Tillering dynamics of Alexandergrass pasture under nitrogen fertilization / Dinâmica do perfilhamento da pastagem de Papuã sob fertilização nitrogenada

This study was carried out to evaluate the effect of nitrogen fertilization on tillering dynamics of Alexandergrass (Urochloa plantaginea (Link) Hitch) grazed by beef heifers in a rotational stocking grazing method. The experiment was conducted in a completely randomized design following a repeated measure arrangement, three levels of nitrogen (Zero, 150 and 300kg ha-1) and two area repetition. The classification of tillers into categories (basal and axillary) showed a greater number of basal tillers, due to their higher appearance rate, in the absence of nitrogen fertilization. Nitrogen fertilization promoted higher axillary tiller density. The use of nitrogen promotes the renewal of axillary tillers of Alexandergrass, without compromising the stability of the tiller population.(AU)

Esse estudo foi conduzido para avaliar o efeito da fertilização nitrogenada no perfilhamento do Papuã (Urochloa plantaginea (Link) Hitch) pastejado por novilhas de corte em um método de lotação rotacionada. O delineamento foi inteiramente casualizado com arranjo de medidas repetidas, três níveis de nitrogênio (Zero, 150 e 300kg ha -1 ) e duas repetições de área. A classificação dos perfilhos em categorias (basais e axilares) mostrou um maior número de perfilhos basais, devido à sua maior taxa de aparecimento, na ausência de adubação nitrogenada. A fertilização nitrogenada promoveu maior densidade de perfilhos axilares. O uso de nitrogênio promove renovação dos perfilhos axilares no Papuã, sem comprometer a estabilidade da população de plantas.(AU)

Análise bioeconômica do farelo de arroz na recria de bezerras de corte em azevém / Bioeconomics analyses of rice brain use for reaning of beef heifers on Italian ryegrass pasture

Objetivou-se avaliar a economicidade, o desempenho produtivo e o desenvolvimento das estruturas corporais relacionadas ao trato reprodutivo de bezerras de corte mantidas exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou em pastagem de azevém recebendo 0,5 e 1,0% do peso corporal (PC) de farelo de arroz integral (FAI) como suplemento. O método de pastejo foi o rotativo, e o intervalo entre pastejos foi estabelecido considerando-se a soma térmica necessária para a emissão de 1,5 folha de azevém (187,5ºC). O delineamento experimental foi o inteiramente ao acaso, com medidas repetidas no tempo. O ganho médio diário das bezerras não diferiu entre os sistemas alimentares. A taxa de lotação e o ganho de peso por área foram maiores quando as bezerras receberam 1,0% do PC de farelo de arroz integral. A maior margem bruta foi observada quando as bezerras receberam 0,5% de FAI. O custo variável obtido para os diferentes sistemas alimentares apresentou comportamento crescente em função dos níveis de FAI, superando o uso exclusivo do azevém em 49,4% e 81,8%. O peso corporal final, o escore de condição corporal e o escore do trato reprodutivo não diferiram entre os sistemas alimentares. Considerando-se um animal adulto com peso corporal médio de 450kg, as bezerras ao início do experimento apresentaram 34% do peso corporal adulto e peso corporal final médio de 56% do peso adulto. A utilização de níveis de farelo de arroz integral (FAI) na recria de bezerras de corte sob pastejo rotativo em azevém não modifica o ganho médio diário e o escore do trato reprodutivo. O fornecimento de 1% de FAI mostrou efeito substitutivo, proporcionando um incremento de 31,2% na taxa de lotação e de 38,3% no ganho por área. O sistema alimentar AZ0,5 permitiu uma maior margem bruta; já o retorno financeiro direto foi positivo em todos os sistemas alimentares, com melhor retorno calculado para o uso exclusivo do azevém.(AU)

The experiment was carried out with the objective of evaluating the gross margin, the productive and reproductive performance of beef heifers kept exclusively on ryegrass pasture (Lolium multiflorum Lam.) or ryegrass receiving 0.5 and 1.0% of body weight (BW) of whole rice bran as supplement. The grazing method was the rotational and the interval between grazings was set by the thermal sum required for the appearing of 1.5 ryegrass leaf (187.5ºC). The experimental design was completely randomized with repeated measures. Heifers average daily gain did not differ between the feeding systems. The stocking rate and weight gain per area were higher when heifers received 1.0% BW of whole rice bran. The higher gross margin was observed when heifers received 0.5% BW. The variable cost obtained for the different feeding systems showed increasing behavior depending on the WRB levels, surpassing the exclusive use of ryegrass in 49.4% and 81.8%. The final body weight, body condition score and reproductive tract score did not differ between the feeding systems. Considering an adult animal with an average body weight of 450kg, the heifers at the beginning of the experiment showed 34% of mature body weight and final average body weight of 56% of adult weight. The use of rice bran levels (FAZ) in the rearing of beef heifers under rotational grazing on ryegrass does not change the average daily gain and the reproductive tract score. The supply of 1% FAI showed substitutive effect, providing an increase in capacity of 31,2% and 38.3% rate of the gain per area. The food system AZ0,5 allowed a higher gross margin, as the economic return was positive in all food systems, with better return calculated for the exclusive use of ryegrass.(AU)

Coinfecções por Leishmania infantum, Neospora caninum e Toxoplasma gondii em cães necropsiados da região central do Rio Grande do Sul, Brasil / Coinfections by Leishmania infantum, Neospora caninum and Toxoplasma gondii in necropsied dogs from the central region of Rio Grande do Sul, Brazil

O objetivo do presente trabalho foi determinar a presença de anticorpos para Leishmania infantum, Neospora caninum e Toxoplasma gondii, por meio da reação de imunofluorescência indireta (RIFI), em cães (n=78) provenientes da região central do Rio Grande do Sul, necropsiados no Hospital Veterinário da Universidade Federal de Santa Maria (UFSM), bem como avaliar os dados epidemiológicos, sazonais e anátomo-histopatológicos. Do total de animais avaliados, 67,9% (53/78) apresentaram soropositividade para ao menos um agente. A ocorrência de anticorpos para L. infantum, N. caninum e T. gondii foi de 33,3 (26/78), 37,1 (29/78) e 43,5% (34/78), respectivamente. Detectaram-se monoinfecções em 9,4% (5/53) para L. infantum, 18,8% (10/53) para N. caninum e 20,7% (11/53) para T. gondii. As coinfecções foram observadas em 27/53 (50,9%) dos animais. As infecções ocorreram independentemente de idade, sexo, procedência ou raça (P>0,05). Não se verificaram lesões anatomo-histopatológicas relacionadas aos agentes pesquisados, caracterizando-os como animais assintomáticos. Os resultados confirmaram a exposição de cães a esses protozoários na região central do RS e, em especial, demonstraram a circulação do agente causador da leishmaniose em uma área considerada indene para a enfermidade.(AU)

The present paper is aimed to determine the presence of antibodies for Leishmania infantum, Neospora caninum and Toxoplasma gondii by indirect immunofluorescence (IIF) in dogs (n=78) from the central region in the state of Rio Grande do Sul necropsied in the Veterinary Hospital from Universidade Federal de Santa Maria (UFSM). The data was evaluated regarding epidemiological, anatomic, and histopathologic findings. Of the total animals evaluated, 67.9% (53/78) showed seropositivity for at least one agent. The occurrence of antibodies to L. infantum, N. caninum and T. gondii was 33.3% (26/78) 37.1% (29/78) and 43.5% (34/78), respectively. The mono infections were detected in 9.4% (5/53) of L. infantum, 18.8% (10/53) for N. caninum and 20.7% (11/53) T. gondii. The coinfections occurred in 50.9% (27/53) of animals. There were not anatomical and histopathological lesions regarding these surveyed agents, characterizing them as subclinical animals. The results confirmed the exposition of dogs to these protozoa in the central region of the RS, highlighting the circulation of the causer agent of leishmaniasis in an area considered harmless for the disease.(AU)

Desempenho de bezerras de corte em pastagem de azevém recebendo farelo de arroz com ou sem monensina / Beef heifers performance in ryegrass pasture supplemented with rice bran with or without monensin

Foi estudado o desempenho de bezerras de corte em pastagem de azevém (Lolium multiflorum Lam.), com animais exclusivamente em pastejo ou recebendo farelo de arroz integral (FAI) em nível de 0,8% do peso corporal, associado ou não à monensina. O método de pastejo foi contínuo com número variável de animais. O delineamento experimental foi o inteiramente ao acaso, com medidas repetidas no tempo, com três tratamentos e três repetições de área. O uso de FAI associado ou não à monensina não expressou alteração na área pélvica das bezerras, na taxa de lotação e no ganho de peso por área. As bezerras que receberam FAI, com ou sem monensina, apresentaram maior ganho diário de peso corporal, peso corporal, escore de condição corporal e relação peso corporal:altura. O fornecimento de suplementação energética para bezerras de corte é uma alternativa viável em sistemas de produção que visam à redução da idade ao primeiro acasalamento.(AU)

The experiment was carried out with the objective of evaluating beef cattle heifer performance when exclusively grazing Italian ryegrass (Lolium multiflorum Lam.) pasture or supplemented with whole rice bran (WRB), equivalent to 0.8% of body weight, associated or not with monensin. The grazing method was "put-and-take" stocking, in a completely randomized experimental design, with replicated measures over time, with three treatments and three replications of area. The use of WRB, associated or not with monensin, did not affect heifers' pelvic area, stocking rate and area weight gain. Animals fed WRB, with or without monensin, showed greater average daily gain, body weight, body condition score, and body weight:height ratio. Energetic supplementation for beef heifers is one viable option in production systems seeking age reduction at the first mating.(AU)

Forage intake and performance of heifers fed rice bran on Alexandergrass pasture / Ingestão de forragem e performance de bezerros alimentados com farelo de arroz integral em pasto de papuã

he study was carried out to evaluate the forage intake and performance of beef heifers from 15 to 18 months of age, grazing on Alexandergrass (Urochloaplantaginea (Link) Hitch) fed rice bran (RB; 0, 0.5 and 1.0% body weight (BW)). The experiment consisted of a completely randomized design with repeated measurement arrangement. Rice bran increased the total dry matter (DM) intake, total crude protein intake and total digestible nutrients intake. Heifers that received 1.0% BW of RB reduced forage DM intake, increasing by 18% stocking rate (SR) and by 27% BW gain per area in comparison with the system exclusively on pasture. In the system in which the heifers were fed 0.5% BW of RB, SR and gain per area were similar to other systems. In this feeding system, we observed average daily gain 18% higher than when heifers were exclusively on pasture and gain similar to that observed when using 1.0% BW of RB. In Alexandergrass pasture it is recommended to supply 0.5% BW of RB for beef heifers from 15 to 18 months of age, to promote better productive responses compared to heifers exclusively on pasture and similar to those observed when supplying 1.0% BW of RB.(AU)

O presente trabalho foi desenvolvido com o objetivo de avaliar o consumo de forragem e o desempenho de novilhas de corte dos 15 aos 18 meses de idade, em pastejo em papuã (Urochloaplantaginea(Link) Hitch), recebendo farelo de arroz integral (FAI) como suplemento (0, 0,5 e 1,0% do peso corporal (PC)). O delineamento experimental foi o inteiramente ao acaso, com medidas repetidas no tempo. O uso de FAI aumentou o consumo total de matéria seca (MS), o consumo total de proteína bruta e de nutrientes digestíveis totais em relação às novilhas exclusivamente em pastejo. As novilhas que receberam 1,0% do PC de FAI reduziram o consumo de MS da forragem, aumentando em 18% a taxa de lotação (TxLot) e em 27% o ganho de PC por hectare (GPA) em comparação com o sistema exclusivamente a pasto. No sistema em que as novilhas receberam 0,5% do PC de FAI, a TxLot e o GPA foram similares aos demais sistemas. Nesse sistema alimentar, foi observado GDM 18% superior em relação às novilhas exclusivamente em pastejo e ganho similar ao uso de 1,0% do PC de FAI. Em pastagem de papuã, é recomendado o uso de 0,5% do PC de FAI para novilhas de corte dos 15 aos 18 meses de idade, por promover respostas produtivas superiores em relação às novilhas exclusivamente em pastejo e semelhantes às observadas quando fornecido 1,0% do PC de FAI.(AU)

Padrões de deslocamento de bezerras de corte recebendo suplemento em pastagem de azevém / Displacement patterns of beef heifers receiving supplement in Italian ryegrass pasture

O objetivo deste trabalho foi avaliar o tempo de permanência, o número de bocados nas estações e o deslocamento entre estações alimentares por bezerras de corte em pastagem de azevém (Lolium multiflorum Lam.) e recebendo farelo de arroz integral como suplemento em três níveis (0,0; 0,5 e 1,0% PC). O método de pastejo foi rotacionado, com número variável de animais. O período de descanso foi estabelecido pela soma térmica (ST) de 187,5ºC, equivalente à ST necessária para a emissão de 1,5 folhas de azevém. O delineamento experimental foi inteiramente ao acaso, com medidas repetidas no tempo, três níveis de suplemento e seis ciclos de pastejo. O tempo de permanência em cada estação alimentar (9,4s) foi semelhante para bezerras exclusivamente a pasto e as que receberam 1,0% PC de suplemento. As bezerras que receberam 0,5% PC de suplemento permanecem 1,1s a mais na estação alimentar quando comparadas com as bezerras nos demais níveis. O deslocamento entre estações alimentares (1,7 passos) e o número de bocados em cada estação alimentar (sete bocados) foram semelhantes independentemente do nível de farelo de arroz fornecido. As bezerras, ao receberem 0,5% PC de farelo de arroz, são mais dependentes da presença de lâminas foliares no estrato pastejável do dossel.(AU)

The aim of this study was to evaluate the length of stay and number of bites in the feeding stations and displacement between feeding stations for beef heifers on ryegrass pasture (Lolium multiflorum Lam.) and receiving rice bran as a supplement in three levels (0.0; 0.5 and 1.0% BW). The grazing method was rotational with variable number of animals. The interval between grazing was set by the thermal sum (TS) of 187.5ºC equivalent to TS necessary for externalization of 1.5 leaf lamina of ryegrass. The experimental design was completely randomized with repeated measures on time, three levels of supplement and six grazing periods. The stay time at each feeding station (9.4 sec.) was similar for heifers exclusively on pasture and receiving 1.0% supplement BW. Heifers that received 0.5% supplement BW remained 1.1 sec. longer at the feeding station than heifers on other supplement levels. Displacement between feeding stations (1.7 steps) and the number of bites in each feeding station (7 bites) were similar regardless of the supplied rice bran level. Heifers receiving 0.5% rice bran BW are more dependent on the presence of leaf blades in grazeable stratum of canopy.(AU)

Uso de suplementos para acasalamento de bezerras Angus aos 14 meses de idade / Use of supplements for mating Angus heifers at 14 months

O experimento foi conduzido com o objetivo de avaliar o desenvolvimento de novilhas de corte em três sistemas alimentares: exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou em pastagem de azevém e recebendo 0,93% do peso corporal de grão de milho inteiro ou grão de aveia branca como suplemento. O delineamento experimental foi o inteiramente ao acaso, com medidas repetidas no tempo. A utilização da pastagem de azevém, com ou sem uso de grão de milho ou aveia como suplemento, possibilita desenvolvimento reprodutivo para que as bezerras sejam acasaladas aos 14 meses de idade. O peso corporal ao início da utilização da pastagem é determinante para que as bezerras atinjam adequado desenvolvimento reprodutivo no início da estação de acasalamento.(AU)

The experiment was carried out to evaluate the development of beef heifers in three feeding systems: exclusively on ryegrass pasture (Lolium multiflorum Lam.) or in ryegrass pasture and receiving 0.93% of body weight of corn whole grain or oats grain as a supplement. The experimental design was completely randomized with repeated measures in time. The utilization of ryegrass pasture, with or without the use of corn or oats grain as a supplement, provides reproductive development of heifers in order for them to be mated at 14 months of age. The body weight at the beginning of pasture utilization is crucial for heifers to reach adequate reproductive development at the beginning of the mating season.(AU)

Canopy structure, ingestive behavior and displacement patterns of beef heifers grazing warm-season pastures / Estrutura do dossel, comportamento ingestivo e padrões de deslocamento de novilhas de corte em pastagens de estação quente

Analyses in a data set of six experiments (n=436) was performed with the aim of characterizing canopy structure and forage intake patterns of beef heifers grazing on Pearl millet (Pennisetum americanum), Alexandergrass (Urochloa plantaginea) and Coastcross (Cynodon dactylon). Forage and leaf lamina mass were similar among species, 3001.4 and 668.1kg of DM/ha, respectively, while leaf:stem ratio of canopy vertical strata was different. Intake rate (12g DM/minute), bite mass (0.343g DM/bite) and bite rate (36.6bites per minute) were similar in Pearl millet and Alexandergrass. In Pearl millet, neutral detergent fiber content (56.1%) in forage as grazed, grazing time (518.9 minutes/day) and displacement rate (8.8 steps/minute) were lower. Increased grazing time (639 minutes/day) represented the main compensatory mechanism for smaller bite masses (0.234g DM/bite) and reduced intake rates (8.8 grams of DM/minutes) in Coastcross. Bite rate variations happened as a response to constraints imposed by canopy structure. Surface utilization on Alexandergrass and Coastcross was increased by greater number of feeding stations visited and displacement rate. Ingestive behavior components of beef heifers are affected by differences in the canopy structure of Pearl millet, Alexandergrass and Coastcross.

Foi realizada a análise conjunta dos resultados de seis experimentos (n=436) com o objetivo de caracterizar a estrutura do dossel e os padrões de ingestão de forragem por novilhas de corte em pastagem de milheto (Pennisetum americanum), papuã (Urochloa plantaginea) e coastcross (Cynodon dactylon). As massas de forragem e de lâminas foliares foram similares entre as três espécies forrageiras, 3001,4 e 668,1kg/ha de MS, respectivamente, enquanto a relação lâmina:colmo do perfil vertical do dossel diferiu. A taxa de ingestão (12g MS/minuto), a massa de bocados (0,343g MS/bocado) e a taxa de bocados (36,6 bocados/minuto) foram similares no milheto e no papuã. O conteúdo de fibra em detergente neutro (56,1%) na forragem da simulação do pastejo, o tempo de pastejo (518,9 minutos/dia) e a taxa de deslocamento (8,8 passos/minuto) foram menores no milheto. O incremento no tempo de pastejo (639 minutos/dia) representou o mecanismo compensatório para a menor massa de bocados (0,234g MS/bocado) e para a reduzida taxa de ingestão (8,8 gramas MS/minuto) no coastcross. Variações na taxa de bocado ocorrem como resposta às limitações impostas pela estrutura do dossel. A exploração da área da pastagem de papuã e de coastcross foi aumentada pelo maior número de estações alimentares visitadas e pela maior taxa de deslocamento. Os componentes do comportamento ingestivo de novilhas de corte são afetados por diferentes estruturas no dossel de milheto, papuã e coastcross.

Cardiac toxoplasmosis after heart transplantation diagnosed by endomyocardial biopsy.

We describe a case of cardiac toxoplasmosis diagnosed by routine endomyocardial biopsy in a patient with trimethoprim-sulfamethoxazole (TMP-SMX) intolerance on atovaquone prophylaxis. Data are not available on the efficacy of atovaquone as Toxoplasma gondii prophylaxis after heart transplantation. In heart transplant patients in whom TMP-SMX is not an option, other strategies may be considered, including the addition of pyrimethamine to atovaquone.

Atividade de nanoformulações de Melaleuca alternifolia e terpinen-4-ol em isolados de Rhodococcus equi / Melaleuca alternifolia activity in nanoformulations and terpinen-4-ol against Rhodococcus equi isolates

Rhodococcus equi é o agente etiológico da rodococose equina, importante doença respiratória de potros. Especialmente na última década, a emergência de cepas resistentes aos antimicrobianos empregados no tratamento da rodococose tem sido relatada. Nesse sentido, há a necessidade de estudos envolvendo terapias alternativas e novas tecnologias, incluindo o uso de plantas medicinais e nanotecnologia. Neste trabalho utilizou-se Melaleuca alternifolia nas seguintes formulações: óleo livre, nanocápsula, nanoemulsão e a combinação de óleo livre com nanocápsula e com nanoemulsão, além do seu composto majoritário, terpinen-4-ol, a fim de verificar a atividade antimicrobiana frente a isolados de R. equi de diferentes origens. Utilizou-se o método de microdiluição em caldo na determinação das concentrações inibitória mínima (CIM) e bactericida mínima (CBM) das diferentes formulações frente aos isolados (n=24). Verificou-se baixo potencial para atividade antibacteriana de M. alternifolia na formulação de óleo livre. Todavia, essa atividade foi potencializada quando se incorporou o óleo essencial às nanoformulações. O composto terpinen-4-ol demonstrou potencial atividade antibacteriana quando incorporado ao óleo essencial e quando utilizado isoladamente. Verificou-se que tanto M. alternifolia quanto terpinen-4-ol testados possuem atividade antimicrobiana contra isolados de R. equi, sugerindo seu emprego em estudos avaliando seu potencial para o tratamento da rodococose.

Rhodococcus equi causes rodococose in horses, characterized by bronchopneumonia in foals. Due to reports of antimicrobial resistance, it is important to develop studies involving alternative therapies and new technologies, including the use of medicinal plants and nanotechnology. In this work, the plant Melaleuca alternifolia in oil free formulations, nanocapsule, nanoemulsion and the combination of free and nanocapsule oil nanoemulsion, besides its major compound, terpinen-4-ol, were used in order to verify antimicrobial activity against isolates of R. equi. The broth microdilution method was employed to determine the minimum inhibitory (MIC) and minimum bactericidal (MBC) concentrations of different formulations against 24 isolates. There was low antibacterial activity of M. alternifolia in oil free formulation; however, it was observed that the activity was enhanced when incorporated as essential oil the nanoformulations. The major compound, terpinen-4-ol, showed bactericidal and bacteriostatic activity when used alone. It is suggested that M. alternifolia, in association with nanocarriers systems, as well as terpinen -4-ol, presents potential for future studies concerning the equine rodococosis therapy.

Padrões de deslocamento de bezerras de corte que receberam suplementos isolipídicos em pastagem de azevém / Displacement patterns of beef heifers receiving iso lipidic supplements on ryegrass pasture

Foram avaliados o uso de estações alimentares, o deslocamento e a taxa de bocado de bezerras de corte que foram mantidas exclusivamente em pastagem de azevém (Lolium multiflorum Lam.) ou receberam suplementos (grão de milho ou gordura). O método de pastejo foi contínuo, com número variável de animais. O delineamento experimental foi inteiramente ao acaso, com medidas repetidas no tempo. O número de bocados realizados por estação alimentar é similar quando as bezerras recebem suplemento ou não. As equações de predição mostram que a massa de lâminas foliares exerce maior influência no tempo por estação alimentar quando as bezerras permanecem exclusivamente em pastejo de azevém ou recebem grão de milho como suplemento, enquanto para bezerras que recebem gordura, essa variável é influenciada pela oferta de forragem. O deslocamento (passos entre estações e passos por minuto) de bezerras exclusivamente em pastejo e que recebem grão de milho é influenciado pela estrutura e qualidade do pasto. A taxa de bocado dos animais suplementados com grão de milho e gordura é influenciada pela proporção de lâminas foliares no dossel. Equações de regressão múltipla, considerando-se os atributos do pasto e da pastagem, podem ser utilizadas como modelos preditores do uso de estações alimentares, deslocamento e taxa de bocado de bezerras de corte...

The use of feeding stations, displacement patterns and bite rate of beef heifers kept exclusively on Italian ryegrass (Lolium multiflorum Lam) or receiving supplements (corn grain or fat) were evaluated. The grazing method was continuous with a variable number of animals. The experimental design was a completely randomized design with repeated time measures. The number of bites per feeding station is similar when heifers receive supplement or not. The prediction equations show that the mass of leaf lamina has greater influence on the time per feeding station when heifers remain exclusively grazing ryegrass or receive corn grain as a supplement, while for calves that receive fat this variable is influenced by forage allowance. The displacement (steps between stations and steps per minute) of heifers exclusively on ryegrass or receiving corn grain is influenced by the structure and quality of grass. The bite rate of heifers supplemented with corn grain and fat is influenced by the proportion of leaf lamina in the sward. Multiple regression equations considering the attributes of grass and pasture can be used as models to predict the use of feeding stations, displacement and bite rate of beef heifers...

BRAF, NRAS and HRAS mutations in spitzoid tumours and their possible pathogenetic significance.

BACKGROUND: The relationships between so-called spitzoid tumours have proven difficult to understand. OBJECTIVES: To address three questions: does spitzoid tumour morphological similarity reflect molecular similarity? Does Spitz naevus progress into spitzoid melanoma? Are ambiguous spitzoid tumours genuine entities? METHODS: BRAF, NRAS and HRAS mutations were analysed using single-strand conformational polymorphism analysis and sequencing. RESULTS: Both Spitz naevi and spitzoid melanoma had a lower combined BRAF and NRAS mutation frequency compared with common acquired naevi (P = 0.0001) and common forms of melanoma (P = 0.0072), respectively. To look for evidence of progression from Spitz naevi to spitzoid melanoma, HRAS was analysed in 21 spitzoid melanomas, with no mutations identified. The binomial probability of this was 0.03 based on an assumption of a 15% mutation frequency in Spitz naevi with unbiased progression. Under these assumptions, HRAS mutations must be rare/absent in spitzoid melanoma. Thus, Spitz naevi seem unlikely to progress into spitzoid melanoma, implying that ambiguous spitzoid tumours cannot be intermediate degrees of progression. In addition, the data suggest that HRAS mutation is a potential marker of benign behaviour, in support of which none of three HRAS mutant spitzoid cases metastasized. CONCLUSIONS: First, the morphological similarity of spitzoid tumours reflects an underlying molecular similarity, namely a relative lack of dependence on BRAF/NRAS mutations. Second, Spitz naevi do not appear to progress into spitzoid melanoma, and consequently ambiguous spitzoid tumours are likely to be unclassifiable Spitz naevi or spitzoid melanoma rather than genuine entities. Third, HRAS mutation may be a marker of Spitz naevus, raising the possibility that other molecular markers for discriminating Spitz naevi from spitzoid melanoma can be discovered.

A novel polymorphism in the 1A promoter region of the vitamin D receptor is associated with altered susceptibilty and prognosis in malignant melanoma.

The association of Taq 1 and Fok 1 restriction fragment length polymorphisms of the vitamin D receptor with occurrence and outcome of malignant melanoma (MM), as predicted by tumour (Breslow) thickness, has been reported previously. We now report a novel adenine-guanine substitution -1012 bp relative to the exon 1a transcription start site (A-1012G), found following screening by single-stranded conformational polymorphism of this promoter region. There was a total of 191 MM cases, which were stratified according to conventional Breslow thickness groups, cases being randomly selected from each group to form a distribution corresponding to the known distribution of Breslow thickness in our area, and this population (n=176) was compared to 80 controls. The A allele was over-represented in MM patients and, with GG as reference, odds ratio (OR) for AG was 2.5, 95% confidence interval (CI) 1.1-5.7, (P=0.03) and AA 3.3, CI 1.4-8.1, (P=0.007). The outcome was known in 171 of 191 patients and the A allele was related to the development of metastasis, the Kaplan-Meier estimates of the probability of metastasis at 5 years being: GG 0%; AG 9%, CI 4-16%; AA 21%, CI 12-36%; (P=0.008), and to thicker Breslow thickness groups (P=0.04). The effect on metastasis was independent of tumour thickness and A-1012G may have predictive potential, additional to Breslow thickness. Neither the Fok 1 nor Taq 1 variants (f and t) were significantly related to the development of metastasis, although there was a strong relationship of fftt with the thickest Breslow thickness group (P=0.005). There was an interaction between the A-1012G and Fok 1 polymorphisms (P=0.025) and the Fok 1 variant enhanced the effect of the A allele of the A-1012G polymorphism on metastasis, the probability of metastasis for AAff at 5 years follow-up being 57%, CI 24-92%.

Nuclear beta-catenin in basal cell carcinoma correlates with increased proliferation.

BACKGROUND: Virtually all BCCs have deregulation of the Hedgehog (Hh) signalling pathway and a proportion show nuclear beta-catenin accumulation. The latter is thought to be due to Hh pathway-directed Wnt expression but this has not been tested. An alternative cause of nuclear beta-catenin accumulation is gene mutation, which stabilizes the protein. Theoretically, reduced E-cadherin expression could also be important because it can sequester beta-catenin at the cell membrane. In turn, nuclear beta-catenin can increase expression of MYC and cyclin D1, thus potentially altering proliferation. OBJECTIVES: To assess whether nuclear beta-catenin occurs in BCC, and to look at potential causes and consequences. METHODS: Nuclear beta-catenin was assessed by immunohistochemistry, and its causes by analysis of E-cadherin expression, beta-catenin exon 3 mutation and WNT5A expression. Its consequences were assessed by analysing proliferation. RESULTS: We found nuclear beta-catenin in 20 of 86 paraffin-embedded sections of BCCs using immunohistochemistry. BCCs showed increased WNT5A relative to the surrounding skin. No mutations in exon 3 of the beta-catenin gene were found in 10 cases. There was no association between beta-catenin localization and E-cadherin expression. Tumours with nuclear beta-catenin had significantly higher proliferation (P < 0.01). CONCLUSIONS: The absence of beta-catenin gene mutations indicate that the Hh pathway-directed Wnt signalling remains the most likely cause of nuclear beta-catenin accumulation in BCC. Additionally, the correlation with increased proliferation is the first evidence that nuclear beta-catenin may have a biological effect. However, a causal link between Hh pathway deregulation, Wnt ligand overexpression, nuclear beta-catenin accumulation and increased proliferation remains to be confirmed.

Flap monitoring after head and neck reconstruction: evaluating an observation protocol.

This study evaluated the postoperative free-flap monitoring frequency protocol used in a maxillofacial unit for patients receiving free-tissue transfer for reconstruction following orofacial cancer. All free-tissue transfers undertaken in the unit between January 1992 and October 1998 were reviewed retrospectively. Of the 370 patients evaluated, 46 returned to theatre with compromised free flaps. The compromise was purely venous in origin in 37 of these cases, arterial in three and due to a combination of arterial and venous problems in six. Thirty-five of the flaps were successfully salvaged. On average, the clinical manifestation of the problem occurred 25.5 hours postoperatively. However, there was a significant time difference between flaps that were salvaged successfully and those that were not: in the salvaged group the compromise was identified 17.5 hours postsurgery compared with 51 hours for the unsuccessful group. The timing of the return to theatre following the identification of the compromise was a significant factor in the success rate: 71 minutes for those salvaged and 103 minutes for those not salvaged. It is recommended that flaps are monitored hourly for the first 72 postoperative hours and observations recorded on a chart.

Activation of protein kinase C stimulates the dephosphorylation of natriuretic peptide receptor-B at a single serine residue: a possible mechanism of heterologous desensitization.

The binding of atrial natriuretic peptide and C-type natriuretic peptide (CNP) to the guanylyl cyclase-linked natriuretic peptide receptors A and B (NPR-A and -B), respectively, stimulates increases in intracellular cGMP concentrations. The vasoactive peptides vasopressin, angiotensin II, and endothelin inhibit natriuretic peptide-dependent cGMP elevations by activating protein kinase C (PKC). Recently, we identified six in vivo phosphorylation sites for NPR-A and five sites for NPR-B and demonstrated that the phosphorylation of these sites is required for ligand-dependent receptor activation. Here, we show that phorbol 12-myristate 13-acetate, a direct activator of PKC, causes the dephosphorylation and desensitization of NPR-B. In contrast to the CNP-dependent desensitization process, which results in coordinate dephosphorylation of all five sites in the receptor, phorbol 12-myristate 13-acetate treatment causes the dephosphorylation of only one site, which we have identified as Ser(523). The conversion of this residue to alanine or glutamate did not reduce the amount of mature receptor protein as indicated by detergent-dependent guanylyl cyclase activities or Western blot analysis but completely blocked the ability of PKC to induce the dephosphorylation and desensitization of NPR-B. Thus, in contrast to previous reports suggesting that PKC directly phosphorylates and inhibits guanylyl cyclase-linked natriuretic peptide receptors, we show that PKC-dependent dephosphorylation of NPR-B at Ser(523) provides a possible molecular explanation for how pressor hormones inhibit CNP signaling.

Three years' experience of collaborative care pathways on a maxillofacial ward.

Collaborative care pathways (CCPs) provide a framework for multidisciplinary patient care. They provide guidelines and a mechanism for audit, and were first introduced at the Regional Unit, Walton Hospital, Liverpool, in November 1994. They have been designed for many surgical groups. Between August 1996 and 31 July 1997, 955 patients were admitted on to the nine established pathways: fractured mandible (n=213), fractured zygoma (n=117), minor oral surgery (n=244), abscess (n=18), examination under anaesthesia (n=73), nasal surgery (n=73), osteotomy (n=80), salivary (n=63), and temporomandibular joint (n=74). The purpose of this article is to report the introduction of CCP in a maxillofacial ward and give results from a one-year audit. CCP have proved to be an extremely useful tool and have several advantages over traditional documentation. They are more accurate, easily computerized, and facilitate audit. They promote the development of guidelines and standardized perioperative care, and this in turn facilitates training and raises standards of care.

Essential roles for the products of the napABCD genes, but not napFGH, in periplasmic nitrate reduction by Escherichia coli K-12.

The seven nap genes at minute 47 on the Escherichia coli K-12 chromosome encode a functional nitrate reductase located in the periplasm. The molybdoprotein, NapA, is known to be essential for nitrate reduction. We now demonstrate that the two c-type cytochromes, the periplasmic NapB and the membrane-associated NapC, as well as a fourth polypeptide, NapD, are also essential for nitrate reduction in the periplasm by physiological substrates such as glycerol, formate and glucose. None of the three iron-sulphur proteins, NapF, NapG or NapH, are essential, irrespective of whether the bacteria are grown anaerobically in the presence of nitrate or fumarate as a terminal electron acceptor, or by glucose fermentation. Mutation of napD resulted in the total loss of Methyl Viologen-dependent nitrate reductase activity of the molybdoprotein, NapA, consistent with an earlier suggestion by others that NapD might be required for post-translational modification of NapA.

A constitutively "phosphorylated" guanylyl cyclase-linked atrial natriuretic peptide receptor mutant is resistant to desensitization.

Dephosphorylation of the natriuretic peptide receptor-A (NPR-A) is hypothesized to mediate its desensitization in response to atrial natriuretic peptide (ANP) binding. Recently, we identified six phosphorylation sites within the kinase homology domain of NPR-A and determined that the conversion of these residues to alanine abolished the ability of the receptor to be phosphorylated or to be activated by ANP and ATP. In an attempt to generate a form of NPR-A that mimics a fully phosphorylated receptor but that is resistant to dephosphorylation, we engineered a receptor variant (NPR-A-6E) containing glutamate substitutions at all six phosphorylation sites. Consistent with the known ability of negatively charged glutamate residues to substitute functionally, in some cases, for phosphorylated residues, we found that NPR-A-6E was activated 10-fold by ANP and ATP. As determined by guanylyl cyclase assays, the hormone-stimulated activity of the wild-type receptor declined over time in membrane preparations in vitro, and this loss was blocked by the serine/threonine protein phosphatase inhibitor microcystin. In contrast, the activity of NPR-A-6E was more linear with time and was unaffected by microcystin. The nonhydrolyzable ATP analogue adenosine 5'-(beta,gamma-imino)-triphosphate was half as effective as ATP in stimulating the wild-type receptor but was equally as potent in stimulating NPR-A-6E, suggesting that ATP is required to keep the wild-type but not 6E variant phosphorylated. Finally, the desensitization of NPR-A-6E in whole cells was markedly blunted compared with that of the wild-type receptor, consistent with its inability to shed the negative charge from its kinase homology domain via dephosphorylation. These data provide the first direct test of the requirement for dephosphorylation in guanylyl cyclase desensitization and they indicate that it is an essential component of this process.