Selected ruminal variables and purine urinary excretion rate of steers subjected to feeding, fasting, and re-feeding conditions.

The effects of feeding, fasting, and re-feeding on the ruminal profile of growing cattle were studied. Ruminal fluid and urine samples were obtained from 12 crossbred steers weighing approximately 300 kg during the following periods: 11 h of normal feeding (postprandial period), 48 consecutive hours of fasting, and followed by 48 h of re-feeding. Fasting promotes changes in the ruminal profile, such as an increase in ruminal pH, reduction in the number of rumen protozoa and bacteria, and decrease in the urinary excretion of allantoin; however, it does not change the urinary uric acid excretion rate. The overall mean ruminal pH was higher during fasting (7.53±0.27) in comparison to those at normal feeding (6.72±0.25) and re-feeding (6.62±0.31) (p⟨0.05). During re-feeding, the ruminal profile returned to normal, except for the protozoa count, which despite a slight increase only after 48 h of re-feeding, did not recover to baseline values.

Detection of East/Central/South African genotype Chikungunya virus during an outbreak in a southeastern state of Brazil.

Chikungunya virus (CHIKV) is an arbovirus transmitted by Aedes mosquitoes that was first identified in Brazil in 2014. It causes a febrile illness characterised by severe arthralgia and rash. Our group investigated a suspected CHIKV outbreak in Governador Valadares, state of Minas Gerais, Brazil and from 25 acute-phase patients, 10 had qRT-PCR positive sera samples and had E1 partial sequence amplified and Sanger sequenced. Samples were identified as East/Central/South African (ECSA) genotype by phylogenetic analysis and clustered with CHIKV sequences isolated in the neighbour state of Bahia. Our findings confirm previous predictions that ECSA genotype would spread through northeast and southeast of Brazil.

Foreign body-induced changes in the reticular contraction pattern of sheep observed with M-mode ultrasonography.

In the pre-experimental period of a clinical trial, an apparently clinically healthy sheep fitted with ruminal and abomasal cannulas showed changes in the reticular contraction pattern visualized in M-mode ultrasonogram. Radiographic examination revealed a blunt metal screw in its reticulum. By the time change in the reticular motility through the ultrasound examination was detected, the animal had still not expressed any behavioral changes. A description of the clinical case, follow-up of the findings and laboratory data, like white blood cell count, serum pepsinogen and fibrinogen concentrations, were presented. The foreign body was removed through the ruminal cannula and reticular contraction tended to normal. An association of the contraction pattern with measured clinical data was possible, leading to the conclusion that use of M-mode ultrasonography has a potential application in similar clinical situations.

High rates of human immunodeficiency virus type 1 mutational profiles by single-genome amplification after 48-hour propagation in peripheral blood mononuclear cells at different levels of cell activation.

Human immunodeficiency virus type 1 (HIV-1) genetic diversity is one of the most important features of HIV-1 infections and the result of error accumulation during reverse transcription and of high viral turnover. HIV-1 reverse transcription is influenced by factors such as the level of nucleotides and/or the cellular activation state. HIV-1 diversity was investigated after 48 h of viral propagation in peripheral blood mononuclear cells (PBMCs) obtained from healthy donors in three different cell culture conditions: (1) resting PBMCs, (2) simultaneous infection and PBMC activation, and (3) PBMC activation 72 h before infection. Cellular DNA was extracted and proviruses of each culture condition were amplified. Single-genome PCR clones were obtained and the protease and reverse transcriptase of the pol gene were sequenced. An elevated number of nucleotide substitutions in all three culture conditions were observed. In condition 1, the mutational rate observed ranged from 1.0 × 10(-3) to 2.1 × 10(-2), the genetic diversity was 0.6%, and hypermutation was observed in 7.1% of sequenced clones. In condition 2, the mutational rate ranged from 1.0 × 10(-3) to 1.0 × 10(-2), the genetic diversity was 0.8%, and hypermutation affected 6.7% of clones. In condition 3, the mutational rate ranged from 2.8 × 10(-3) to 1.1 × 10(-2), the genetic diversity was 1%, and 5.9% of clones were hypermutated. Substitutions occurred more frequently in some specific nucleotide stretches, and a common pattern for substitutions in all the different conditions was identified. There was a significant accumulation of mutations during the initial periods of in vitro HIV-1 propagation irrespective of culture conditions. The rapid accumulation of virus diversity might represent a viral strategy when colonizing new hosts. Complementary studies are necessary to allow for a better understanding of the initial periods of infection, which represent a crucial event related to disease progression.

Avaliação de alguns tratamentos na intoxicação por amônia em bovinos / Evaluation of some treatments for ammonia poisoning in cattle

Para avaliar tratamentos na intoxicação por amônia, 15 novilhos, infundidos com solução de cloreto de amônio, foram distribuídos em três grupos de cinco animais cada e tratados, como: 1 - grupo-controle (C), infundido com solução salina fisiológica (H); 2 - grupo O+H, medicado com aminoácidos do ciclo da ureia (O) e "H"; 3 - grupo O+F+H, o mesmo protocolo do grupo 2 acrescido de furosemida (F). Os animais foram monitorados, colhendo-se amostras sanguíneas e todo volume urinário. Os tratamentos O+F+H e O+H promoveram melhora clínica pronunciada, em relação ao grupo H, isto é, permaneceram em estação, retornaram o movimento de rúmen e o apetite e recuperaram-se do edema pulmonar mais rapidamente. Observaram-se, nos grupos O+F+H e O+H, teores plasmáticos mais baixos de amônia e lactato-L, urina mais abundante, excreção pela urina de maior quantidade de amônio e ureia, depuração mais intensa de amônia do sangue e pH urinário mais baixo. Concluiu-se que os tratamentos propostos apresentaram ação efetiva principalmente por diminuírem a hiperamonemia, por meio da maior excreção renal de amônio e maior transformação hepática de amônia em ureia.

Fifteen yearling steers were used to evaluate treatments for ammonia intoxication. The animals subjected to ammonium chloride infusion were distributed into three groups of five animals each, and treated as following: 1 - control group (C), physiological saline solution (H) infusion; 2 - O+H group, urea cycle aminoacids (O) and "H" medication; and 3 - O+F+H group, the same protocol of group 2 plus furosemide (F). The animals were monitored by collection of blood samples and the whole urinary volume. O+F+H and O+H treatments promoted pronounced clinical improvement, in attention to H group, i.e., they remained in standing position, returned ruminal movement and appetite, and recovered more quickly from pulmonary edema. In O+F+H and O+H groups, minor plasmatic levels of ammonia and L-lactate, more abundant urine, major ammonium and urea amount excretion by urine, more intensive ammonia depuration from blood, and lower urinary pH were observed. It was concluded that the proposed treatments had effective action due to hyperammonemia decrease, by major renal ammonium excretion, and hepatic transformation of ammonia in urea.

Avaliação de alguns tratamentos na intoxicação por amônia em bovinos / Evaluation of some treatments for ammonia poisoning in cattle

Para avaliar tratamentos na intoxicação por amônia, 15 novilhos, infundidos com solução de cloreto de amônio, foram distribuídos em três grupos de cinco animais cada e tratados, como: 1 - grupo-controle (C), infundido com solução salina fisiológica (H); 2 - grupo O+H, medicado com aminoácidos do ciclo da ureia (O) e "H"; 3 - grupo O+F+H, o mesmo protocolo do grupo 2 acrescido de furosemida (F). Os animais foram monitorados, colhendo-se amostras sanguíneas e todo volume urinário. Os tratamentos O+F+H e O+H promoveram melhora clínica pronunciada, em relação ao grupo H, isto é, permaneceram em estação, retornaram o movimento de rúmen e o apetite e recuperaram-se do edema pulmonar mais rapidamente. Observaram-se, nos grupos O+F+H e O+H, teores plasmáticos mais baixos de amônia e lactato-L, urina mais abundante, excreção pela urina de maior quantidade de amônio e ureia, depuração mais intensa de amônia do sangue e pH urinário mais baixo. Concluiu-se que os tratamentos propostos apresentaram ação efetiva principalmente por diminuírem a hiperamonemia, por meio da maior excreção renal de amônio e maior transformação hepática de amônia em ureia.(AU)

Fifteen yearling steers were used to evaluate treatments for ammonia intoxication. The animals subjected to ammonium chloride infusion were distributed into three groups of five animals each, and treated as following: 1 - control group (C), physiological saline solution (H) infusion; 2 - O+H group, urea cycle aminoacids (O) and "H" medication; and 3 - O+F+H group, the same protocol of group 2 plus furosemide (F). The animals were monitored by collection of blood samples and the whole urinary volume. O+F+H and O+H treatments promoted pronounced clinical improvement, in attention to H group, i.e., they remained in standing position, returned ruminal movement and appetite, and recovered more quickly from pulmonary edema. In O+F+H and O+H groups, minor plasmatic levels of ammonia and L-lactate, more abundant urine, major ammonium and urea amount excretion by urine, more intensive ammonia depuration from blood, and lower urinary pH were observed. It was concluded that the proposed treatments had effective action due to hyperammonemia decrease, by major renal ammonium excretion, and hepatic transformation of ammonia in urea.(AU)

Comparative effectiveness of dried plasma HIV-1 viral load testing in Brazil using ViveST for sample collection.

BACKGROUND: Utilization of dried plasma for HIV-1 viral load testing would significantly decrease sample shipping costs. OBJECTIVES: To describe the precision and reproducibility of ViveST(®) (ST) as a transportation method for shipping specimens for HIV-1 viral load (VL) testing. STUDY DESIGN: Thirty clinical plasma samples were used to generate replicate samples with HIV VL values of 4 log(10), 3 log(10) and 2 log(10) copies/mL for reproducibility testing and an additional 299 samples with HIV VL <50 copies/mL (99); 1.7 log(10) to 3.99 log(10) (100); and 4 log(10) to 5.99 log(10)/mL (100) were used to compare ViveST to frozen plasma samples using the VERSANT(®) HIV-1 RNA 3.0 Assay. Results were compared using Student t-test, Pearson correlation and Bland-Altman analyses. RESULTS: Mean intra-assay variance among frozen and dried plasma triplicates was 0.15 log(10) and 0.09 log(10) copies/mL respectively (n=10, P=NS). Compared to frozen plasma, there was a mean reduction of 0.3 log(10), 0.27 log(10), and 0.35 log(10) copies/mL at the 4 log(10), 3 log(10), and 2 log(10) copy/mL samples respectively (n=30, all comparisons, P<0.01). Overall correlation between 299 frozen and ViveST samples was r=0.97, where 12 of 99 undetectable frozen VL were positive with ST, and 12 of 200 frozen detectable VL were undetectable with ViveST (mean VL 2.1, 1.9 log(10) copies/mL respectively). CONCLUSIONS: HIV-1 viral load results using ViveST were reproducible, correlated well with frozen plasma, though yielding minimally lower values. Our data suggest that dried plasma for HIV-1 VL testing using ViveST has promise for use in HIV clinical practice.

Prevalence, distribution and correlates of endocervical human papillomavirus types in Brazilian women.

We determined the prevalence, distribution and correlates of human papillomavirus (HPV) types in 386 mixed-income, sexually active women in São Paulo, Brazil. Endocervical samples were tested for HPV DNA with L1 primers MY09 and MY11; negative and indeterminate samples were retested using GP 5+/6+ consensus primers. HPV was detected in 35% of all women; high-risk/probable high-risk types in 20%; low-risk types in 7%; and an indeterminate type in 10%. Twenty-five HPV types were found overall: 17 (probable) high-risk types and eight low-risk types. Approximately one-third (29%) of women with HPV infection were positive for type 16 or 18 and 36% were positive for types 6, 11, 16 or 18. The presence of (probable) high-risk HPV was associated with younger age, more lifetime sex partners and abnormal vaginal flora. Additional studies mapping the distribution of HPV types worldwide are necessary to prepare for vaccination programmes and direct future vaccine development.

Effects of gamma radiation on the growth of Alternaria alternata and on the production of alternariol and alternariol monomethyl ether in sunflower seeds.

The objective of the present study was to evaluate the effects of different gamma radiation doses on the growth of Alternaria alternata and on the production of toxins alternariol (AOH), and alternariol monomethyl ether (AME) in sunflower seed samples. After irradiation with 2, 5 and 7 kGy, the spore mass was resuspended in sterile distilled water and the suspension was inoculated into sunflower seeds. The number of colony-forming units per gram (CFU/g) was determined after culture on Dichloran Rose Bengal Chloramphenicol and Dichloran Chloramphenicol Malt Extract Agar. The presence of AOH and AME was investigated by liquid chromatography coupled to mass spectrometry. The radiation doses used resulted in a reduction of the number of A. alternata CFU/g and of AOH and AME levels when compared to the nonirradiated control group. Maximum reduction of the fungus (98.5%) and toxins (99.9%) was observed at a dose of 7 and 5 kGy, respectively. Under the present conditions, gamma radiation was found to be an alternative for the control of A. alternata and, consequently, of AOH and AME production in sunflower seeds.

Effect of an energy-deficient diet on populations of ciliate protozoans in bovine rumen / Efeito da deficiência de energia na dieta sobre a população de protozoários ciliados do rúmen de bovinos

Ten young rumen-cannulated crossbred steers were randomly divided into two groups: a control group (C; n=4), which was fed a balanced diet for daily weight gain of 900g; and a pronounced energy-deprived group (PED; n=6), receiving 30 percent less of the required energy for maintenance. After 140 days of these alimentary regimes, rumen fluid and urine samples were collected for biochemical and functional tests, before feeding and at 1, 3, 6, and 9 hours after feeding. The energy-deprivation diet caused a significant reduction in the number of Entodinium, Eodinium, Isotricha, Dasytricha, Eremoplastron, Eudiplodinium, Metadinium, Charonina, Ostracodinium, and Epidinium protozoa. There was no effect of the time of sampling in both groups on the total number of ciliates in rumen fluid. A higher number of protozoan forms in binary division were recorded in the control group, at the 6th and 9th hours after feeding (P<0.019). There was a high positive correlation between the total count of protozoans in rumen fluid and glucose fermentation, ammonia, and urinary allantoin excretion index; and a negative correlation between the total count of protozoa and metilene blue reduction, and a medium correlation between the total count of protozoa and total volatile fatty acids concentration. The determination of the protozoa populations does not imply in the use of complex and hard-to-execute techniques, although it is time consuming and needs practice. This exam particularly helps in clinical expected diagnosis.

Foram utilizados 10 novilhos mestiços com cânula ruminal, distribuídos em dois grupos: no grupo controle (C; n=4) receberam dieta balanceada para ganho diário de 900g; no grupo tratado com carência pronunciada de energia (CP; n=6), receberam dieta com 30 por cento a menos do nível de mantença em energia. Após 140 dias sob esses regimes de alimentação, foram coletadas amostras do fluido ruminal e urina, para realização de provas bioquímicas e funcionais, antes e às 1, 3, 6 e 9 horas após o fornecimento do alimento. A carência energética resultou em diminuição significativa na quantidade dos protozoários Entodinium, Eodinium, Isotricha, Dasytricha, Eremoplastron, Eudiplodinium, Metadinium, Charonina, Ostracodinium e Epidinium. Não houve efeito da hora de coleta sobre o total de ciliados nos grupos C e CP. Maior número de formas em divisão binária foi registrado no grupo C, na sexta e nona horas pós-alimentação (P<0,019). Observaram-se altas correlações positivas entre a contagem total de protozoários e a fermentação de glicose, amônia e o índice de excreção urinária de alantoína e negativa entre a contagem total de protozoários e a redução do azul de metileno, e correlação média entre a contagem total de protozoários e os ácidos graxos voláteis totais. A determinação da população de protozoários do rúmen é um método simples de avaliação, além de que particularmente auxilia o diagnóstico clínico da função ruminal.

Effect of an energy-deficient diet on populations of ciliate protozoans in bovine rumen / Efeito da deficiência de energia na dieta sobre a população de protozoários ciliados do rúmen de bovinos

Ten young rumen-cannulated crossbred steers were randomly divided into two groups: a control group (C; n=4), which was fed a balanced diet for daily weight gain of 900g; and a pronounced energy-deprived group (PED; n=6), receiving 30 percent less of the required energy for maintenance. After 140 days of these alimentary regimes, rumen fluid and urine samples were collected for biochemical and functional tests, before feeding and at 1, 3, 6, and 9 hours after feeding. The energy-deprivation diet caused a significant reduction in the number of Entodinium, Eodinium, Isotricha, Dasytricha, Eremoplastron, Eudiplodinium, Metadinium, Charonina, Ostracodinium, and Epidinium protozoa. There was no effect of the time of sampling in both groups on the total number of ciliates in rumen fluid. A higher number of protozoan forms in binary division were recorded in the control group, at the 6th and 9th hours after feeding (P<0.019). There was a high positive correlation between the total count of protozoans in rumen fluid and glucose fermentation, ammonia, and urinary allantoin excretion index; and a negative correlation between the total count of protozoa and metilene blue reduction, and a medium correlation between the total count of protozoa and total volatile fatty acids concentration. The determination of the protozoa populations does not imply in the use of complex and hard-to-execute techniques, although it is time consuming and needs practice. This exam particularly helps in clinical expected diagnosis.(AU)

Foram utilizados 10 novilhos mestiços com cânula ruminal, distribuídos em dois grupos: no grupo controle (C; n=4) receberam dieta balanceada para ganho diário de 900g; no grupo tratado com carência pronunciada de energia (CP; n=6), receberam dieta com 30 por cento a menos do nível de mantença em energia. Após 140 dias sob esses regimes de alimentação, foram coletadas amostras do fluido ruminal e urina, para realização de provas bioquímicas e funcionais, antes e às 1, 3, 6 e 9 horas após o fornecimento do alimento. A carência energética resultou em diminuição significativa na quantidade dos protozoários Entodinium, Eodinium, Isotricha, Dasytricha, Eremoplastron, Eudiplodinium, Metadinium, Charonina, Ostracodinium e Epidinium. Não houve efeito da hora de coleta sobre o total de ciliados nos grupos C e CP. Maior número de formas em divisão binária foi registrado no grupo C, na sexta e nona horas pós-alimentação (P<0,019). Observaram-se altas correlações positivas entre a contagem total de protozoários e a fermentação de glicose, amônia e o índice de excreção urinária de alantoína e negativa entre a contagem total de protozoários e a redução do azul de metileno, e correlação média entre a contagem total de protozoários e os ácidos graxos voláteis totais. A determinação da população de protozoários do rúmen é um método simples de avaliação, além de que particularmente auxilia o diagnóstico clínico da função ruminal.(AU)

Avaliação de indicadores inflamatórios no diagnóstico da mamite ovina

ABSTRACT The aim of this trial was to evaluate distinct indictors of inflammation in the diagnosis of mastitis in Santa Ines ewes. The physical examination and the dark bottom cup test were performed. Afterwards, the bacteriologic samples of 390 milk samples were analyzed. The samples were divided according to the bacteriological examination and the dark cup bottom test, resulting in 290 samples negative in both tests, 90 samples positive in the bacteriological examination and negative in the dark cup bottom test, 5 samples positive in the dark cup bottom test and negative in bacteriological culture, and 3 samples positive in both tests. The automatic somatic cell count (SSC), the California mastitis test (CMT), the pH, the chloride and lactose content, and the chloride-lactose number were established. Then, their predictive value as a indicator of inflammation was calculated assuming the bacteriologic examination as the gold standard test. The CCS, CMT and chloride content proved to be the best predictors, while the pH, the lactose content and chloride-lactose number were the least sensitive tests. The physical examination was revealed to be a nonsensitive diagnostic procedure when used as the only procedure to detect the disease, showing that other diagnostics tests are required.

RESUMO O objetivo do presente trabalho foi avaliar diferentes indicadores inflamatórios no diagnóstico da mamite em ovinos da raça Santa Inês. Após a realização do exame físico das mamas e a prova de fundo escuro, foram coletadas e analisadas 390 amostras de leite, sendo as metades mamárias divididas de acordo com o exame bacteriológico e a prova de fundo escuro, resultando em 290 amostras negativas em ambos os exames, 90 amostras negativas à prova de fundo escuro e positivas na cultura bacteriológica, 5 amostras positivas à prova de fundo escuro e negativas ao exame bacteriológico e 3 amostras positivas em ambos os testes. Nestas amostras foram realizadas a contagem automática de células somáticas (CCS), o California Mastitis Test (CMT), a determinação da concentração hidrogeniônica (pH) e dos teores de cloreto e lactose, e ainda a avaliação do índice de cloreto-lactose. Os maiores valores preditivos foram observados para a CCS, CMT e teor de cloreto; o pH, o teor de lactose e o índice cloreto-lactose apresentaram-se como marcadores inflamatórios menos sensíveis, onde se considerou o resultado da cultura bacteriológica como padrão ouro. O exame físico não se mostrou como método diagnóstico seguro quando utilizado isoladamente, exaltando a importância da associação de outros meios diagnósticos indiretos.

Ammonia poisoning causes muscular but not liver damage in cattle

Twelve steers were intraruminally administered a high dose (0.5g/kg BW) of urea to study the damage effect of ammonia poisoning on liver and/or muscles. Blood samples were collected to determine ammonia and activities of gammaGT, AST and CK. Eleven steers were successfully poisoned and treated properly, but one succumbed. Poisoned cattle showed high concentration of ammonia, and higher activities of AST and CK. The higher the ammonia, the greater were the activities of AST (r=0.59) and CK (r=0.61). The correlation between AST and CK was high and significant (r=0.80), but not between AST and gammaGT (r=0.19). The activities of AST and CK were higher after the beginning of the convulsive episodes due to ammonia poisoning. Those results showed that occurred muscle damage instead of liver damage since CK is a typical enzyme from skeletal muscle; AST is found either in skeletal muscle and hepatocytes, while gammaGT is present in hepatic cells.

Doze novilhos foram experimentalmente intoxicados por administração intrarruminal com alta dose de uréia (0,5g/kg PV) para estudar o efeito da intoxicação por uréia sobre o fígado e/ou musculatura. Foram coletadas amostras de sangue no decorrer do experimento para determinar o teor de amônia e as atividades de gama glutamiltransferase (gamaGT), aspartato aminotransferase (AST) e creatina quinase (CK). Onze novilhos apresentaram quadro clínico típico de intoxicação por amônia, porém um animal sucumbiu. Animais intoxicados apresentaram alto teor de amônia e elevada atividade de CK e AST. Quanto mais elevados foram os teores de amônia maiores foram as atividades de AST (r=0,59) e CK (0,61). A correlação entre AST e CK foi alta e significativa (r=0,80), ocorrendo o inverso com AST e gamaGT (r=0,19). Tanto a atividade de AST como de CK aumentaram significativamente a partir dos episódios convulsivos decorrentes da intoxicação por amônia. Estes resultados indicam que ocorreu uma expressiva lesão muscular, mas não hepática, visto que a CK é uma enzima típica da musculatura estriada; e o AST encontra-se tanto na musculatura estriada como nos hepatócitos, enquanto que a gamaGT está presente somente em células hepáticas.

Ammonia poisoning causes muscular but not liver damage in cattle / Intoxicação por ammonia causa lesões musculares e não hepáticas em bovinos

Twelve steers were intraruminally administered a high dose (0.5g/kg BW) of urea to study the damage effect of ammonia poisoning on liver and/or muscles. Blood samples were collected to determine ammonia and activities of gGT, AST and CK. Eleven steers were successfully poisoned and treated properly, but one succumbed. Poisoned cattle showed high concentration of ammonia, and higher activities of AST and CK. The higher the ammonia, the greater were the activities of AST (r=0.59) and CK (r=0.61). The correlation between AST and CK was high and significant (r=0.80), but not between AST and gGT (r=0.19). The activities of AST and CK were higher after the beginning of the convulsive episodes due to ammonia poisoning. Those results showed that occurred muscle damage instead of liver damage since CK is a typical enzyme from skeletal muscle; AST is found either in skeletal muscle and hepatocytes, while gGT is present in hepatic cells.(AU)

Doze novilhos foram experimentalmente intoxicados por administração intrarruminal com alta dose de uréia (0,5g/kg PV) para estudar o efeito da intoxicação por uréia sobre o fígado e/ou musculatura. Foram coletadas amostras de sangue no decorrer do experimento para determinar o teor de amônia e as atividades de gama glutamiltransferase (gGT), aspartato aminotransferase (AST) e creatina quinase (CK). Onze novilhos apresentaram quadro clínico típico de intoxicação por amônia, porém um animal sucumbiu. Animais intoxicados apresentaram alto teor de amônia e elevada atividade de CK e AST. Quanto mais elevados foram os teores de amônia maiores foram as atividades de AST (r=0,59) e CK (0,61). A correlação entre AST e CK foi alta e significativa (r=0,80), ocorrendo o inverso com AST e gGT (r=0,19). Tanto a atividade de AST como de CK aumentaram significativamente a partir dos episódios convulsivos decorrentes da intoxicação por amônia. Estes resultados indicam que ocorreu uma expressiva lesão muscular, mas não hepática, visto que a CK é uma enzima típica da musculatura estriada; e o AST encontra-se tanto na musculatura estriada como nos hepatócitos, enquanto que a gGT está presente somente em células hepáticas.(AU)

Identification of two HIV type 1 circulating recombinant forms in Brazil.

Recombination is an important way to generate genetic diversity. Accumulation of HIV-1 full-length genomes in databases demonstrated that recombination is pervasive in viral strains collected globally. Recombinant forms achieving epidemiological relevance are termed circulating recombinant forms (CRFs). CRF12_BF was up to now the only CRF described in South America. The objective was to identify the first CRF in Brazil conducting full genome analysis of samples sharing the same partial genome recombinant structure. Ten samples obtained from individuals residing in Santos, Brazil, sharing the same recombination pattern based on partial genome sequence data, were selected from a larger group to undergo full length genome analysis. Near full length genomes were assembled from overlapping fragments. Mosaic genomes were evaluated by Bootscan, alignment inspection, and phylogenetic analysis using neighbor joining and maximum likelihood. Full genomes were also analyzed by split decomposition. We were able to identify five mosaic genomes. Two of these structures were represented by at least three samples derived from epidemiologically unlinked individuals. These structures were named CRF28_BF and CRF29_BF and are the second and third CRFs composed exclusively by subtypes B and F as well as the second and third CRFs encountered in South America. Other recombinant forms studied here resembled CRF28_BF and CRF29_BF. Our results suggest that a diverse population of related recombinants, including CRFs may play an important part in the Brazilian and South American epidemic.

Cyclosporine bioequivalence study: quantification using fluorescence polarization immunoassay (FPIA) and radioimmunoassay (RIA).

OBJECTIVE: The aim of study was to compare the bioavailability of 2 cyclosporine capsule formulations (100 mg; Sigmasporin Microoral from Novaquímica Divisão Nature's Plus Farmacêutica Ltd., Brazil, as test formulation and Sandimmune Neoral from Novartis Biociências S.A., Brazil, as reference formulation) in 24 healthy male volunteers. METHODS: The study was open, randomized, with a 2-period crossover, a 1-week washout interval between doses. Blood samples were obtained over a 12-hour interval after each oral administration of cyclosporine (2 capsules of 100 mg of each formulation). Cyclosporine blood concentrations were quantified using a fluorescence polarization immunoassay (FPIA) method provided by Abbott Axsym System and Cyclo-Trac SP. Whole-blood radioimmuoassay (RIA) kit was provided by DiaSorin. These assays provided concentration-time curves for cyclosporine in blood concentration from which the following pharmacokinetic parameters were obtained: AUC(last), AUC(inf), Cmax. RESULTS: Geometric mean and 90% confidence intervals (CI) of Microoral/Neoral as percent ratios were 94.5% (90.8-98.4%) for AUC(last), 93.8% (89.7-98.1%) for AUC(inf), and 98.1% (94.5-101.8%) for Cmax when cyclosporine was determined using FPIA and 96.1% (91.9 to 100.6%) for AUC(last), 95.2% (90.2-100.5%) for AUC(inf), and 99.4% (96.4-102.4%) for Cmax using RIA. CONCLUSION: Since the 90% CI for Cmax, AUC(last) and AUC(inf) ratio were within the 80-125% interval proposed by US-FDA, it is concluded that Sigmasporin Microoral 100 mg capsule formulation is bioequivalent to Sandimmune Neoral 100 mg capsule formulation with regard to both rate and the extent of absorption.

Quantification of nimesulide in human plasma by high-performance liquid chromatography/tandem mass spectrometry. Application to bioequivalence studies.

A method based on liquid chromatography with negative ion electrospray ionization and tandem mass spectrometry is described for the determination of nimesulide in human plasma. Liquid-liquid extraction using a mixture of diethyl ether and dichloromethane was employed and celecoxib was used as an internal standard. The chromatographic run time was 4.5 min and the weighted (1/x) calibration curve was linear in the range 10.0-2000 ng x ml(-1). The limit of quantification was 10 ng x ml(-1), the intra-batch precision was 6.3, 2.1 and 2.1% and the intra-batch accuracy was 3.2, 0.3 and 0.1% for 30, 300 and 1200 ng x ml(-1) respectively. The inter-batch precision was 2.3, 2.8 and 2.7% and the accuracy was 3.3, 0.3 and 0.1% for 30, 300 and 1200 ng x ml(-1) respectively. This method was employed in a bioequivalence study of one nimesulide drop formulation (nimesulide 50 mg x ml(-1) drop, Medley S/A Indústria Farmacêutica, Brazil) against one standard nimesulide drop formulation (Nisulid, 50 mg x ml(-1) drop, Astra Médica, Brazil). Twenty-four healthy volunteers (both sexes) took part in the study and received a single oral dose of nimesulide (100 mg, equivalent to 2 ml of either formulation) in an open, randomized, two-period crossover way, with a 2-week washout interval between periods. The 90% confidence interval (CI) for geometric mean ratios between nimesulide and Nisulid were 93.1-109.6% for C(max), 87.7-99.8% for AUC(last) and 88.1-99.7% for AUC(0-infinity). Since the 90% CI for the above-mentioned parameters were included in the 80-125% interval proposed by the US Food and Drug Administration, the two formulations were considered bioequivalent in terms of both rate and extent of absorption.

Significance of isolated hepatitis B core antibody in blood donors from São Paulo.

The clinical significance of isolated anti-HBc is still a challenge. To elucidate the real importance of this finding in our blood donors, an investigation algorithm was tested. One hundred and twelve isolated anti-HBc seropositive blood donors underwent clinical evaluation and retesting of HBV markers. Those who presented repeatedly reactive isolated anti-HBc, received a single dose of hepatitis B recombinant vaccine to verify anti-HBs early response. A HBV-DNA determination by PCR was done for those who did not test positive to anti-HBs after vaccine. The level of anti-HBc was recorded as a ratio of the sample-to-cut-off values (S:C ratio) in 57 candidates at donation. Comparing true and false-positive anti-HBc results, the different S:C ratios of them were statistically significant and when less than 2, implying in a false-positive result probability over 80%. A high percent of false-positive results (16.07%) was verified after anti-HBc retesting. HBV immunity was characterized in 49.11%, either by anti-HBs detection in retesting (15.18%), or after a single dose HBV vaccination (33.93%). HBV-DNA was negative in all tested donors. In conclusion, this algorithm was useful to clarify the meaning of isolated anti-HBc in most of our blood donors.

Comparative bioavailability of 4 amoxicillin formulations in healthy human volunteers after a single dose administration.

OBJECTIVE: To compare the bioavailability of two amoxicillin oral suspension (250 mg/5 ml) formulations and two amoxicillin capsule (500 mg) formulations (Amoxicilina from Medley S/A Indústria Farmaceûtica, Brazil, as test formulations and Amoxil from SmithKline Beecham Laboratórios Ltda., Brazil, as reference formulations) in 48 volunteers of both sexes. MATERIAL AND METHODS: The study was conducted open with a randomized two-period crossover design and a one-week washout period. Plasma samples were obtained over a 12-hour interval. Amoxicillin concentrations were analyzed by combined reversed phase liquid chromatography and tandem mass spectrometry (LC-MS-MS) with positive ion electrospray ionization using the selected ion monitoring method. From the amoxicillin plasma concentration vs. time curves the following pharmacokinetic parameters were obtained: AUC(last), AUC(0-infinity) and Cmax. RESULTS: Geometric mean of Amoxicilina/Amoxil 250 mg/5 ml individual percent ratio was 103.70% for AUC(last), 103.15% for AUC(0-infinity) and 106.79% for Cmax. The 90% confidence intervals were 97.82-109.94%, 97.40 to 109.24%, and 96.38-118.33%, respectively. Geometric mean of Amoxicilina/Amoxil 500 mg capsule individual percent ratio was 93.26% for AUC(last), 93.27% for AUC(0-infinity) and 90.74% for Cmax. The 90% confidence intervals were 85.0-102.33%, 85.12-102.31%, and 80.14-102.73%, respectively. CONCLUSION: Since the 90% CI for both Cmax, AUC(last) and AUC(0-inifnity) were within the 80-125% interval proposed by the Food and Drug Administration, it was concluded that Amoxicilina 250 mg/5 ml oral suspension and Amoxicilina 500 mg capsule were bioequivalent to Amoxil 250 mg/5 ml oral suspension and to Amoxil capsule 500 mg, respectively, with regard to both the rate and extent of absorption.

Accumulative sodium poisoning in Brazilian swine fed whey.

Fifty 3-4-mo-old piglets died of accumulative sodium poisoning, but none of the 60 adult pigs with the same feeding and management did. The average ambient temperature throughout the period was 32 C. The herd had been regulary fed whey, ground corn and vegetables, but for at least 2 d the pigs were deprived of water and then water was offered ad libitum. Twenty hours later 20 piglets had died and the remaining exhibited classical nervous signs and died within 14 h. Only the piglets had a high degree of dehydration caused by water deprivation and exacerbated by the high average ambient temperature. Pulmonary edema was evident in most piglets. Acute cerebral edema and meningoencephalitis were present in all animals, but there was no polioencephalomalacia. The sodium accumulation was caused by the continuing intake of whey which contained 10,810 mg sodium/L. To produce the whey, 2 kg of sodium chloride had been added to every 100 kg of milk.