokine and chemokine levels and blood cellular composition were measured, alongside gene expression at the bulk and single-cell levels. FINDINGS: The most severe forms of multisystem inflammatory syndrome in children (MIS-C) related to SARS-CoV-2 that resulted in myocarditis were characterized by elevated levels of pro-angiogenesis cytokines and several chemokines. Single-cell transcriptomics analyses identified a unique monocyte/dendritic cell gene signature that correlated with the occurrence of severe myocarditis characterized by sustained nuclear factor κB (NF-κB) activity and tumor necrosis factor alpha (TNF-α) signaling and associated with decreased gene expression of NF-κB inhibitors. We also found a weak response to type I and type II interferons, hyperinflammation, and response to oxidative stress related to increased HIF-1α and Vascular endothelial growth factor (VEGF) signaling. CONCLUSIONS: These results provide potential for a better understanding of disease pathophysiology. FUNDING: Agence National de la Recherche (Institut Hospitalo-Universitaire Imagine, grant ANR-10-IAHU-01; Recherche Hospitalo-Universitaire, grant ANR-18-RHUS-0010; Laboratoire d'Excellence ''Milieu Intérieur," grant ANR-10-LABX-69-01; ANR-flash Covid19 "AIROCovid" and "CoVarImm"), Institut National de la Santé et de la Recherche Médicale (INSERM), and the "URGENCE COVID-19" fundraising campaign of Institut Pasteur.

Obesidad y características de personalidad en adolescentes de Argentina y España: Un estudio transcultural / Obesity and personality characteristics in adolescents from Argentina and Spain: A cross-cultural study

Resumen Los objetivos de este estudio fueron: 1. Comparar la presencia de sobrepeso y obesidad en adolescentes de Argentina vs. España, 2. Comparar la personalidad según diagnóstico nutricional (DN) y lugar de residencia (LR), y 3. Identificar la presencia de riesgo psicopatológico (RP). Participaron 272 adolescentes, hombres y mujeres, de 11-16 años de edad, quienes completaron el Cuestionario de Personalidad para Niños de Eysenck (EPQ-J) y, para calcular el índice de masa corporal, fueron recabadas medidas de talla y peso. Presentaron sobrepeso u obesidad 39% de los adolescentes españoles y 43% de los argentinos, con quienes se conformaron dos grupos clínicos, uno por país; mientras que ambos grupos control estuvieron constituidos por los participantes normopeso. La combinación de las variables DN y LR no arrojó diferencias significativas en cuanto a las dimensiones de personalidad. Sin embargo, ambos grupos clínicos presentaron mayor neuroticismo (NE) que sus pares control. Respecto al LR, los adolescentes argentinos registraron mayor NE y dureza emocional (4% de alto riesgo) vs.3% de alto riesgo por NE en los españoles. Los datos obtenidos no permitieron establecer un perfil psicopatológico del adolescente con sobrepeso u obesidad, dado que prevaleció la influencia del país de procedencia.

Abstract The objectives of this study were: 1. To compare the presence of overweight and obesity in adolescents from Argentina vs. Spain, 2. To compare personality according to nutritional diagnosis (ND) and place of residence (PR), and 3. To identify the presence of psychopathological risk (PR). A total of 272 adolescents, men and women from 11 to 16 years completed the Personality Questionnaire for Children of Eysenck (EPQ-J) and body mass index was calculated through height and weight measures. The 39% of the Spanish adolescents and 43% of the Argentinians presented overweight or obesity, two clinical groups were formed, one per country and both control groups were constituted by the normal weight participants. The combination of the variables ND and PR did not show significant differences in terms of personality dimensions. However, both clinical groups presented greater neuroticism (NE) than their control peers. Regarding the PR, the Argentine adolescents registered higher NE and emotional hardness (4% of high risk) than 3% of high risk of NE in the Spanish. The data obtained did not allow to establish a psychopathological profile of the overweight or obese adolescent, this is because the influence of the country of origin prevailed.

Implant treatment in immunosuppressed renal transplant patients: A prospective case-controlled study.

OBJECTIVES: The aim of this clinical study was to evaluate long-term outcomes of implant therapy in a cohort of immunosuppressed renal transplant patients compared with a matched control group. MATERIAL AND METHODS: Pharmacologically immunosuppressed renal transplant patients received dental implant treatment between 2001 and 2011. Periodontal, clinical and radiographic parameters were prospectively measured with a mean follow-up of 116.8 months (range from 84 to 192 months). A matched controlled non-transplant sample receiving similar implant treatment in the same time was included as a control group. RESULTS: Implant survival rate was over 98% in both test and control groups (100% and 98.84%, respectively). Peri-implant mucositis was diagnosed in 46.80% of the implants in the study group and in 48.80% in the control group. Peri-implantitis occurred in 5.10% of the implants in the study group and in 8.10% of the controls. Wound healing and post-operative pain were similar in both groups. CONCLUSIONS: Despite the limitations of this study, pharmacological immunosuppression in renal transplant patients did not affect implant outcomes. Renal transplant patients should be carefully controlled periodically after implant treatment. CLINICAL IMPLICATIONS: The results from this investigation justify the use of dental implants for the dentalrehabilitation of immunosuppressed patients after renal transplantation provided they follow the necessarylong-term monitoring and regular maintenance of their oral and systemic health.

Extracellular Vesicles Released from Mycobacterium tuberculosis-Infected Neutrophils Promote Macrophage Autophagy and Decrease Intracellular Mycobacterial Survival.

Tuberculosis is an infectious disease caused by Mycobacterium tuberculosis (Mtb). In the lungs, macrophages and neutrophils are the first immune cells that have contact with the infecting mycobacteria. Neutrophils are phagocytic cells that kill microorganisms through several mechanisms, which include the lytic enzymes and antimicrobial peptides that are found in their lysosomes, and the production of reactive oxygen species. Neutrophils also release extracellular vesicles (EVs) (100-1,000 nm in diameter) to the extracellular milieu; these EVs consist of a lipid bilayer surrounding a hydrophilic core and participate in intercellular communication. We previously demonstrated that human neutrophils infected in vitro with Mtb H37Rv release EVs (EV-TB), but the effect of these EVs on other cells relevant for the control of Mtb infection, such as macrophages, has not been completely analyzed. In this study, we characterized the EVs produced by non-stimulated human neutrophils (EV-NS), and the EVs produced by neutrophils stimulated with an activator (PMA), a peptide derived from bacterial proteins (fMLF) or Mtb, and observed that the four EVs differed in their size. Ligands for toll-like receptor (TLR) 2/6 were detected in EV-TB, and these EVs favored a modest increase in the expression of the co-stimulatory molecules CD80, a higher expression of CD86, and the production of higher amounts of TNF-α and IL-6, and of lower amounts of TGF-ß, in autologous human macrophages, compared with the other EVs. EV-TB reduced the amount of intracellular Mtb in macrophages, and increased superoxide anion production in these cells. TLR2/6 ligation and superoxide anion production are known inducers of autophagy; accordingly, we found that EV-TB induced higher expression of the autophagy-related marker LC3-II in macrophages, and the co-localization of LC3-II with Mtb inside infected macrophages. The intracellular mycobacterial load increased when autophagy was inhibited with wortmannin in these cells. In conclusion, our results demonstrate that neutrophils produce different EVs in response to diverse activators, and that EV-TB activate macrophages and promote the clearance of intracellular Mtb through early superoxide anion production and autophagy induction, which is a novel role for neutrophil-derived EVs in the immune response to Mtb.

Implant treatment in pharmacologically immunosuppressed liver transplant patients: A prospective-controlled study.

OBJECTIVES: The main objective of this prospective study was to evaluate the long-term outcome of implant therapy in liver transplant patients (LTP). The secondary goal was to assess several implant- and patient-dependent variables, such as peri-implantitis (PI), peri-implant mucositis (PIM), bone loss (BL), and immediate postoperative complications. MATERIAL AND METHODS: Two groups, including 16 pharmacologically immunosuppressed LTP and 16 matched controls, received 52 and 54 implants, respectively, between 1999 and 2008. After evaluating the postoperative healing, a mean follow-up of more than 8 years was carried out, and radiographic, clinical, and periodontal parameters were recorded to evaluate implant survival and implant- and patient-dependent outcomes. RESULTS: The early postsurgical complications were similar in both groups. Implant survival rate was 100% in the LTP group and 98.15% in the CG. PIM was diagnosed in 35.42% of the implants and 64.29% of the patients of LTP group (LTPG) and in 43.40% of the implants and 56.25% of the patients in the CG. PI was detected in 4.17% of the implants and 7.10% of the patients in the LTPG and in 9.43% of the implants and 18.80% of the patients in the CG. CONCLUSION: Pharmacologically immunosuppression in liver transplant patients was not a risk factor for implant failure, nor for the incidence of peri-implant diseases. Liver transplant is not a contraindication for dental implant treatment, although these patients should be carefully monitored during follow-up care.

Surgical Treatment of a Peripheral Ossifying Fibroma and Reconstruction with a Porcine Collagen Matrix: A Case Report.

A 35-year-old woman was referred to the Department of Oral Medicine and Orofacial Surgery after several recurrences of an ossifying fibroma (OF) that affected the free and attached gingiva of the maxillary right central incisor. Surgery was performed with a complete excision of the lesion together with the surrounding healthy tissue up to the bone. To guide the healing of the anterior esthetic framework and the excised tissues, a porcine collagen matrix as an alternative to connective tissue graft was used. After an 18-month follow-up, the lesion had not recurred and keratinized gingiva had formed around the area.

Determinación de los niveles séricos de vitamina D en los pacientes diagnosticados con Esclerosis múltiple y su relación con la severidad y el comportamiento de la enfermedad en el "Hospital Metropolitano de Quito" y en el "Hospital Carlos Andrade Marín" / Determination of vitamin D serum levels in patients diagnosed with multiple sclerosis and their relationship with the severity and illness behavior in the Metropolitan Hospital in Quito and Carlos Andrade Marín Hospital

Introducción: El estudio pretende relacionar los niveles séricos de vitamina D, en pacientes con Esclerosis Múltiple (EM), con la severidad y el comportamiento de la enfermedad. Materiales y Métodos: Reclutamos 81 pacientes con EM, de los cuales se obtuvo una muestra de 50 pacientes para compararlos con 50 sujetos sanos, pareados por edad, sexo y color de la piel. La severidad de la enfermedad se evaluó con la escala de Kurtzke, número de recaídas, exposición solar, etc. Resultados: La concentración sérica de vitamina D en pacientes con EM (M=31.9 SD=12.3 ng/ml) fue similar a la de los controles (M=30.3 SD= 8.0 ng/ml; p=0.53). La prevalencia de insuficiencia de vitamina D fue del 42% (n=21) entre los casos y 46% (n=23) entre los controles, p>0.05). Se calculó la relación entre los niveles óptimos de Vitamina D (> 40, ng/ml) con un bajo grado de discapacidad, definida como un valor en la escala de Kurtzke < 3.5 [χ2(1, N=100)= 3.13 p=0.3]. El promedio de vitamina D en los pacientes con más de una recaída fue de 31.0 ng /ml y con menos de una recaída, 32.5 ng/ml fue similar (p=0.66). Discusión: No hubo diferencia significativa en las concentraciones séricas de vitamina D en pacientes con EM y sujetos sanos, pareados por género, edad y color de la piel. No hubo correlación entre el nivel de vitamina D y el grado de discapacidad medido por la escala de Kurtzke ni con el número de recaídas.

Introduction: The study seeks to relate vitamin D serum levels in Multiple Sclerosis (MS) patients with disease severity and its progression. Methods: From eighty one MS patients enrolled, fifty were selected to compare to healthy subjects, paired according to age, gender and skin color. Disease severity was assessed using the Kurtzke severity scale, relapses number, solar exposure. Results: The mean vitamin D serum concentration in MS patients (M= 31.9 SD=12.3 ng/ml) was similar to controls' (M=30.31 SD=8.0 ng/ml; p=0.53). The prevalence of vitamin D insufficiency was 42% (n=22) among cases and 46% (n=23) among controls (p=0.91). The relationship between an optimal vitamin D serum concentration (>40 ng/ml) and a low grade of disability, using a cutoff point in the Kurtzke scale < 3.5, was assessed and a non-significant correlation was found [χ2(1, N=100)= 3.13 p=0.3]. The mean vitamin D level among patients with more than one relapse (M=31.0 ng/ml) was similar to those with less than one relapse (M=32.49, p=0.66). Discusion: There was no difference of vitamin D serum levels between MS patients and the general population. A weak and not significant correlation was found with the degree of disability, measured by the Kurtzke severity scale, and also with the number of relapses.

Successful treatment of oral pemphigus vulgaris in an insulin-dependant geriatric patient.

OBJECTIVE: To present a clinical report of a patient treated with corticosteroids for oral pemphigus vulgaris (PV) lesions. BACKGROUND: PV is the type of pemphigus that most often affects the oral mucosa and tends not to appear in elderly people. METHODS: Two biopsies were needed for diagnosis. She was treated with oral prednisone and topically with 0.05% clobetasol propionate. CONCLUSION: An early diagnosis and treatment is needed for a good prognosis, especially in elderly patients with multiple systemic pathology.

Efectividad del uso de alcohol glicerinado para la descontaminación de manos en una población sin acceso al agua potable posterremoto en Pisco, Perú / Effectiveness of glycerinated alcohol use for decontaminating hands in a population without access to safe water post earthquake in Pisco, Perú

Se evaluó la efectividad del uso de alcohol glicerinado en la desinfección de manos, de personas que preparaban los alimentos en ollas comunes para damnificados de albergues y comités vecinales sin acceso al agua potable, en los primeros días posteriores al terremoto de Pisco. Se realizó un preexperimento con 20 mujeres adultas que no se habían lavado las manos, se cuantificó la carga bacterianade aerobios mesófilos, enterobacterias y Staphylococcus aureus en sus manos antes y después de la aplicación de alcohol glicerinado, usando la prueba de Wilcoxon se encontró una reducción significativa (p menor que 0,001) de la carga bacteriana. En conclusión, la aplicación de alcohol glicerinado es efectiva para la desinfección de manos en manipuladores de alimentos en una población sin acceso al agua yposterremoto.

We assessed the effectiveness of the use of glycerinated alcohol for disinfecting the hands of persons fixing foods in communal kitchens in refugee and neighborhood camps with no access to potable water during the first days alter the earthquake that hit Pisco, Peru in August 2007. A previous assessment was performed in 20 adult women with no prior handwashing. Bacterial load in their hands, including mesophilic microorganisms, Enterobacteriaceae, and Staphylococcus aureus was measured before and after the use of glycerinated alcohol. A significant reduction in bacterial load (p minor that 0.001) was found according to the Wilcoxon test. We conclude that the use of glycerinated alcohol is effective for hand disinfection in people manipulating food in an area with no access to potable water alter a naturaldisaster, in this case, the earthquake that hit Pisco in August 2007.

Consideraciones bioéticas y código de salud para el Perú / Considerations bioethics and health code for Peru

El documento detalla información de la introducción; contexto y problemática de tránsito de la ética tradicional a la ética moderna; de algunas consideraciones a tener en cuenta para una bioética en el Perú de fines del Siglo XX e inicios del Siglo XXI; de la propuesta de código de salud del CMP, documento de trabajo para construir consensos y convencionalidades para un nuevo código de salud en el Perú

Políticas y estrategias del sector salud / Health sector politics and strategies

Define las políticas (salud integral, salud responsabilidad de todos, descentralización y regionalización, priorización del ámbito local, revalorización de los recursos humanos, desarrollo científico y producción tecnológica, medicamentos como bien social, recuperación de los establecimientos de salud); y las estrategias de salud (difusión y movilización social por la salud, concertación social para el cambio en salud, fortalecimiento y legitimación del rol rector de Ministerio de Salud, profundización del proceso de descentralización en salud, promoción de las condiciones laborales y recalificación de los recursos humanos, reorientación de las acción sanitaria, difusión e intercambio del conocimiento y de las experiencias sanitariasl, reorientación del financiamiento para el logro de la equidad)