Clinical and molecular characterization of Chilean patients with X-linked hypophosphatemia.

We report the most comprehensive clinical and molecular characterization of XLH patients performed in Chile. We show high prevalence of musculoskeletal burden and pain, associated with significantly impaired physical capacity and quality of life, with many relevant complications presenting more frequently than previously reported in cohorts from developed countries. INTRODUCTION: Our current understanding of the clinical presentation and natural history of X-linked hypophosphatemia (XLH) comes mainly from cohorts from developed countries, with limited data on the clinical and genetic abnormalities of XLH patients in South America. OBJECTIVE: To describe the clinical, biochemical, and molecular presentation of patients with XLH in Chile. METHODS: Patients with XLH referred by endocrinologist throughout Chile were included. Demographic data and clinical presentation were obtained from a clinical interview. Surveys were applied for quality of life (QoL), pain, and functionality. FGF23 was measured by ELISA, and genetic testing was performed. Imaging studies were conducted to assess skeletal and renal involvement. RESULTS: We included 26 patients, aged 2-64 years, from 17 unrelated Chilean families. All pediatric patients but only 40% of adults were receiving conventional therapy, while 65% of all patients had elevated alkaline phosphatase. All patients had mutations in PHEX, including 5 novel variants. Radiographic skeletal events (RSE) and enthesopathies in adults were frequent (34% and 85%, respectively). The duration of treatment was associated with fewer RSE (p < 0.05). Most adults reported pain and impaired QoL, and 50% had impaired physical capacity. The number of enthesopathies was associated with worse pain and stiffness scores (p < 0.05). CONCLUSION: Chilean patients with XLH have a high prevalence of musculoskeletal burden associated with pain and impaired physical capacity and QoL, especially in adults who were generally undertreated. These data identify a significant unmet need, inform our understanding of the current status of patients, and can guide care for XLH patients in similarly socioeconomically defined countries.

Quantitative proteomic analysis in symptomatic and asymptomatic apical periodontitis.

AIM: To quantitatively and qualitatively compare the host proteomic profile in samples of symptomatic and asymptomatic apical periodontitis (AP) using nano-liquid chromatography-electron spray tandem mass spectrometry. METHODOLOGY: Samples were obtained from 18 patients with radiographically evident AP, divided into symptomatic and asymptomatic groups (nine per group) according to clinical characteristics. After sample collection, protein extraction, purification and quantification of the samples were performed, which were analysed by reverse-phase liquid chromatography coupled to mass spectrometry. Label-free quantitative proteomic analysis was performed by Protein Lynx Global Service software. Differences in expression of proteins between the groups were calculated using the Monte Carlo algorithm, considering P < 0.05 for down-regulated proteins and 1 - P > 0.95 for up-regulated proteins. Proteins were identified with the embedded ion accounting algorithm in the software and a search of the Homo sapiens UniProt database. RESULTS: A total of 853 individual human proteins were identified. In the quantitative analysis, common proteins to both groups accounted for 143 proteins. Differences in expression between groups resulted in 51 up-regulated proteins (1 - P > 0.95) in the symptomatic group, including alpha-1-antitrypsin, protein S100-A8, myeloperoxidase, peroxiredoxin and lactotransferrin. This group also had 43 down-regulated proteins (P < 0.05), comprising immunoglobulin, neutrophil defensin, pyruvate kinase and alpha-enolase. The qualitative analysis considered only the exclusive proteins of each group. For the symptomatic group, 318 complete proteins and 29 fragments were identified, such as dedicator of cytokinesis protein, intersectin, prostaglandin, phospholipase DDHD2 and superoxide dismutase. For the asymptomatic group, 326 complete proteins and 37 fragments were identified, including azurocidin, C-reactive protein, collagen alpha, cathepsin, heat shock and laminin. CONCLUSIONS: Quantitative differences in the expression of common proteins in cases of symptomatic and asymptomatic AP were found, which were mostly related to host immune response in both groups. Exclusive proteins in the symptomatic group were mainly related to the host response to the presence of viruses in endodontic infections, oxidative stress and proteolytic enzymes. The results provide a basis for a better understanding of cellular and molecular pathways involved in AP, establishing specific proteomic profiles for symptomatic and asymptomatic conditions.

[Use of the Montreal Cognitive Assessment (MoCA) in Latin America: a systematic review]. / Uso del test de evaluacion cognitiva de Montreal (MoCA) en America Latina: revision sistematica.

AIM: To analyze the frequency of use of the Montreal Cognitive Assessment (MoCA) as a cognitive screening instrument, cross-cultural adaptation, the existence of normative data and clinical validation studies in Latin America Hispanic countries. SUBJECTS AND METHODS: The Medline, PsycINFO, Web of Science, Scopus and Scielo databases were consulted between April and June 2017 according to the PRISMA methodology. We included all studies referencing the MoCA as an instrument to evaluate cognitive deterioration conducted in Latin America and that included normative and psychometric data, as well as its clinical validation. RESULTS: Of the 80 studies identified, 19 met the inclusion criteria. Most of the studies mentioned the use of the Spanish version of the MoCA and reported a statistically significant effect of gender, age and, most of all, education on the performance of this test. Only five studies presented with a detailed analysis of the psychometric characteristics of the test, and in most articles cut-off scores for the diagnosis of cognitive impairment were the same as the original study. CONCLUSIONS: The small number of articles identified may reflect a late start of the use of MoCA in Latin America. A tendency towards the use of this test without making a cross-cultural adaptation and the use of international norms was observed in this region. The present systematic review demonstrates the need for future research tackling the development of a linguistically adapted version of the MoCA to Latin America and the study of its psychometric properties, with the aim of improving cognitive assessment.

TITLE: Uso del test de evaluacion cognitiva de Montreal (MoCA) en America Latina: revision sistematica.Objetivo. Analizar la frecuencia del uso del test de evaluacion cognitiva de Montreal (MoCA) como instrumento de cribado cognitivo, su adaptacion transcultural, la existencia de baremos y estudios de validacion clinica en paises de habla hispana en America Latina. Sujetos y metodos. Se ha realizado una revision sistematica de todos los estudios desarrollados en America Latina, con referencia al MoCA, que incluyan datos normativos, datos psicometricos y estudios de baremacion o de validacion clinica. Fueron consultadas sistematicamente, entre abril y junio de 2017, las bases de datos Medline, PsycINFO, Web of Science, Scopus y Scielo, conforme a la metodologia PRISMA. Resultados. De los 80 estudios encontrados, 19 cumplieron los criterios de inclusion. La mayoria de los estudios menciona el uso de la version española y comunica un efecto significativo del sexo, la edad y la escolaridad. El punto de corte adoptado por la mayoria de los autores para el diagnostico de deterioro cognitivo es el mismo del estudio original. Conclusiones. El reducido numero de articulos identificados refleja posiblemente un inicio tardio de la utilizacion del MoCA en America Latina. Esto pone de manifiesto una tendencia en la region a utilizar la prueba sin hacer una adaptacion transcultural de la version original y sin recurrir a normas internacionales para el diagnostico. La presente revision sistematica demuestra la necesidad de trabajos futuros de investigacion que puedan ofrecer una version linguisticamente adaptada del MoCA para America Latina y un estudio de sus propiedades psicometricas, con miras a una evaluacion cognitiva de mayor calidad.

Polineuropatia periférica na doença de Parkinson: prevalência e fatores de risco em uma amostra na cidade de Campina Grande - PB / Peripheral neuropathy in Parkinson's disease: prevalence and risk factors in a sample of Campina Grande city - Paraíba

Polineuropatia periférica (PNP) tem sido descrita na doença de Parkinson idiopática (DP), porém a prevalência e os fatores de risco não estão bem definidos. Objetivo: Investigar a prevalência e os fatores de risco para PNP na DP, em comparação com a população geral. Método: Participaram 36 pacientes com DP recrutados no ambulatório de Neurologia do Hospital Universitário Alcides Carneiro (HUAC) da Universidade Federal de Campina Grande (UFCG), Paraíba, e 30 sujeitos controles. Todos os participantes foram submetidos a caracterização clínica da PNP, ao estudo de neurocondução (ENC) dos nervos peroneal e sural bilateral e as dosagens de vitamina B12, homocisteina, ácido metilmalônico e ácido fólico. A Escala Unificada de Avaliação da Doença de Parkinson - III e a de Hoehn-Yahr foram utilizadas na avaliação motora do grupo Parkinson (GP). Resultados: Sinais e sintomas neuropáticos foram mais frequentes no GP (61%). Alterações nos parâmetros do ENC foram observadas em 44,4% do GP e 26,7% do grupo controle, sendo a PNP confirmada em três pacientes e um controle. Análise de regressão revelou associação significativa entre os sintomas neuropáticos e a DP, sem associação com aspectos clínicos e bioquímicos. Conclusão: Pacientes com DP possuem maiores escores neuropáticos e maior prevalência de PNP que controles. Os dados sugerem a própria DP como fator de risco para o desenvolvimento da PNP, minimizando o papel da vitamina B12 e de seus metabólitos neste processo.(AU)

Peripheral neuropathy (PN) has been described in idiopathic Parkinson disease (PD) however the prevalence and the risk factors are not well established. Objective: To assess the prevalence of PN and the risk factors for neuropathy in PD against the general population. Method: Participated in the study 36 PD patients recruited from Neurology Outpatient Unit of Hospital Universitário Alcides Carneiro of the Federal University of Campina Grande, Paraíba, and 30 controls. All the participants were submitted to clinical characterization of PN, nerve conduction study (NCS) and biochemical dosages (B12 vitamin, homocysteine, methylmalonic acid and folic acid). Results: Neuropathic signs and symptoms were more frequent in PD (61%). Alterations in parameters of NCS were observed in 44.4% of Parkinson group and 26.7% of control group, and PN was confirmed in 3 PD patients and 1 control. Regression analyses showed a significant association between symptoms of PN and PD, without association with clinical and biochemical features. Conclusion: PD patients have higher neuropathic scores and frequency of PN than controls. Data suggests the PD by itself as a risk factor for development of PN, reducing the role of B12 vitamin and its metabolites in this process.(AU)

A ten-year experience and follow-up of three hundred patients fitted with the Biomet/Lorenz Microfixation TMJ replacement system.

The purpose of this paper is to present the postoperative results obtained after full temporomandibular joint (TMJ) reconstruction employing the Biomet/Lorenz Microfixation TMJ replacement system (Jacksonville, FL, USA) in 300 patients (201 unilateral, 99 bilateral). Objective data (maximum inter-incisal opening; MIO) and subjective data (function and speech, diet, and pain) were collected preoperatively and at postoperative evaluations performed over a 10-year period (mean 3.5, standard deviation 2.1 years). The MIO measures were obtained using a calliper rule. Subjective data were evaluated using a visual analogue scale with scores ranging from 0 to 5 for each variable. The results were analyzed with the paired t-test (two-sided, α=5%). Each patient showed significant improvements for all of the variables at evaluation on postoperative day 7. The results for MIO, function and speech, and diet, showed improvements at each postoperative evaluation over a maximum of 3 years, with stabilization of the results from the fourth year. Complaints of pain decreased considerably up to the 1-month postoperative evaluation, and no patient reported severe pain at 6 months after surgery. The results presented show that the reconstruction of the TMJ through the installation of the Biomet/Lorenz system prosthesis is a safe and effective option for proper reestablishment of the joint and stomatognathic system function; significant long-term improvements in mandibular range of motion are promoted and pain levels decrease.

Treatment for agenesis of maxillary lateral incisors: a systematic review.

OBJECTIVES: To compare the efficacy and safety of three orthodontic treatment modalities for agenesis of maxillary lateral incisors: 1) closing the space with the reshaped canine substituting the lateral incisor, 2) opening the space with placement of a conventional fixed bridge, and 3) opening the space with placement of a single-unit implant and an implant-supported crown. SETTING: Brazilian Cochrane Center and Universidade Federal de São Paulo, Brazil. MATERIAL AND METHODS: The following databases were investigated: Cochrane Register of Controlled Trials (Edition 12, 2011), EMBASE (from 1974 to December 2011), MEDLINE (from 1965 to December 2011), LILACS (from 1966 to November 2011), and Odontology Brazilian Bibliography Database (from 1966 to November 2011). Conference abstracts, main Brazilian dissertations and theses databases, and reference lists were handsearched. This systematic review included randomized or quasi-randomized controlled trials (RCTs) including women aged 15 years or over and men aged 21 years or over who received one of the interventions stated above. Two observers independently evaluated all the studies regarding eligibility criteria and assessed the risk of bias of included studies. RESULTS: No studies were included in the review as no RCTs were found. Most of the evidence comes from case reports and narrative reviews on case reports and from three studies with a single post-intervention evaluation and non-comparable control groups with high risk of bias. CONCLUSIONS: There is no scientific evidence for any of the three most common types of treatment for agenesis of the maxillary lateral incisors. RCTs into this issue are still necessary.

Rendimentos de carcaça, composição tecidual e musculosidade da perna de cordeiros alimentados com silagem de milho ou cana-de-açúcar em dois níveis de concentrado / Carcass yield, tissue composition, and leg muscularity of lambs fed corn silage or sugar cane in two levels of concentrate

Avaliou-se o efeito da relação volumoso:concentrado e do tipo de volumoso nos rendimentos de carcaça e dos cortes comerciais, e sobre a composição dos tecidos e musculosidade da perna de 32 cordeiros Ile de France, terminados em confinamento, alimentados com silagem de milho ou cana-de-açúcar, em duas relações volumoso:concentrado: de 60:40 ou 40:60. Os cordeiros foram abatidos aos 32kg de peso corporal; calcularam-se o peso e o rendimento da carcaça quente (PCQ e RCQ). Após 24 horas de refrigeração, obtiveram-se o peso e o rendimento da carcaça fria (PCF e RCF), o rendimento verdadeiro e as perdas por resfriamento. As carcaças foram divididas em cinco cortes comerciais, paleta, lombo, perna, costelas e pescoço, e a perna foi dissecada em músculo, osso e gordura, calculando-se as relações músculo:osso e músculo:gordura, além do índice de musculosidade da perna. Os cordeiros alimentados com silagem de milho apresentaram maiores PCQ (16,05kg), PCF (15,61kg), RCQ (50,4 por cento) e RCF (49,0 por cento). O rendimento de lombo foi menor nos cordeiros alimentados com cana-de-açúcar (9,1 por cento). A cana-de-açúcar e a silagem de milho nas duas relações volumoso:concentrado estudadas não afetaram as proporções de osso, músculo e gordura e musculosidade da perna, e podem constituir a dieta desses animais sem diminuir as proporções desses tecidos, responsáveis pela qualidade da carne.

The effects of roughage:concentrate ratio and type of roughage were evaluated on the carcass yield and commercial cuts, and tissue composition and leg muscularity of 32 Ile de France lambs finished in feedlot and fed corn silage or sugar cane in two roughage:concentrate ratios: 60:40 or 40:60. The lambs were slaughtered with 32kg of body weight and the weight and yield of hot carcass (WHC and YHC) were determined. After refrigerated for 24h in cold camera, the weight and yield of cold carcass (WCC and YCC), real yield, and weight loss for cooling were obtained. The carcass was divided in five commercial cuts (shoulder, loin, leg, ribs, and neck), and the leg was dissected on muscle, bone, and fat, and the relations muscle:bone and muscle:fat and also the leg muscularity index were calculated. The lambs fed corn silage showed higher WHC (16.05kg), WCC (15.61kg), YHC (50.42 percent), and YCC (49.03 percent). The loin yield was lower in lambs fed sugar cane (9.11 percent). The sugar cane and corn silage in this two roughage:concentrate ratios did not affect the proportions of muscle, bone, and fat, and leg muscularity. Therefore, they can be used in diets for animals without reducing the proportions of these tissues, that are related to meat quality.

Rendimentos de carcaça, composição tecidual e musculosidade da perna de cordeiros alimentados com silagem de milho ou cana-de-açúcar em dois níveis de concentrado / Carcass yield, tissue composition, and leg muscularity of lambs fed corn silage or sugar cane in two levels of concentrate

Avaliou-se o efeito da relação volumoso:concentrado e do tipo de volumoso nos rendimentos de carcaça e dos cortes comerciais, e sobre a composição dos tecidos e musculosidade da perna de 32 cordeiros Ile de France, terminados em confinamento, alimentados com silagem de milho ou cana-de-açúcar, em duas relações volumoso:concentrado: de 60:40 ou 40:60. Os cordeiros foram abatidos aos 32kg de peso corporal; calcularam-se o peso e o rendimento da carcaça quente (PCQ e RCQ). Após 24 horas de refrigeração, obtiveram-se o peso e o rendimento da carcaça fria (PCF e RCF), o rendimento verdadeiro e as perdas por resfriamento. As carcaças foram divididas em cinco cortes comerciais, paleta, lombo, perna, costelas e pescoço, e a perna foi dissecada em músculo, osso e gordura, calculando-se as relações músculo:osso e músculo:gordura, além do índice de musculosidade da perna. Os cordeiros alimentados com silagem de milho apresentaram maiores PCQ (16,05kg), PCF (15,61kg), RCQ (50,4 por cento) e RCF (49,0 por cento). O rendimento de lombo foi menor nos cordeiros alimentados com cana-de-açúcar (9,1 por cento). A cana-de-açúcar e a silagem de milho nas duas relações volumoso:concentrado estudadas não afetaram as proporções de osso, músculo e gordura e musculosidade da perna, e podem constituir a dieta desses animais sem diminuir as proporções desses tecidos, responsáveis pela qualidade da carne.(AU)

The effects of roughage:concentrate ratio and type of roughage were evaluated on the carcass yield and commercial cuts, and tissue composition and leg muscularity of 32 Ile de France lambs finished in feedlot and fed corn silage or sugar cane in two roughage:concentrate ratios: 60:40 or 40:60. The lambs were slaughtered with 32kg of body weight and the weight and yield of hot carcass (WHC and YHC) were determined. After refrigerated for 24h in cold camera, the weight and yield of cold carcass (WCC and YCC), real yield, and weight loss for cooling were obtained. The carcass was divided in five commercial cuts (shoulder, loin, leg, ribs, and neck), and the leg was dissected on muscle, bone, and fat, and the relations muscle:bone and muscle:fat and also the leg muscularity index were calculated. The lambs fed corn silage showed higher WHC (16.05kg), WCC (15.61kg), YHC (50.42 percent), and YCC (49.03 percent). The loin yield was lower in lambs fed sugar cane (9.11 percent). The sugar cane and corn silage in this two roughage:concentrate ratios did not affect the proportions of muscle, bone, and fat, and leg muscularity. Therefore, they can be used in diets for animals without reducing the proportions of these tissues, that are related to meat quality.(AU)

Subgenotype diversity of hepatitis B virus American genotype F in Amerindians from Venezuela and the general population of Colombia.

The objective of this study was the evaluation of the genetic diversity found in HBV circulating among Venezuelan Amerindians and the general population in Colombia. Phylogenetic analysis of the S region in 194 isolates showed that genotype F is highly predominant in Colombia and Venezuela. This might be related to the genetic background of the population. F3 is the main subgenotype which circulates in both countries. Phylogenetic analysis of 61 complete genome sequences of HBV American genotypes confirms the presence of two genotypes F and H, and 4 F subgenotypes. In Venezuela, subgenotypes F1, F2, and F3 circulate in East and West Amerindians, while only F3 was found among South Amerindians. Japreira community derived from Yucpa Amerindians around 150 years ago. However, several Japreira HBV sequences were forming a clade that can be classified as subgenotype 2b, differing from Yucpa sequences that belong mainly to subgenotype F3. The apparent absence of correlation between the phylogenetic groupings of HBV isolates with the ethnical origin in aboriginal populations might be suggesting a recent origin of HBV American subgenotypes, or a genetic drift effect.

Body fat in men with prolactinoma.

OBJECTIVE: (a) To evaluate body fat in men with prolactinoma and healthy controls, using whole body dual energy x-ray absorptiometry (DXA), and (b) to correlate DXA results with anthropometry and clinical aspects of male prolactinomas. MATERIAL AND METHODS: A cross-sectional study was performed in two University referral centers. Eleven newly-diagnosed men with prolactinoma and 9 with normal PRL levels due to dopamine agonist treatment were submitted to DXA and blood analysis (PRL, testosterone, dihydrotestosterone, estradiol, and SHBG) by the time of their clinical evaluation. They were compared with 14 control men of similar age and body mass index distribution. RESULTS: Newly-diagnosed men with prolactinoma had higher fat percentage in the arms and the total body, when compared with patients treated with dopamine agonists and controls. The former group also presented higher fat percentage in the legs than the controls. Truncal fat percentage of the newly-diagnosed patients was lower than the dopamine agonist treated group. The 3 groups had similar android and gynoid fat contents. Fat percentage of the 6 sites correlated with PRL, testosterone, and dihydrotestosterone levels. CONCLUSION: Newly-diagnosed men with prolactinomas had higher body fat content. Body fat was linked to disease control, especially to the PRL and androgen levels. Consequently, adequate control of hyperprolactinemia should be pursued in order to reduce the risk of obesity and its metabolic complications in men with prolactinoma.

Sensible and latent heat loss from the body surface of Holstein cows in a tropical environment.

The general principles of the mechanisms of heat transfer are well known, but knowledge of the transition between evaporative and non-evaporative heat loss by Holstein cows in field conditions must be improved, especially for low-latitude environments. With this aim 15 Holstein cows managed in open pasture were observed in a tropical region. The latent heat loss from the body surface of the animals was measured by means of a ventilated capsule, while convective heat transfer was estimated by the theory of convection from a horizontal cylinder and by the long-wave radiation exchange based on the Stefan-Boltzmann law. When the air temperature was between 10 and 36 degrees C the sensible heat transfer varied from 160 to -30 W m(-2), while the latent heat loss by cutaneous evaporation increased from 30 to 350 W m(-2). Heat loss by cutaneous evaporation accounted for 20-30% of the total heat loss when air temperatures ranged from 10 to 20 degrees C. At air temperatures >30 degrees C cutaneous evaporation becomes the main avenue of heat loss, accounting for approximately 85% of the total heat loss, while the rest is lost by respiratory evaporation.

Infection with transfusion-transmitted virus (TTV) in humans and other primates in Venezuela.

Tranfusion-transmitted virus (TTV), a single-stranded circular DNA virus that chronically infects humans and other animals, displays a high degree of genetic diversity and was originally thought to be associated with hepatitis. The prevalences of TTV infection among different populations of humans and non-human primates from Venezuela have now been evaluated, using serum samples and three different detection tests. All three tests were PCR-based, one involving a hemi-nested PCR and primers based on the N22 open-reading-frame-1 region (N22-PCR), another employing 55 cycles with primers from the more conserved untranslated region (UTR-PCR), and the other using a hemi-nested PCR with primers from the same region (HUTR-PCR). The overall prevalences of human infection appeared much higher with the HUTR-PCR (52%) than with the N22-PCR (13%) or the UTR-PCR (5%). When the products amplified by N22-PCR from 28 human isolates of TTV were sequenced, only two genotypes of the virus were detected. The non-human sera tested came from primates kept in a zoo in north-western Venezuela. TTV DNA was detected, by HUTR-PCR, in both of the chimpanzee sera tested but not in any of the sera from the 11 New-World primates or the other 12 Old-World primates that were investigated. The results, particularly those of the HUTR-PCR, indicate that TTV infection is common in Venezuela, especially in populations, such as many Amerindian groups, who live under poor sanitary conditions. Although TTV infection may be relatively rare among non-human primates from the New World, this will have to be investigated further, using many more samples collected throughout the Americas.

Hepatitis delta virus genotypes I and III circulate associated with hepatitis B virus genotype F In Venezuela.

The genotypes of hepatitis B (HBV) and delta (HDV) viruses circulating among Venezuelan Amerindian populations, where these viruses are endemic, were determined by sequencing of PCR amplified products from HBsAg positive sera. HDV genotype I (n = 7, 6 from West Amerindians), and III (n = 5, 4 from South Amerindians), were found. Only one HDV genotype I isolate was associated with HBV genotype D, 4 HDV genotype I and 2 HDV genotype III infected individuals were co-infected with HBV genotype F. The failure to detect the South American HDV genotype III in West Amerindians might be related to the outbreak of fulminant hepatitis with high mortality rate occurred between 1979 and 1982, probably affecting more the Amerindians infected with HDV genotype III. These results suggest the circulation of HDV genotype I among Amerindians, probably introduced through European immigrations, and that this HDV genotype is able to replicate in association with HBV genotype F.

Prevalence of infection with hepatitis C virus in Venezuela, as assessed with an immuno-assay based on synthetic peptides.

Information on infection with hepatitis C virus (HCV) in South America is scarce. The seroprevalences of antibodies to HCV among urban, rural and Amerindian populations from Venezuela, and the genotypes of the HCV isolates recovered, were therefore determined. A total of 2592 sera were tested with an immuno-assay which was developed in-house and based on synthetic peptides. Each reactive sample was then re-tested, using other enzyme immuno-assays and a reverse-transcription, nested PCR, and any sample confirmed positive (in any test) was considered HCV-positive. Genotypes were determined by analysis of RFLP. Overall, 39 (1.5%) of the samples were found HCV positive. The results of the immuno-assays indicated that the seroprevalence of HCV markers among the Amerindians investigated (23/1082, or 2.1%) was significantly higher than that among the other subjects (16/1510, or 1.1%; P = 0.02). No such difference was observed in the numbers of subjects confirmed positive by PCR, however (6/1082 v. 10/1510), and some of the anti-HCV reactivity observed among Amerindians may have been the result of cross-reactivity with parasitic infections. The relative low prevalence of active HCV infection (16/2582, or 0.6%) and the HCV genotypes observed (mainly genotype 1) are in agreement with the results of previous studies indicating that HCV is not autochthonous to South America. However, it is clear that the virus may now be found even in isolated Amerindian populations. The in-house, synthetic-peptide-based immuno-assay seems to be a valuable tool for epidemiological studies.

Hepatitis B virus DNA in blood samples positive for antibodies to core antigen and negative for surface antigen.

Anti-hepatitis B core antigen (HBcAg)-positive hepatitis B surface antigen (HBsAg)-negative plasma samples from blood donors were tested by nested PCR. DNA positivity was more significantly associated with high levels of anti-HBcAg than with low levels of anti-HBsAg antibodies. Analysis of a dilution of anti-HBcAg antibodies might result in a more rational exclusion of anti-HBcAg-positive HBsAg-negative samples, reducing the number of donations discarded and enabling more countries to incorporate anti-HBcAg testing.

Turnover of hepatitis C virus genotypes in hemodialysis patients.

Hepatitis C virus (HCV) genotypes were determined in hemodialysis patients with a high prevalence and incidence of infection. A change of HCV genotype was observed in 6/14 follow-up samples analyzed 13 and 21 months later. The appearance and disappearance of HCV genotypes may be due to either genotype-specific intermittent viremic status or viral interference.

Norwalk virus infection in Venezuela.

The presence of antibodies against Norwalk virus (NV) was studied in sera from different Venezuelan populations, using an enzyme immuno-assay (EIA) based on recombinant NV protein. Antibodies to NV were found in 47%-53% of urban subjects from Caracas, 83% of rural subjects from the west of the country, and 73%-93% of Amerindian subjects. The prevalences found in the rural and Amerindian groups were significantly higher than that in the urban group. Although about 50% of the children studied were seropositive for NV by the age of 5 years, only four (0.4%) of 1120 faecal samples from children with diarrhoea which were tested for the presence of NV antigen by sandwich EIA were found positive. An increase of at least 4-fold in the titre of anti-NV IgA was found in three (5%) of 61 pairs of sera taken during and 1 month after an acute episode of diarrhoea not due to rotavirus. NV was therefore not a predominant aetiological cause of gastro-enteritis in young children in Venezuela between 1993 and 1995, although it can be the cause of diarrhoea in infants.

Hepatitis G virus infection in Amerindians and other Venezuelan high-risk groups.

Recently, a new virus related to flaviviruses, the hepatitis G virus (HGV), or GBV-C virus, was discovered as a putative blood-borne human pathogen. HGV RNA (NS5 region) was amplified by reverse transcription-nested PCR in the sera of 6 of 64 (9%) hemodialysis patients; 2 of 80 (2.5%) West Yukpa Amerindians, a population with a high rate of HBV infection but negative for HCV infection; and 1 patient with an acute episode of non-A, non-B, non-C hepatitis (NABCH). The patterns of single-strand conformation polymorphism of the amplified products were unique among different specimens and similar on follow-up for hemodialysis patients. All patients tested remained HGV RNA positive 1 and 2 years later, without major sequence variation, except for the NABCH patient, for whom a double infection and an apparent clearance of the original dominant variant was observed after 2 years. The sequences of the NS5 amplified products demonstrated 85 to 90% identity with other reported HGV sequences.

Determination of genotypes of hepatitis C virus in Venezuela by restriction fragment length polymorphism.

Hepatitis C virus genotypes in Venezuela were analyzed by restriction fragment length polymorphism in the 5' noncoding region. The absence of BstUI digestion was found to be a useful marker for genotype 2 specimens. From 122 serum samples, 66, 20, and 2.5% were classified as genotypes 1, 2, and 3, respectively; 0.8% were classified as genotype 4; and 10% appeared to be mixed infections.