Favipiravir y/o umifenovir en infección por COVID-19 / Favipiravir and / or umifenovir in COVID-19 infection

CONTEXTO CLÍNICO: La Enfermedad por el Coronavirus 2019 (COVID­19, por su sigla en inglés Coronavirus Disease 2019) es una enfermedad respiratoria de humanos producida por un nuevo coronavirus identificado con la sigla SARS-CoV-2. El 11 de marzo de 2020 la Organización Mundial de la Salud (OMS) declaro la COVID-19 como una pandemia. Desde ese momento hasta este 01 de abril su circulación se ha reportado en 205 países reportándose más de 800.000 casos y la muerte 40.000 personas. El período de incubación de la infección por 2019­nCoV es de 2 a 14 días. La mayor parte de los contagios se producen persona a persona, siendo altamente transmisible. La clínica varía desde casos asintomáticos a cuadros febriles con tos y dificultad respiratoria, neumonía y distrés respiratorio. También puede acompañarse de alteraciones gastrointestinales. En los casos con mal pronóstico, el paciente presenta un importante deterioro respiratorio en 4-8 días. Las imágenes radiológicas muestran generalmente neumonía focal o generalizada semejante al síndrome de distress respiratorio agudo. La mayoría de los casos graves requieren ingreso hospitalario, siendo mayoritariamente casos primarios en pacientes de edad avanzada y con comorbilidades (diabetes, enfermedad crónica renal, hipertensión, enfermedad cardiaca y enfermedad pulmonar crónica). La tasa media de letalidad de los pacientes ingresados a UTI es cercana al 49%, siendo los valores más elevados en pacientes masculinos de más de 50 años con comorbilidades múltiples. Actualmente el tratamiento de la COVID­19 es sintomático y de sostén no existiendo hasta el momento tratamiento farmacológico específico curativo. Los tratamientos que se han propuestos son: inhibidores de la ARN polimerasa dependiente de ARN (remdesivir, favipiramir), inhibidores de la neuraminidasa (oseltamivir), inhibidores de la proteasa (lopinavir/ritonavir, desulfura, inhibidores de la enzima convertidora de angiotensina 2, inhibidores de quinasa (imatinib, baricitinib, ribavirin), inmunomoduladores (plasma de convaleciente, anticuerpos anti receptor IL-6 como tocilizumab y otros como interferón , glucocorticoides , umifenovir, cloroquina, etc.). De los tratamientos antivirales, el remdesivir reduciría el tiempo de mejoría en pacientes con COVID-19 y está recomendada por el Instituto Nacional de Estados Unidos y la agencia nacional española de medicamentos para pacientes con cuadros respiratorios graves. Dada la falta de vacunas o tratamientos específicos para el nuevo coronavirus SARS-CoV-2, se postula el uso de favipiramir o umifenovir para el tratamiento de pacientes COVID-19 positivos como drogas antivirales que se usan en otros cuadros virales respiratorios. TECNOLOGÍA: El umifenovir es una droga propuesta y aprobada en China y Rusia como droga sintética administrada por vía oral para infecciones virales. Tiene efectos inmunomoduladores para prevenir y curar la gripe influenza A y B, así como la hepatitis B y C. El favipiravir es un inhibidor de la ARN polimerasa dependiente de ARN contra la gripe y varios otros virus de ARN 7. En influenza, el efecto beneficioso se ha atribuido a una disminución de la carga viral pulmonar y los niveles de TNF-alfa en las vías respiratorias. OBJETIVO: El objetivo del presente informe es evaluar la evidencia disponible acerca de la eficacia, seguridad y aspectos relacionados a las políticas de cobertura del uso de favipiravir o umifenovir en infección por COVID-19. MÉTODOS: Se realizó una búsqueda en las principales bases de datos bibliográficas, en buscadores genéricos de internet, y financiadores de salud. Se priorizó la inclusión de revisiones sistemáticas (RS), ensayos clínicos controlados aleatorizados (ECAs), evaluaciones de tecnologías sanitarias (ETS), evaluaciones económicas, guías de práctica clínica (GPC) y recomendaciones de diferentes organizaciones de salud. RESULTADOS: Se incluyeron dos RS para umifenovir, un estudio observacional controlado para favipiravir, un estudio observacional controlado para favipiravir versus umifenovir, así como 13 GPC y recomendaciones de sociedades científicas. CONCLUSIONES Evidencia de muy baja calidad proveniente de estudios observacionales (publicación preliminar de revisiones sistemáticas) podría sugerir que el uso de favipiravir o umifenovir comparado con otros antivirales o atención estándar reduciría los tiempos hasta la negativización viral determinada por PCR en los pacientes con COVID-19 leves o moderados, sin encontrar diferencias en la mejoría de los síntomas a los 14 días. No evaluaron tasas de mortalidad. Evidencia de muy baja calidad proveniente de un estudio observacional controlado que compara favipiravir versus umifenovir no encontró diferencias entre ambos tratamientos en días a la recuperación, ni en la mejoría de ningún síntoma a los 7 días. No evaluaron tasas de mortalidad. Ninguna de las guías o protocolos gubernamentales o de sociedades científicas recomiendan el uso de favipiravir o umifenovir para COVID-19. Tampoco se encontraron políticas de cobertura que los mencionen. Algunos ensayos clínicos aleatorizados que evalúan la eficacia y seguridad de estas drogas en pacientes con cuadros respiratorios por COVID-19 se encuentran en curso y podrían cambiar esta evidencia.

As causas das internações no Hospital Adauto Botelho (Cariacica, ES) na segunda metade do século XX / The motives for hospitalization at Adauto Botelho Hospital (Cariacica, ES) in the second half of the twentieth century

Discutem-se aqui as formas de encaminhamento de pacientes ao Hospital Adauto Botelho, localizado em Cariacica, Espírito Santo. A pesquisa se deu por meio de prontuários médicos datados desde a inauguração em 1954 e de depoimentos de pessoas que trabalharam lá durante a segunda metade do século XX. Foram analisados 102 prontuários e entrevistadas quatro pessoas. A pesquisa dos prontuários mostra forte inserção da Chefatura de Polícia no processo de internação. As falas dos entrevistados reiteram esse ponto, mostrando também a longa duração das internações. São as histórias de vida dos internos que dão o tom deste trabalho. Conclui-se, a partir delas, que o Hospital Adauto Botelho, mais que uma instituição de tratamento, era um espaço de confinamento.

This paper discusses the procedures for referring patients to Adauto Botelho Hospital, in Cariacica, Espírito Santo state, Brazil. The research is based on the medical records since its inauguration in 1954 and statements by people who worked there in the second half of the twentieth century. One hundred and two records were analyzed and four people were interviewed. The records revealed the active involvement of the Chief of Police in hospitalizations. The interviews corroborate this, while also showing the long duration of the hospitalizations. The tone of the paper is set by the life stories of the people hospitalized there. The conclusion is that this hospital served not so much for treatment as for confinement.

The rescue of H1N1 subtype swine influenza virus / 生物工程学报

The swine influenza virus (SIV) strain A/Swine/TianJin/01/2004(H1N1) (A/S/TJ/04) was rescued successfully by an eight-plasmid rescue system. The cDNAs of SIV 8 gene segments were synthesized by RT-PCR and cloned into the RNA polymerase I/II bidirection expression vector PHW2000 independently, resulting in 8 recombinant plasmids. The 8 recombinant plasmids were cotransfected into COS-1 cell, 30 h later TPCK-trypsin was added to 0.5 microg/mL. The COS-1 cell and supernatant were harvested 48 h after cotransfection and were inoculated into the allantoic cavity of 9-day-old specific-pathogen free (SPF) chicken eggs. The allantoic fluid of dead eggs was harvested and passaged 3 generations in SPF chicken eggs to get infective virus. The successful rescue of A/S/TJ/04 SIV was identified by hemagglutination assay, hemagglutination inhibition assay, sequence analysis and electron microscope observation. The successful rescue of SIV built a platform for the research of the relationship between genome structure and function of SIV, the mechanisms of trans-species transmission of influenza virus and for the generation of new SIV as vaccine.

RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species

Leishmania (Sauroleishmania) tarentolae has biotechnological potential for use as live vaccine against visceral leishmaniasis and as a system for the over expression of eukaryotic proteins that possess accurate post-translational modifications. For both purposes, new systems for protein expression in this non-pathogenic protozoan are necessary. The ribosomal RNA promoter proved to be a stronger transcription driver since its use yielded increased levels of recombinant protein in organisms of both genera Trypanosoma or Leishmania. We have evaluated heterologous expression systems using vectors with two different polypyrimidine tracts in the splice acceptor site by measuring a reporter gene transcribed from L. tarentolae RNA polymerase I promoter. Our data indicate that the efficiency of chloramphenicol acetyl transferase expression changed drastically with homologous or heterologous sequences, depending on the polypyrimidine tract used in the construct and differences in size and/or distance from the AG dinucleotide. In relation to the promoter sequence the reporter expression was higher in heterologous lizard-infecting species than in the homologous L. tarentolae or in the mammalian-infecting L. (Leishmania) amazonensis.

A Case of Renal Crisis in a Korean Scleroderma Patient with Anti-RNA polymerase I and III Antibodies

Scleroderma (SSc) renal crisis has been reported to be associated with anti-RNA polymerase I and III (RNAP I/III) antibodies in Caucasians and the Japanese. However, no report is available for Korean SSc patients. Here, we describe the case of a 65-yr-old female SSc patient who developed renal crisis and whose serum contained anti-RNAP I/III antibodies. She was finally diagnosed as having diffuse cutaneous SSc based on skin thickening proximal to the elbows and knees. Sudden hypertension, oliguria, and pulmonary edema were features of her renal crisis. Despite the use of captopril and adequate blood pressure control, her renal function deteriorated. Subsequent renal biopsy findings showed severe fibrinoid necrosis with luminal obliteration in interlobar arteries and arterioles consistent with SSc renal crisis. Serum anti-RNAP I/III antibodies were detected by radioimmunoprecipitation. This is the first report of a renal crisis in a Korean SSc patient with RNAP I/III antibodies.

Colocalization of coilin and nucleolar proteins in Cajal body-like structures of micronucleated PtK2 cells

Cajal bodies (CB) are ubiquitous nuclear structures involved in the biogenesis of small nuclear ribonucleoproteins and show narrow association with the nucleolus. To identify possible relationships between CB and the nucleolus, the localization of coilin, a marker of CB, and of a set of nucleolar proteins was investigated in cultured PtK2 cells undergoing micronucleation. Nocodazol-induced micronucleated cells were examined by double indirect immunofluorescence with antibodies against coilin, fibrillarin, NOR-90/hUBF, RNA polymerase I, PM/Scl, and To/Th. Cells were imaged on a BioRad 1024-UV confocal system attached to a Zeiss Axiovert 100 microscope. Since PtK2 cells possess only one nucleolus organizer region, micronucleated cells presented only one or two micronuclei containing nucleolus. By confocal microscopy we showed that in most micronuclei lacking a typical nucleolus a variable number of round structures were stained by antibodies against fibrillarin, NOR-90/hUBF protein, and coilin. These bodies were regarded as CB-like structures and were not stained by anti-PM/Scl and anti-To/Th antibodies. Anti-RNA polymerase I antibodies also reacted with CB-like structures in some micronuclei lacking nucleolus. The demonstration that a set of proteins involved in RNA/RNP biogenesis, namely coilin, fibrillarin, NOR-90/hUBF, and RNA polymerase I gather in CB-like structures present in nucleoli-devoid micronuclei may contribute to shed some light into the understanding of CB function.

The Relationship of AgNORs to the Survival Rate of Patients with Invasive

Nucleolar organizer regions(NORs) are argyrophilic and have a loops of DNA that tran scribes to ribosomal RNA by RNA Polymerase I and are involved in formation of nucleolus. They are used in measuring proliferative activity of cells. Mean AgNORs count is increased or large and NORs are irregular in shape in malignant tumors. To examine the relationship of AgNORs to the survival rate of the patients with invasive colorectal adenocarcinoma, we counted mean AgNORs in 200 tumor cells and mean AgNORs#(mean AgNORs in counting AgNORs of the upper 40 nuclei after sorting AgNORs counts of 200 cell nuclei in descending order) after AgNOR staining using 45 cases of formalin-fixed, paraffin-embedded tissue surgically excised at Kang-Dong Sacred heart hospital. The survival rates were not related to age, sex, locations, differentiation, mAgNORs and mAgNORs# except Dukes'stages. There were only statistical significances in AgNORs# between high and low age group(p=0.0001), and between colon and rectum(p=0.005). But it is known that there could be different results according to staining time, fixation and preservation of tissues, counting methods in measuring AgNORs. There might be clinical usefulness of AgNOR if more cases are examined together with better tissue processing and staining methods.

Analysis of rpoB Gene in Rifampin-Resistant M. Tuberculosis by Direct Sequencing and Line Probe Assay / 결핵

BACKGROUND: The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in beta subunit of RNA polymerase is main mechanism of resistance. METHOD: In this study, rpoB gene for the ~3 subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. RESULTS: All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S53 IL) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not comfirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513-515 were new identified mutations for the first time. CONCLUSION: Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.

The significance of the nucleolar organizer regions(NORs) in prostatic adenocarcinoma / 대한비뇨기과학회지

NORs are loops of DNA which occur in nucleoli and which posses ribosomal DNA genes. Ribosomal DNA genes transcribe to ribosomal RNA by RNA polymerase I and are of vital significance in the ultimate synthesis of protein. Proteins associated with the NORs are stained with silver nitrate (Ag-NORs). Ag-NORs were studied in various tumors and might reflect the activity of cells and might be an indicator of the degree of malignancy in tumors. Specimens of prostate obtained by transurethral resection or fine needle biopsy were examined with regard to the number and area of NORs in order to find out whether the above mentioned two parameters reflect proliferative activity in 23 cases of prostatic adenocarcinoma and 10 cases of benign prostatic hyperplasia as controls. The mean number of Ag-NORs in BPH, well, moderate and poorly differentiated adenocarcinoma was 1.24+/-0.10 (mean+/-SD; n=10), 1.87+/-0.23 (n=6). 2.08 J=0.28 (n=10). 4.02+/-0.22 (n=7). respectively. In each group, significant difference was found statistically (p<0.05) except between well and moderate differentiated adenocarcinoma. The mean area of Ag-NORs calculated by morphometry in BPH, well, moderate and poorly differentiated adenocarcinoma was 5.40+/-1.8 um2, 7.84+/-3.68 um2. 14.02+/-4.95 um2, 23.99+/-6.94 um2, respectively and significant difference was found statistically in each group (p<0.05). We concluded that the number and area of NORs increased with degree of malignancy and may reflect proliferative activity of cells in prostatic adenocarcinoma. Further investigations may be needed to confirm whether the number or area of Ag-NORs may be available as a prognostic factor for patient`s survival.

The significance of nucleolar organizer regions in bladder epithelial lesions induced by N-butyl-N-(4-hydroxybutyl)nitrosamine in rat / 대한비뇨기과학회지

It is known that the activity of tumor cells correlates with the malignant potential of tumors. Nucleolar organizer regions(NORs) are loops of DNA which occur in nucleoli and which possess ribosomal DNA genes. Ribosomal DNA genes transcribe to ribosomal RNA by RNA polymerase I and are of vital significance in the ultimate synthesis of protein. Proteins associated with the NORs are stained with silver nitrate(Ag-NORs). Ag-NORs were studied in various tumors and might reflect the activity of cells and might be and indicator of the degree of malignancy in tumors. 0.05 % N-Butyl-N(4-hydroxybutyl) nitrosamine(BBN) in drinking water was given for 8 and 14 weeks to induce pre-neoplastic and neoplastic vesical epithelial lesions in female Wister rats. Ag-NORs were stained by the simple one-step silver colloid staining in routine processed, formalin-fixed paraffin sections of bladder lesions induced by N-butyl-N(4-hydroxybutyl) nitrosamine(BBN) in rat. The mean number of silver stained NORs(Ag-NORs) was as follows : normal transitional epithelium was 1.64+/-0.25(mean+/-SD; n=10), simple hyperplasia was 2.24+/-0.24(n=7). nodular hyperplasia was 2.63+/-0.12 (n=6), transitional cell papilloma was 3.24+/-0.28(n=6) and transitional cell carcinoma was 4.52+/-0.32(n=10). Based on the above results, we concluded that the mean number of Ag-NORs showed a stepwise increase from normal transitional epithelium through simple hyperplasia and nodular hyperplasia to papilloma and carcinoma of bladder epithelium induced by BBN in rat and may reflect proliferative activity of cells. Further investigations may be needed to confirm whether the number or quantification of Ag-NORs is a useful method for evaluating proliferative activity of neoplastic lesions or another indicator of prognosis in human bladder cancer.

Nucleolar organizer regions in glioma / 영남의대학술지

Nucleolar organizer regions (NOR) are loops of ribosomal DNA (rDNA) which are transcribed by RNA polymerase I. They produce ultimately ribosome and protein. Thus they are believed to reflect nuclear activity. We applied silver colloid staining technique to human glioma to examine relationship between the mean number of Ag-NOR and histopathological grading. The mean number of Ag-NOR (±S. E of the mean) were 1.17±0.07 in normal brain, 1.53±0.25 in astrocytoma, 2.37±0.71 in malignant astrocytoma. And 2.88±0.41 in glioblastoma multiforme. And there was a statistically significant difference among these. The results show that Ag-NOR technique is a rather simple and rapid method and will become a helpful tool for estimation of the proliferative potential of glioma.