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BACKGROUND: Governments in Latin America are constantly facing the problem of managing scarce resources to satisfy alternative needs, such as housing, education, food, and healthcare security. Those needs, combined with increasing crime levels, require financial resources to be solved. OBJECTIVE: The objective of this review was to characterizar the health system and health expenditure of a large country (Brazil) and a small country (Chile) and identify some of the challenges these two countries face in improving the health services of their population. METHODS: A literature review was conducted by searching journals, databases, and other electronic resources to identify articles and research publications describing health systems in Brazil and Chile. RESULTS: The review showed that the economic restriction and the economic cycle have an impact on the funding of the public health system. This result was true for the Brazilian health system after 2016, despite the change to a unique health system one decade earlier. In the case of Chile, there are different positions about which one is the best health system: a dual public and private or just public one. As a result, a referendum on September 4, 2022, of a new constitution, which incorporated a unique health system, was rejected. At the same time, the Government ended the copayment in the public health system in September 2022, excluding illnesses referred to the private sector. Another issue detected was the fragility of the public and private sector coverage due to the lack of funding. CONCLUSIONS: The health care system in Chile and Brazil has improved in the last decades. However, the public healthcare systems still need additional funding and efficiency improvement to respond to the growing health requirements needed from the population.
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Autosomal dominant polycystic kidney disease (ADPKD), primarily due to PKD1 or PKD2 mutations, causes progressive kidney cyst development and kidney failure. There is significant intrafamilial variability likely due to the genetic background and environmental/lifestyle factors; variability that can be modeled in PKD mice. Here, we characterized mice homozygous for the PKD1 hypomorphic allele, p.Arg3277Cys (Pkd1RC/RC), inbred into the BALB/cJ (BC) or the 129S6/SvEvTac (129) strains, plus F1 progeny bred with the previously characterized C57BL/6J (B6) model; F1(BC/B6) or F1(129/B6). By one-month cystic disease in both the BC and 129 Pkd1RC/RC mice was more severe than in B6 and continued with more rapid progression to six to nine months. Thereafter, the expansive disease stage plateaued/declined, coinciding with increased fibrosis and a clear decline in kidney function. Greater severity correlated with more inter-animal and inter-kidney disease variability, especially in the 129-line. Both F1 combinations had intermediate disease severity, more similar to B6 but progressive from one-month of age. Mild biliary dysgenesis, and an early switch from proximal tubule to collecting duct cysts, was seen in all backgrounds. Preclinical testing with a positive control, tolvaptan, employed the F1(129/B6)-Pkd1RC/RC line, which has moderately progressive disease and limited isogenic variability. Magnetic resonance imaging was utilized to randomize animals and provide total kidney volume endpoints; complementing more traditional data. Thus, we show how genetic background can tailor the Pkd1RC/RC model to address different aspects of pathogenesis and disease modification, and describe a possible standardized protocol for preclinical testing.
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Rim Policístico Autossômico Dominante , Animais , Patrimônio Genético , Rim , Camundongos , Camundongos Endogâmicos C57BL , Mutação , Rim Policístico Autossômico Dominante/genética , Canais de Cátion TRPP/genéticaRESUMO
Autosomal-dominant polycystic kidney disease (ADPKD) is characterized by the progressive development of kidney cysts, often resulting in end-stage renal disease (ESRD). This disorder is genetically heterogeneous with â¼7% of families genetically unresolved. We performed whole-exome sequencing (WES) in two multiplex ADPKD-like pedigrees, and we analyzed a further 591 genetically unresolved, phenotypically similar families by targeted next-generation sequencing of 65 candidate genes. WES identified a DNAJB11 missense variant (p.Pro54Arg) in two family members presenting with non-enlarged polycystic kidneys and a frameshifting change (c.166_167insTT) in a second family with small renal and liver cysts. DNAJB11 is a co-factor of BiP, a key chaperone in the endoplasmic reticulum controlling folding, trafficking, and degradation of secreted and membrane proteins. Five additional multigenerational families carrying DNAJB11 mutations were identified by the targeted analysis. The clinical phenotype was consistent in the 23 affected members, with non-enlarged cystic kidneys that often evolved to kidney atrophy; 7 subjects reached ESRD from 59 to 89 years. The lack of kidney enlargement, histologically evident interstitial fibrosis in non-cystic parenchyma, and recurring episodes of gout (one family) suggested partial phenotypic overlap with autosomal-dominant tubulointerstitial diseases (ADTKD). Characterization of DNAJB11-null cells and kidney samples from affected individuals revealed a pathogenesis associated with maturation and trafficking defects involving the ADPKD protein, PC1, and ADTKD proteins, such as UMOD. DNAJB11-associated disease is a phenotypic hybrid of ADPKD and ADTKD, characterized by normal-sized cystic kidneys and progressive interstitial fibrosis resulting in late-onset ESRD.
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Alelos , Proteínas de Choque Térmico HSP40/genética , Mutação/genética , Rim Policístico Autossômico Dominante/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Sequência de Bases , Células Epiteliais/metabolismo , Família , Feminino , Proteínas de Choque Térmico HSP40/química , Humanos , Alça do Néfron/patologia , Masculino , Pessoa de Meia-Idade , Linhagem , Rim Policístico Autossômico Dominante/diagnóstico por imagem , Rim Policístico Autossômico Dominante/patologia , Canais de Cátion TRPP/genética , Uromodulina/metabolismo , Sequenciamento do Exoma , Adulto JovemRESUMO
Vascular abnormalities are the most important non-cystic complications in Polycystic Kidney Disease (PKD) and contribute to renal disease progression. Endothelial dysfunction and oxidative stress are evident in patients with ADPKD, preserved renal function, and controlled hypertension. The underlying biological mechanisms remain unknown. We hypothesized that in early ADPKD, the reactive oxygen species (ROS)-producing nicotinamide adenine dinucleotide phosphate hydrogen (NAD(P)H)-oxidase complex-4 (NOX4), a major source of ROS in renal tubular epithelial cells (TECs) and endothelial cells (ECs), induces EC mitochondrial abnormalities, contributing to endothelial dysfunction, vascular abnormalities, and renal disease progression. Renal oxidative stress, mitochondrial morphology (electron microscopy), and NOX4 expression were assessed in 4- and 12-week-old PCK and Sprague-Dawley (wild-type, WT) control rats (n = 8 males and 8 females each). Endothelial function was assessed by renal expression of endothelial nitric oxide synthase (eNOS). Peritubular capillaries were counted in hematoxylin-eosin (H&E)-stained slides and correlated with the cystic index. The enlarged cystic kidneys of PCK rats exhibited significant accumulation of 8-hydroxyguanosine (8-OHdG) as early as 4 weeks of age, which became more pronounced at 12 weeks. Mitochondria of TECs lining cysts and ECs exhibited loss of cristae but remained preserved in non-cystic TECs. Renal expression of NOX4 was upregulated in TECs and ECs of PCK rats at 4 weeks of age and further increased at 12 weeks. Contrarily, eNOS immunoreactivity was lower in PCK vs. WT rats at 4 weeks and further decreased at 12 weeks. The peritubular capillary index was lower in PCK vs. WT rats at 12 weeks and correlated inversely with the cystic index. Early PKD is associated with NOX4-induced oxidative stress and mitochondrial abnormalities predominantly in ECs and TECs lining cysts. Endothelial dysfunction precedes capillary loss, and the latter correlates with worsening of renal disease. These observations position NOX4 and EC mitochondria as potential therapeutic targets in PKD.
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Rim/patologia , Mitocôndrias/patologia , NADPH Oxidase 4/metabolismo , Óxido Nítrico Sintase Tipo III/metabolismo , Doenças Renais Policísticas/patologia , Animais , Progressão da Doença , Células Endoteliais/metabolismo , Células Epiteliais/metabolismo , Feminino , Guanosina/análogos & derivados , Guanosina/análise , Masculino , Óxido Nítrico Sintase Tipo III/biossíntese , Estresse Oxidativo/fisiologia , Ratos , Ratos Sprague-Dawley , Espécies Reativas de Oxigênio/metabolismoRESUMO
ß-Adrenergic receptor blockade reduces total mortality and all-cause hospitalizations in patients with heart failure (HF). Nonetheless, ß-blockade does not halt disease progression, suggesting that cAMP-dependent protein kinase (PKA) signaling downstream of ß-adrenergic receptor activation may persist through unique post-translational states. In this study, human myocardial tissue was used to examine the state of PKA subunits. As expected, total myosin binding protein-C phosphorylation and Ser23/24 troponin I phosphorylation significantly decreased in HF. Examination of PKA subunits demonstrated no change in type II regulatory (RIIα) or catalytic (Cα) subunit expression, although site specific RIIα (Ser96) and Cα (Thr197) phosphorylation were increased in HF. Further, the expression of type I regulatory subunit (RI) was increased in HF. Isoelectric focusing of RIα demonstrated up to three variants, consistent with reports that Ser77 and Ser83 are in vivo phosphorylation sites. Western blots with site-specific monoclonal antibodies showed increased Ser83 phosphorylation in HF. 8-fluo-cAMP binding by wild type and phosphomimic Ser77 and Ser83 mutant RIα proteins demonstrated reduced Kd for the double mutant as compared to WT RIα. Therefore, failing myocardium displays altered expression and post-translational modification of PKA subunits that may impact downstream signaling.
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Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Insuficiência Cardíaca/diagnóstico , Insuficiência Cardíaca/metabolismo , Processamento de Proteína Pós-Traducional , Subunidades Proteicas/metabolismo , Antagonistas Adrenérgicos beta/farmacologia , Adulto , Idoso , Domínio Catalítico , AMP Cíclico/metabolismo , Feminino , Coração/fisiologia , Holoenzimas/química , Humanos , Masculino , Pessoa de Meia-Idade , Miocárdio/metabolismo , Fosforilação , Serina/química , Troponina I/químicaRESUMO
Introduction: Dent disease is an X-linked recessive disorder associated with low molecular weight proteinuria (LMWP), nephrocalcinosis, kidney stones, and kidney failure in the third to fifth decade of life. It consists of Dent disease 1 (DD1) (60% of patients) because of pathogenic variants in the CLCN5 gene and Dent disease 2 (DD2) with changes in OCRL. Methods: Retrospective review of 162 patients from 121 different families with genetically confirmed DD1 (82 different pathogenic variants validated using American College of Medical Genetics [ACMG] guidelines). Clinical and genetic factors were compared using observational statistics. Results: A total of 110 patients had 51 different truncating (nonsense, frameshifting, large deletions, and canonical splicing) variants, whereas 52 patients had 31 different nontruncating (missense, in-frame, noncanonical splicing, and stop-loss) changes. Sixteen newly described pathogenic variants were found in our cohort. Among patients with truncating variants, lifetime stone events positively correlated with chronic kidney disease (CKD) evolution. Patients with truncating changes also experienced stone events earlier in life and manifested a higher albumin excretion rate than the nontruncating group. Nevertheless, neither age of nephrocalcinosis nor CKD progression varied between the truncating versus nontruncating patients. A large majority of nontruncating changes (26/31; 84%) were clustered in the middle exons that encode the voltage ClC domain whereas truncating changes were spread across the protein. Variants associated with kidney failure were restricted to truncating (11/13 cases), plus a single missense variant previously shown to markedly reduce ClC-5 functional activity that was found in the other 2 individuals. Conclusion: DD1 manifestations, including the risk of kidney stones and progression to kidney failure, may relate to the degree of residual ClC-5 function.
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Rationale & Objective: Hereditary hypophosphatemic rickets with hypercalciuria (HHRH) is a rare monogenic disorder caused by SLC34A3 pathogenic variants. HHRH is characterized by kidney phosphate wasting, hypophosphatemia, hypercalciuria, an elevated 1,25-dihydroxyvitamin D level, nephrocalcinosis, and urinary stone disease. Previously, we reported a 100% prevalence of kidney cysts in the related CYP24A1 deficiency. Thus, in the current study, we characterized cysts' presence in HHRH, another monogenic cause of hypercalciuria, nephrocalcinosis, and urinary stone disease. Study Design: Case series. Setting & Participants: Medical records from the Mayo Clinic and the Rare Kidney Stone Consortium monogenic stone disease database were queried for patients with a genetically confirmed HHRH diagnosis. The number, sizes, and locations of kidney cysts in each patient were recorded. Results: Twelve patients with SLC34A3 pathogenic variants were identified (7 monoallelic, 5 biallelic). Of these, 5 (42%) were males, and the median (Q1, Q3) ages were 16 years (13, 35 years) at clinical presentation and 42 years (20, 57 years) at genetic confirmation. Kidney cysts were present in 9 of 12 (75%) patients, and the median (Q1, Q3) age at first cyst detection was 41 years (13, 50 years). The median number of cysts per patient was 2.0 (0.5, 3.5). Fifty percent of adult patients had a cyst number that exceeded the 97.5th percentile of an age- and sex-matched control population. All children had at least 2 or more total cysts. None had a family history of cystic kidney disease. Limitations: Retrospective study, possible selection bias, single-center experience. Conclusions: A strong association between HHRH and kidney cysts was observed. Similarities in the biochemical profiles of HHRH and CYP24A1 deficiency suggest elevated active vitamin D and hypercalciuria may be potential cystogenic factors. Further studies are needed to understand how genetic changes in SLC34A3 favor cyst formation.
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Dent disease is a rare X-linked renal tubulopathy due to CLCN5 and OCRL (DD2) mutations. OCRL mutations also cause Lowe syndrome (LS) involving the eyes, brain and kidney. DD2 is frequently described as a mild form of LS because some patients may present with extra-renal symptoms (ESs). Since DD2 is a rare disease and there are a low number of reported cases, it is still unclear whether it has a clinical picture distinct from LS. We retrospectively analyzed the phenotype and genotype of our cohort of 35 DD2 males and reviewed all published DD2 cases. We analyzed the distribution of mutations along the OCRL gene and evaluated the type and frequency of ES according to the type of mutation and localization in OCRL protein domains. The frequency of patients with at least one ES was 39%. Muscle findings are the most common ES (52%), while ocular findings are less common (11%). Analysis of the distribution of mutations revealed (1) truncating mutations map in the PH and linker domain, while missense mutations map in the 5-phosphatase domain, and only occasionally in the ASH-RhoGAP module; (2) five OCRL mutations cause both DD2 and LS phenotypes; (3) codon 318 is a DD2 mutational hot spot; (4) a correlation was found between the presence of ES and the position of the mutations along OCRL domains. DD2 is distinct from LS. The mutation site and the mutation type largely determine the DD2 phenotype.
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Doenças Genéticas Ligadas ao Cromossomo X/genética , Pleiotropia Genética/genética , Nefrolitíase/genética , Síndrome Oculocerebrorrenal/genética , Monoéster Fosfórico Hidrolases/genética , Adolescente , Variação Biológica da População/genética , Criança , Pré-Escolar , Feminino , Estudos de Associação Genética , Doenças Genéticas Ligadas ao Cromossomo X/diagnóstico , Doenças Genéticas Ligadas ao Cromossomo X/epidemiologia , Genótipo , Humanos , Rim/metabolismo , Rim/patologia , Masculino , Mutação de Sentido Incorreto/genética , Nefrolitíase/diagnóstico , Nefrolitíase/epidemiologia , Síndrome Oculocerebrorrenal/diagnóstico , Síndrome Oculocerebrorrenal/epidemiologia , FenótipoRESUMO
INTRODUCTION: Because of phenotypic overlap between monogenic urinary stone diseases (USD), gene-specific analyses can result in missed diagnoses. We used targeted next generation sequencing (tNGS), including known and candidate monogenic USD genes, to analyze suspected primary hyperoxaluria (PH) or Dent disease (DD) patients genetically unresolved (negative; N) after Sanger analysis of the known genes. Cohorts consisted of 285 PH (PHN) and 59 DD (DDN) families. METHODS: Variants were assessed using disease-specific and population databases plus variant assessment tools and categorized using the American College of Medical Genetics (ACMG) guidelines. Prior Sanger analysis identified 47 novel PH or DD gene pathogenic variants. RESULTS: Screening by tNGS revealed pathogenic variants in 14 known monogenic USD genes, accounting for 45 families (13.1%), 27 biallelic and 18 monoallelic, including 1 family with a copy number variant (CNV). Recurrent genes included the following: SLC34A3 (n = 13), CLDN16 (n = 8), CYP24A1 (n = 4), SLC34A1 (n = 3), SLC4A1 (n = 3), APRT (n = 2), CLDN19 (n = 2), HNF4A1 (n = 2), and KCNJ1 (n = 2), whereas ATP6V1B1, CASR, and SLC12A1 and missed CNVs in the PH genes AGXT and GRHPR accounted for 1 pedigree each. Of the 48 defined pathogenic variants, 27.1% were truncating and 39.6% were novel. Most patients were diagnosed before 18 years of age (76.1%), and 70.3% of biallelic patients were homozygous, mainly from consanguineous families. CONCLUSION: Overall, in patients suspected of DD or PH, 23.9% and 7.3% of cases, respectively, were caused by pathogenic variants in other genes. This study shows the value of a tNGS screening approach to increase the diagnosis of monogenic USD, which can optimize therapies and facilitate enrollment in clinical trials.
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COVID-19 , SARS-CoV-2 , Anticorpos Neutralizantes , Anticorpos Antivirais , COVID-19/diagnóstico , HumanosRESUMO
Introducción: La pérdida del anillo neurorretiniano (ANR) es una característica distintiva del glaucoma que genera desviación de la regla ISNT. Objetivos: Determinar el porcentaje de ojos sanos que cumplen la regla ISNT y establecer cuál de sus variantes es más aplicable a la población normal. Métodos: Evaluación del orden del grosor del anillo neurorretiniano por medio de fotografía de disco óptico, capa de fibras nerviosas en tomografía de coherencia óptica (OCT) y evaluación clínica por oftalmoscopia indirecta. Resultados: Se evaluaron 102 ojos sanos de pacientes con edad promedio 44.1 años. El porcentaje de la regla ISNT en foto de disco, OCT y oftalmoscopia indirecta se cumplió en el 36.3, 38.2 y 29.4% de los ojos respectivamente, la variación de la regla en la que se excluye cuadrante nasal y temporal, conocida como regla «IS¼, aumentó sus porcentajes de cumplimiento al 73.5, 52.9 y 54.9% al ser evaluadas en foto de disco, OCT y oftalmoscopia indirecta, respectivamente. Conclusiones: La regla ISNT solo es válida en un tercio de los ojos evaluados mediante los métodos descrito
Background: Neurorretinal rim loss is an important characteristic of glaucoma that generates deviation from the ISNT rule. Objective: To determine what percentage of normal eyes follow the ISNT rule, and wich ISNT rule variants may be more generalizable to the normal population. Methods: Neurorretinal rim assessment from optic disc photographs, retinal nerve fiber layer (RNFL) thickness measurements from (OCT) and clinical evaluation by indirect ophthalmoscopy. Results: 102 healthy eyes were evaluated, with an average age of 44.1 ± 11.7 years. The percentage of agreement of ISNT rule was verified in in disk photo (36.3%), OCT (38.2%) and indirect ophthalmoscopy (29.4%) of the total population. The variation of the rule in which the nasal (73.5) and temporal (52.9%) quadrant are excluded, known as the "IS" rule, increased its compliance percentages from 73.5%, 52.9% and 54.9% when evaluated in disk photo, OCT and indirect ophthalmoscopy, respectively.
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Humanos , Masculino , FemininoRESUMO
Introducción: Las estructuras ópticas pueden variar en su configuración y medida dependiendo de la edad, el sexo y la raza, por lo que su medición previa a la cirugía es un pilar fundamental para el cálculo correcto del lente intraocular. Caracterizar los rangos normales de estos parámetros en nuestra población suministra información importante que puede mejorar los resultados visuales tras la cirugía de catarata y alertar en aquellos casos que distan de la media de los rangos conocidos. Objetivo: Describir los parámetros biométricos oculares en pacientes candidatos a cirugía de catarata y analizar sus variaciones según la edad y el sexo. Método: Estudio observacional descriptivo, de corte transversal. Análisis retrospectivo de biometrías oculares realizadas con reflectometría óptica de baja coherencia en pacientes candidatos a cirugía de catarata. Resultados: Se evaluaron 820 ojos de 820 pacientes candidatos a cirugía de catarata, con una edad media de 68.1 ± 11.1 años, el 53.7% de sexo femenino. Los parámetros oculares promedios encontrados fueron: longitud axial (AL) 23.39 ± 1.11 mm, profundidad de cámara anterior (ACD) 3.37 ± 0.49 mm, astigmatismo corneal (AST) 1.09 ± 0.85 D, espesor corneal central (CCT) 529.43 ± 35.02 µm, poder queratométrico (K) 43.81 ± 1.64 D, queratometría plana 43.28 ± 1.69 D, queratometría curva 44.37 ± 1.74 D y distancia blanco a blanco 11.82 ± 0.52 mm. El promedio de AL y ACD fue mayor en los hombres que en las mujeres (p < 0.0001), y con el aumento de la edad el AST y el K mostraron una tendencia al incremento. Se encontró un 23.78% de astigmatismo con la regla, un 54.88% de astigmatismo contra la regla y un 21.34% de astigmatismo oblicuo. Conclusiones: El perfil de los datos biométricos evidenció diferencias significativas entre sexos y grupos de edad. Se encontró una buena correlación entre la AL, la ACD y la potencia corneal, y los promedios encontrados se corresponden bien con los de otras poblaciones de similar origen étnico.
Introduction: Optical structures can vary in their configuration and measurement depending on age, sex and race, so their measurement, prior to surgery, is a fundamental pillar for the correct calculation of the intraocular lens. Characterizing the normal ranges of these parameters in our population provides important information that can improve the visual results of cataract surgery and alert in those cases that are far from the mean of the known ranges. Objective: To describe the ocular biometric parameters in cataract surgery candidates. Method: Descriptive, cross-sectional study. Retrospective analysis of biometric parameters measured by optical low-coherence reflectometry in cataract surgery candidates. Results: The study evaluated 820 eyes of 820 cataract patients, the mean age was 68.1 ± 11.1 years, 53.7% female. Mean ocular parameters found were: axial length (AL) 23.39 ± 1.11 mm, anterior chamber depth (ACD) 3.37 ± 0.49 mm, corneal astigmatism (AST) 1.09 ± 0.85 D, central corneal thickness (CCT) 529.43 ± 35.02 µm, keratometry (K) 43.81 ± 1.64 D, flat keratometry (K1) 43.28 ± 1.69 D, steep keratometry (K2) 44.37 ± 1.74 D and white-to-white distance (WTW) 11.82 ± 0.52 mm. Mean AL and ACD were significantly longer in men (p < 0.0001) and with increasing age, AST and K showed an upward trend. 23.78% of astigmatism with the rule, 54.88% of astigmatism against the rule and 21.34% of oblique astigmatism were found. Conclusions: The profile of the biometric data showed significant differences between sex and age groups, a good correlation was found between axial length, chamber depth and keratometry. The averages found correspond well to those of other populations of similar ethnic origin
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Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , BiometriaRESUMO
BACKGROUND: Human influenza virus isolates generally grow poorly in embryonated chicken eggs. Hence, gene reassortment of influenza A wild type (wt) viruses is performed with a highly egg adapted donor virus, A/Puerto Rico/8/1934 (PR8), to provide the high yield reassortant (HYR) viral 'seeds' for vaccine production. HYR must contain the hemagglutinin (HA) and neuraminidase (NA) genes of wt virus and one to six 'internal' genes from PR8. Most studies of influenza wt and HYRs have focused on the HA gene. The main objective of this study is the identification of the molecular signature in all eight gene segments of influenza A HYR candidate vaccine seeds associated with high growth in ovo. METHODOLOGY: The genomes of 14 wt parental viruses, 23 HYRs (5 H1N1; 2, 1976 H1N1-SOIV; 2, 2009 H1N1pdm; 2 H2N2 and 12 H3N2) and PR8 were sequenced using the high-throughput sequencing pipeline with big dye terminator chemistry. RESULTS: Silent and coding mutations were found in all internal genes derived from PR8 with the exception of the M gene. The M gene derived from PR8 was invariant in all 23 HYRs underlining the critical role of PR8 M in high yield phenotype. None of the wt virus derived internal genes had any silent change(s) except the PB1 gene in X-157. The highest number of recurrent silent and coding mutations was found in NS. With respect to the surface antigens, the majority of HYRs had coding mutations in HA; only 2 HYRs had coding mutations in NA. SIGNIFICANCE: In the era of application of reverse genetics to alter influenza A virus genomes, the mutations identified in the HYR gene segments associated with high growth in ovo may be of great practical benefit to modify PR8 and/or wt virus gene sequences for improved growth of vaccine 'seed' viruses.
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Genoma Viral/genética , Vírus da Influenza A/genética , Hemaglutininas/genética , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/genética , Vírus da Influenza A Subtipo H3N2/imunologia , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/imunologia , Vírus da Influenza A/imunologia , Vacinas contra Influenza/genética , Vacinas contra Influenza/imunologia , Mutação , Neuraminidase/genéticaRESUMO
BACKGROUND: Influenza A virus vaccines undergo yearly reformulations due to the antigenic variability of the virus caused by antigenic drift and shift. It is critical to the vaccine manufacturing process to obtain influenza A seed virus that is antigenically identical to circulating wild type (wt) virus and grows to high titers in embryonated chicken eggs. Inactivated influenza A seasonal vaccines are generated by classical reassortment. The classical method takes advantage of the ability of the influenza virus to reassort based on the segmented nature of its genome. In ovo co-inoculation of a high growth or yield (hy) donor virus and a low yield wt virus with antibody selection against the donor surface antigens results in progeny viruses that grow to high titers in ovo with wt origin hemagglutinin (HA) and neuraminidase (NA) glycoproteins. In this report we determined the parental origin of the remaining six genes encoding the internal proteins that contribute to the hy phenotype in ovo. METHODOLOGY: The genetic analysis was conducted using reverse transcription-polymerase chain reaction (RT-PCR) and restriction fragment length polymorphism (RFLP). The characterization was conducted to determine the parental origin of the gene segments (hy donor virus or wt virus), gene segment ratios and constellations. Fold increase in growth of reassortant viruses compared to respective parent wt viruses was determined by hemagglutination assay titers. SIGNIFICANCE: In this study fifty-seven influenza A vaccine candidate reassortants were analyzed for the presence or absence of correlations between specific gene segment ratios, gene constellations and hy reassortant phenotype. We found two gene ratios, 6:2 and 5:3, to be the most prevalent among the hy reassortants analyzed, although other gene ratios also conferred hy in certain reassortants.
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Genes Virais/genética , Vírus da Influenza A/crescimento & desenvolvimento , Vírus da Influenza A/genética , Vacinas contra Influenza/biossíntese , Vírus Reordenados/crescimento & desenvolvimento , Vírus Reordenados/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/imunologia , Humanos , Vírus da Influenza A/imunologia , Fenótipo , Polimorfismo de Fragmento de Restrição , Vírus Reordenados/imunologia , Mapeamento por RestriçãoRESUMO
The American Society for Virology 29th Annual Meeting, held in Bozeman, MT, USA, included topics covering new vaccine technologies, delivery methods and treatments in the field of virology. This conference report highlights selected presentations on human norovirus (HuNoV), SARS coronavirus and Rift Valley fever virus vaccine technologies; programmed cell death-1 (PD1) blockade and HyperAcute alpha-Gal platform technology methods; aerosol vaccination delivery; novel technologies to produce influenza virus-like particles (VLP) in mammalian cell lines; and investigational human rotavirus vaccines.
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Drogas em Investigação/administração & dosagem , Vacinas Virais/administração & dosagem , Viroses/prevenção & controle , Administração por Inalação , Aerossóis , Animais , Humanos , Tecnologia Farmacêutica/métodos , Vacinas Virais/imunologia , Viroses/imunologiaRESUMO
The H1N1 Swine Flu: The 2010 Perspective conference, held in New York City, included topics covering new research developments regarding the H1N1 influenza virus. This conference report highlights selected presentations on high-yield reassortant viral production, virus transmission and pathogenesis in ferret and guinea pig models, and the advantages of virus-like particle vaccines. Fatal pathology findings from the 2009 H1N1 strain in New York, and preparedness for and response to the 2009 pandemic, are also discussed.
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Doenças Transmissíveis Emergentes/prevenção & controle , Surtos de Doenças/prevenção & controle , Vírus da Influenza A Subtipo H1N1 , Influenza Humana/prevenção & controle , Animais , Centers for Disease Control and Prevention, U.S. , Doenças Transmissíveis Emergentes/etiologia , Doenças Transmissíveis Emergentes/patologia , Doenças Transmissíveis Emergentes/transmissão , Modelos Animais de Doenças , Indústria Farmacêutica , Furões , Cobaias , Humanos , Vírus da Influenza A Subtipo H1N1/crescimento & desenvolvimento , Vírus da Influenza A Subtipo H1N1/imunologia , Influenza Humana/etiologia , Influenza Humana/patologia , Influenza Humana/transmissão , Infecções por Orthomyxoviridae/etiologia , Infecções por Orthomyxoviridae/patologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/transmissão , Vírus Reordenados/crescimento & desenvolvimento , Tecnologia Farmacêutica/métodos , Estados Unidos , Vírion/imunologiaRESUMO
In May 2009, as the H1N1 swine flu outbreak was in the early stages, a conference was held at the New York Academy of Sciences to discuss what was known about the virus and what was being done to stop the outbreak. In May 2010, a follow-up conference was again held at the New York Academy of Sciences, but now to discuss the H1N1 outbreak retrospectively. The report presented here summarizes the 2010 conference proceedings.