Bases para o estudo da genética de populações dos Hymenoptera em geral e dos Apinae sociais em particular

This paper deals with problems on population genetics in Hymenoptera and particularly in social Apidae. 1) The studies on populations of Hymenoptera were made according to the two basic types of reproduction: endogamy and panmixia. The populations of social Apinae have a mixed method of reproduction with higher percentage of panmixia and a lower of endogamy. This is shown by the following a) males can enter any hive in swarming time; b) males of Meliponini are expelled from hives which does not need them, and thus, are forced to look for some other place; c) Meliponini males were seen powdering themselves with pollen, thus becoming more acceptable in any other hive. The panmixia is not complete owing to the fact that the density of the breeding population as very low, even in the more frequent species as low as about 2 females and 160 males per reproductive area. We adopted as selection values (or survival indices) the expressions according to Brieger (1948,1950) which may be summarised as follows; a population: p2AA + ²pq Aa + q2aa became after selection: x p2AA + 2pq Aa + z q²aa. For alge-braics facilities Brieger divided the three selective values by y giving thus: x/y p2 AA + y/y 2 pq Aa + z/y q²aa. He called x/y of RA and z/y of Ra, that are survival or selective index, calculated in relation to the heterozygote. In our case all index were calculated in relation to the heterozygote, including the ones for haploid males; thus we have: RA surveval index of genotype AA Ra surveval index of genotype aa R'A surveval index of genotype A R'a surveval index of genotype a 1 surveval index of genotype Aa The index R'A ande R'a were equalized to RA and Ra, respectively, for facilities in the conclusions. 2) Panmitic populations of Hymenoptera, barring mutations, migrations and selection, should follow the Hardy-Weinberg law, thus all gens will be present in the population in the inicial frequency (see Graphifc 1). 3) Heterotic genes: If mutation for heterotic gene ( 1 > RA > Ra) occurs, an equilibrium will be reached in a population when: P = R A + Ra - 2R²a _____________ (9) 2(R A + Ra - R²A - R²a q = R A + Ra - 2R²A _____________ (10) 2(R A + Ra - R²A - R²a A heterotic gene in an hymenopteran population may be maintained without the aid of new mutation only if the survival index of the most viable mutant (RA) does not exced the limiting value given by the formula: R A = 1 + √1+Ra _________ 4 If RA has a value higher thah the one permitted by the formula, then only the more viable gene will remain present in the population (see Graphic 10). The only direct proof for heterotic genes in Hymenoptera was given by Mackensen and Roberts, who obtained offspring from Apis mellefera L. queens fertilized by their own sons. Such inbreeding resulted in a rapid loss of vigor the colony; inbred lines intercrossed gave a high hybrid vigor. Other fats correlated with the "heterosis" problem are; a) In a colony M. quadrifasciata Lep., which suffered severely from heat, the percentage of deths omong males was greater .than among females; b) Casteel and Phillips had shown that in their samples (Apis melifera L). the males had 7 times more abnormalities tian the workers (see Quadros IV to VIII); c) just after emerging the males have great variation, but the older ones show a variation equal to that of workers; d) The tongue lenght of males of Apis mellifera L., of Bombus rubicundus Smith (Quadro X), of Melipona marginata Lep. (Quadro XI), and of Melipona quadrifasciata Lep. Quadro IX, show greater variationthan that of workers of the respective species. If such variation were only caused by subviables genes a rapid increasse of homozigoty for the most viable alleles should be expected; then, these .wild populations, supposed to be in equilibrium, could .not show such variability among males. Thus we conclude that heterotic genes have a grat importance in these cases. 4) By means of mathematical models, we came to the conclusion tht isolating genes (Ra ^ Ra > 1), even in the case of mutations with more adaptability, have only the opor-tunity of survival when the population number is very low (thus the frequency of the gene in the breeding population will be large just after its appearence). A pair of such alleles can only remain present in a population when in border regions of two races or subspecies. For more details see Graphics 5 to 8. 5) Sex-limited genes affecting only females, are of great importance toHymenoptera, being subject to the same limits and formulas as diploid panmitic populations (see formulas 12 and 13). The following examples of these genes were given: a) caste-determining genes in the genus Melipona; b) genes permiting an easy response of females to differences in feeding in almost all social Hymenoptera; c) two genes, found in wild populations, one in Trigona (Plebéia) mosquito F. SMITH (quadro XII) and other in Melipona marginata marginata LEP. (Quadro XIII, colonies 76 and 56) showing sex-limited effects. Sex-limited genes affecting only males do not contribute to the plasticity or genie reserve in hymenopteran populations (see formula 14). 6) The factor time (life span) in Hymenoptera has a particular importance for heterotic genes. Supposing one year to be the time unit and a pair of heterotic genes with respective survival indice equal to RA = 0, 90 and Ra = 0,70 to be present; then if the life time of a population is either one or two years, only the more viable gene will remain present (see formula 11). If the species has a life time of three years, then both alleles will be maintained. Thus we conclude that in specis with long lif-time, the heterotic genes have more importance, and should be found more easily. 7) The colonies of social Hymenoptera behave as units in competition, thus in the studies of populations one must determine the survival index, of these units which may be subdivided in indice for egg-laying, for adaptive value of the queen, for working capacity of workers, etc. 8) A study of endogamic hymenopteran populations, reproduced by sister x brother mating (fig. 2), lead us to the following conclusions: a) without selection, a population, heterozygous for one pair of alleles, will consist after some generations (theoretically after an infinite number of generation) of females AA fecundated with males A and females aa fecundated with males a (see Quadro I). b) Even in endogamic population there is the theoretical possibility of the presence of heterotic genes, at equilibrium without the aid of new mutations (see Graphics 11 and 12), but the following! conditions must be satisfied: I - surveval index of both homozygotes (RA e Ra) should be below 0,75 (see Graphic 13); II - The most viable allele must riot exced the less viable one by more than is permited by the following formula (Pimentel Gomes 1950) (see Gra-fic 14) : 4 R5A + 8 Ra R4A - 4 Ra R³A (Ra - 1) R²A - - R²a (4 R²a + 4 Ra - 1) R A + 2 R³a < o Considering these two conditions, the existance of heterotic genes in endogamic populations of Hymenoptera \>ecames very improbable though not - impossible. 9) Genie mutation offects more hymenopteran than diploid populations. Thus we have for lethal genes in diploid populations: u = q2, and in Hymenoptera: u = s, being u the mutation ratio and s the frequency of the mutant in the male population. 10) Three factors, important to competition among species of Meliponini were analysed: flying capacity of workers, food gathering capacity of workers, egg-laying of the queen. In this connection we refer to the variability of the tongue lenght observed in colonies from several localites, to the method of transporting the pollen in the stomach, from some pots (Melliponi-ni storage alveolus) to others (e. g. in cases of pillage), and to the observation that the species with the most populous hives are almost always the most frequent ones also. 11) Several defensive ways used for Meliponini to avoid predation are cited, but special references are made upon the camouflage of both hive (fig. 5) and hive entrance (fig. 4) and on the mimetism (see list in page ). Also under the same heading we described the method of Lestrimelitta for pillage. 12) As mechanisms important for promoting genetic plasticity of hymenopteran species we cited: a) cytological variations and b) genie reserve. As to the former, duplications and numerical variations of chromosomes were studied. Diprion simile ATC was cited as example for polyploidy. Apis mellife-ra L. (n •= 16) also sugests polyploid origen since: a) The genus Melipona, which belongs to a" related tribe, presents in all species so far studied n = 9 chromosomes and b) there occurs formation of dyads in the firt spermatocyte division. It is su-gested that the origin of the sex-chromosome of Apis mellifera It. may be related to the possible origin of diplo-tetraploidy in this species. With regards to the genie reserve, several possible types of mutants were discussed. They were classified according to their survival indices; the heterotic and neutral mutants must be considered as more important for the genie reserve. 13) The mean radius from a mother to a daghter colony was estimated as 100 meters. Since the Meliponini hives swarm only once a year we may take 100 meters a year as the average dispersion of female Meliponini in ocordance to data obtained from Trigona (tetragonisca) jaty F. SMITH and Melipona marginata LEP., while other species may give different values. For males the flying distance was roughly estimated to be 10 times that for females. A review of the bibliography on Meliponini swarm was made (pg. 43 to 47) and new facts added. The population desity (breeding population) corresponds in may species of Meliponini to one male and one female per 10.000 square meters. Apparently the males are more frequent than the females, because there are sometimes many thousands, of males in a swarm; but for the genie frequency the individuals which have descendants are the ones computed. In the case of Apini and Meliponini, only one queen per hive and the males represented by. the spermatozoos in its spermateca are computed. In Meliponini only one male mate with the queen, while queens of Apis mellijera L. are fecundated by an average of about 1, 5 males. (Roberts, 1944). From the date cited, one clearly sees that, on the whole, populations of wild social bees (Meliponini) are so small that the Sewall Wright effect may become of great importance. In fact applying the Wright's formula: f = ( 1/aN♂ + 1/aN♀) (1 - 1/aN♂ + 1/aN♀) which measures the fixation and loss of genes per generation, we see that the fixation or loss of genes is of about 7% in the more frequent species, and rarer species about 11%. The variation in size, tergite color, background color, etc, of Melipona marginata Lep. is atributed to this genetic drift. A detail, important to the survival of Meliponini species, is the Constance of their breeding population. This Constance is due to the social organization, i. e., to the care given to the reproductive individuals (the queen with its sperm pack), to the way of swarming, to the food storage intended to control variations of feeding supply, etc. 14) Some species of the Meliponini are adapted to various ecological conditions and inhabit large geographical areas (e. g. T. (Tetragonisca jaty F. SMITH), and Trigona (Nanno-trigona testaceicornis LEP.) while others are limited to narrow regions with special ecological conditions (e. g. M. fuscata me-lanoventer SCHWARZ). Other species still, within the same geographical region, profit different ecological conditions, as do M. marginata LEP. and M. quadrifasciata LEP. The geographical distribution of Melipona quadrifasciata LEP. is different according to the subspecies: a) subsp anthidio-des LEP. (represented in Fig. 7 by black squares) inhabits a region fron the North of the S. Paulo State to Northeastern Brazil, ,b) subspecies quadrifasciata LEP., (marked in Fig. 7 with black triangles) accurs from the South of S. Paulo State to the middle of the State of Rio Grande do Sul (South Brazil). In the margined region between these two areas of distribution, hi-brid colonies were found (Fig. 7, white circles); they are shown with more details in fig. 8, while the zone of hybridization is roughly indicated in fig. 9 (gray zone). The subspecies quadrifasciata LEP., has 4 complete yellow bands on the abdominal tergites while anthidioides LEP. has interrupted ones. This character is determined by one or two genes and gives different adaptative properties to the subspecies. Figs. 10 shows certains meteorological isoclines which have aproximately the same configuration as the limits of the hybrid zone, suggesting different climatic adaptabilities for both genotypes. The exis-tance of a border zone between the areas of both subspecies, where were found a high frequency of hybrids, is explained as follows: being each subspecies adapted to a special climatic zone, we may suppose a poor adaptation of either one in the border region, which is also a region of intermediate climatic conditions. Thus, the hybrids, having a combination of the parent qualities, will be best adapted to the transition zone. Thus, the hybrids will become heterotic and an equilibrium will be reached with all genotypes present in the population in the border region.

Spectrophotometric determination of arsenic in soil samples using 2-(5-bromo-2-pyridylazo)-5-di-ethylaminophenol (Br-PADAP)

In this paper, a new, simple and sensitive method for arsenic determination in soil is proposed. This is based on the reduction of silver (I) and iron (III) ions by arsine followed by a complexation reaction of iron (II) with the spectrophotometric reagent Br-PADAP 2-(5-bromo-2-pyridylazo)-5-di-ethylaminophenol. Arsenic determination with a Sandell's sensitivity of 3.1 10-4 cm-2, linear range from 0.1 µg ml-1 to 2.0 µg ml-1 (r560 = 0.9995), molar absorptivity of 2.45 10(5) l mol-1 cm-1 and a concentration detection limit of 1.4 ng ml-1 (3s) were obtained using a 10 ml sample volume. Selectivity was increased with the use of EDTA as a masking agent. The proposed method was applied for arsenic determination in the presence of several ions amounts in digested soil samples. The results revealed that antimony (III), mercury (II), germanium (IV), platinum (IV) interferes at all analyzed proportions. The interferences can be easily removed by the use of EDTA. Precision and accuracy obtained were satisfactory with a R.S.D. < 5 %. Recovery of arsenic in soil samples varied from 95.55 to 102.70 % with a mean of 99.63 %. These results demonstrated that the proposed method is applicable for arsenic analysis in different soil samples.

Doença de depósito lisossomal induzida pelo consumo de Ipomoea verbascoidea (Convolvulaceae) em caprinos no semiárido de Pernambuco

O objetivo deste trabalho foi reproduzir a intoxicação por Ipomoea verbascoidea em caprinos e descrever os aspectos epidemiológicos, clínicos e histopatológicos da intoxicação espontânea por essa planta no Estado de Pernambuco. Para isso, realizou-se o acompanhamento da epidemiologia da doença em sete municípios do semiárido pernambucano. Três caprinos espontaneamente intoxicados foram examinados e, em seguida eutanasiados e necropsiados (Grupo I). Para reproduzir experimentalmente a doença, as folhas secas de I. verbascoidea contendo 0,02% de swainsonina, foram fornecidas na dose de 4g/kg (0,8mg de swainsonina/kg) a dois grupos de três animais. Os caprinos do Grupo II receberam a planta diariamente por 40 dias e foram eutanasiados no 41º dia de experimento. Os caprinos do Grupo III receberam a planta diariamente por 55 dias e foram eutanasiados no 120º dia de experimento. Outros três caprinos constituíram o grupo controle (Grupo IV). Nos grupos experimentais, as lesões encefálicas foram avaliadas por histopatologia e adicionalmente avaliaram-se as lesões cerebelares por morfometria, mediante mensuração da espessura da camada molecular, do número de neurônios de Purkinje e da área dos corpos celulares dessas células. Os principais sinais clínicos e lesões microscópicas foram semelhantes aos previamente reportados em animais intoxicados por plantas que contem swainsonina. Nos caprinos do GII e GIII, os primeiros sinais clínicos foram observados entre o 22º e 29º dia de experimento; clinicamente a doença desenvolvida por esses animais foi semelhante aos casos espontâneos. Nenhum dos caprinos do GIII se recuperou dos sinais neurológicos. Esse resultado evidencia que o consumo da planta por 26-28 dias após a observação dos primeiros sinais clínicos é suficiente para provocar lesões irreversíveis. Pela análise morfométrica, a camada molecular do cerebelo dos caprinos do Grupo I e III eram mais delgadas que às dos caprinos do grupo controle, e os neurônios de Purkinje estavam atróficos. Sugere-se que essas alterações sejam responsáveis pelo quadro clínico neurológico observado nos caprinos que deixam de ingerir a planta e apresentam seqüelas da intoxicação.

Troponina C na detecção imuno-histoquímica de alterações regressivas precoces no miocárdio de ovinos naturalmente intoxicados por Amaranthus spinosus (Amaranthaceae)

Resumo: Amaranthus spp. são plantas nefrotóxicas popularmente conhecidas como "caruru". Em casos de intoxicação por estas plantas, a principal alteração histopatológica está presente no rim, sob forma de nefrose tubular tóxica, porém em alguns casos pode haver alterações cardíacas. Alterações no eletrocardiograma, compatíveis com quadros de hipercalemia, foram descritas em suínos intoxicados por Amaranthus retroflexus e lesões como degeneração e necrose de miócitos cardíacos descritas em suínos intoxicados por A. caudatus e ovinos intoxicados por A. spinosus. Há dúvidas com relação às alterações cardíacas, que, na maioria dos casos, são incipientes, o que pode levar a erros de interpretação. Para a realização do trabalho foram utilizados blocos parafinados oriundos de um surto natural de intoxicação por A. spinosus no sudeste do Brasil. Esse estudo teve como objetivo detectar a presença de alterações regressivas incipientes no miocárdio de ovinos intoxicados por A. spinosus, através da utilização imuno-histoquímica do anticorpo anti-troponina C. Foram utilizados fragmentos de coração de 8 ovinos adultos e 2 fetos, intoxicados naturalmente por A. spinosus. Estes fragmentos foram submetidos à técnica de imuno-histoquímica com a utilização do anticorpo anti-troponina C. Pela avaliação imuno-histoquímica do coração dos oito ovinos adultos observaram-se diversos grupos de miócitos com diminuição significativa ou ausência de imunorreatividade para o anticorpo anti-troponina C; essas áreas correspondiam, em grande parte, aos mesmos grupos de miócitos que apresentavam, pela coloração de Hematoxilina e Eosina (H.E.) alterações que variavam de leve tumefação celular a aumento da eosinofilia, perda de estriação, lise celular e cariólise, ou mais raramente, acompanhadas de infiltrado inflamatório. Em quatro casos foi possível notar que diversos pequenos grupos de miócitos que tinham marcada diminuição de imunorreatividade, correspondiam a células com alterações imperceptíveis ou muito discretas no H.E. Nos corações dos dois fetos não houve áreas com perda ou diminuição de imunorreatividade para o anticorpo anti-troponina C. Os resultados da avaliação imuno-histoquímica confirmaram a presença tanto de alterações regressivas incipientes, quanto ratificaram a ocorrência de lesões necróticas já bem instaladas no miocárdio de ovinos intoxicados por Amaranthus spinosus. Verificou-se correspondência entre as áreas com ausência de imunorreatividade à troponina com as áreas de lesões mais marcadas no H.E.. Adicionalmente, a técnica foi capaz de detectar lesões muito precoces (locais do miocárdio sem sinais morfológicos de agressão), e demonstrou que as áreas agredidas/lesadas eram maiores que o evidenciado no HE. Esse estudo demonstra a necessidade de melhor se investigar a possível participação da hipercalemia e de outras alterações metabólicas presentes na lesão renal aguda, na gênese das lesões cardíacas de rápida instalação, ou seja, aquelas não correlacionadas às lesões vasculares determinadas pela uremia de longa duração.

Eficiência do 2,4-D aplicado isoladamente e em mistura com glyphosate no controle da trapoeraba

A trapoeraba (gênero Commelina) é planta daninha-problema em cafezais devido à sua capacidade de sobreviver nas mais diversas condições e tolerância ao herbicida glyphosate. O objetivo deste trabalho foi avaliar a eficiência do herbicida 2,4-D, em doses crescentes, aplicado isoladamente ou em mistura com o glyphosate, no controle de Commelina benghalensis e Commelina diffusa. Para isso, foi conduzido um experimento para cada espécie, em vasos, em casa de vegetação, no delineamento inteiramente casualizado, com cinco repetições. Foram comparados dez tratamentos, que consistiram da combinação de cinco doses de 2,4-D (0; 167,5; 335; 670; e 1.005 g ha-1) e duas doses de glyphosate (0 e 720 g ha¹). Avaliou-se a eficácia dos tratamentos pela porcentagem de controle, avaliada em cinco épocas, em relação à testemunha. Em C. benghalensis, 2,4-D proporcionou controle excelente (>91%) aos 33 DAT (dias após tratamento) a partir de 167,5 g ha-1 na presença de glyphosate e a partir de 335 g ha-1 na ausência de glyphosate. Em C. diffusa, 2,4-D proporcionou controle excelente aos 33 DAT a partir de 670 g ha-1, tanto na presença quanto na ausência de glyphosate. No entanto, somente a mistura de 2,4-D + glyphosate a 1.005 + 720 g ha-1 provocou 100% de controle desta espécie, verificando-se rebrota das plantas nos outros tratamentos. Nas condições dos experimentos, C. benghalensis mostrou-se mais suscetível que C. diffusa ao herbicida 2,4-D aplicado isoladamente ou em mistura com o glyphosate. Portanto, a identificação da espécie de trapoeraba presente na área a ser tratada e o conhecimento de sua biologia auxiliam na escolha do melhor produto e da dose ideal a ser aplicada, ou mesmo na escolha da mistura adequada, garantindo menor custo e melhor controle, com menores riscos para a cultura e o meio ambiente.

von Willebrand factor antigen levels in plasma of patients with malignant breast disease

von Willebrand factor (vWF) is a protein that mediates platelet adherence to the subendothelium during primary hemostasis. High plasma vWF concentrations have been reported in patients with various types of cancer, such as head and neck, laryngeal and prostatic cancer, probably representing an acute phase reactant. In the present study we determined the plasma levels of vWF antigen (vWF:Ag) by quantitative immunoelectrophoresis in 128 female patients with breast cancer as well as in 47 women with benign breast disease and in 27 healthy female controls. The levels of vWF:Ag were 170.7 ± 78 U/dl in patients with cancer, 148.4 ± 59 U/dl in patients with benign disease and 130.6 ± 45 U/dl in controls (P<0.005). We also detected a significant increase in the levels of vWF:Ag (P<0.0001) in patients with advanced stages of the disease (stage IV = 263.3 ± 113 U/dl, stage IIIB = 194.0 ± 44 U/dl) as compared to those with earlier stages of the disease (stage I = 155.3 ± 65 U/dl, stage IIA = 146.9 ± 75 U/dl). In conclusion, vWF levels were increased in plasma of patients with malignant breast disease, and these levels correlated with tumor progression.

Hepatitis C virus infection among Brazilian hemophiliacs: a virological, clinical and epidemiological study

We determined and analyzed risk factors of hepatitis C virus (HCV)-infected Brazilian hemophiliacs according to their virological, clinical and epidemiological characteristics. A cross-sectional and retrospective study of 469 hemophiliacs was carried out at a Brazilian blood center starting in October 1997. The prevalence of HCV infection, HCV genotypes and factors associated with HCV RNA detection was determined. The seroprevalence of anti-HCV antibodies (ELISA-3.0) was 44.6% (209/469). Virological, clinical and epidemiological assessments were completed for 162 positive patients. There were seven (4.3%) anti-HCV seroconversions between October 1992 and October 1997. During the same period, 40.8% of the positive anti-HCV hemophiliacs had abnormal alanine transaminase (ALT) levels. Plasma HCV RNA was detected by nested-RT-PCR in 116 patients (71.6%). RFLP analysis showed the following genotype distribution: HCV-1 in 98 hemophiliacs (84.5%), HCV-3 in ten (8.6%), HCV-4 in three (2.6%), HCV-2 in one (0.9%), and not typeable in four cases (3.4%). Univariate analysis indicated that older age (P = 0.017) and abnormal ALT levels (P = 0.010) were associated with HCV viremia, while the presence of inhibitor antibodies (P = 0.024) and HBsAg (P = 0.007) represented a protective factor against the presence of HCV RNA. These findings may contribute to a better understanding of the relationship between HCV infection and hemophilia.

Interferon-alpha receptor 1 mRNA expression in peripheral blood mononuclear cells is associated with response to interferon-alpha therapy of patients with chronic hepatitis C

Interferon (IFN)-alpha receptor mRNA expression in liver of patients with chronic hepatitis C has been shown to be a response to IFN-alpha therapy. The objective of the present study was to determine whether the expression of mRNA for subunit 1 of the IFN-alpha receptor (IFNAR1) in peripheral blood mononuclear cells (PBMC) is associated with the response to IFN-alpha in patients with chronic hepatitis C. Thirty patients with positive anti-HCV and HCV-RNA, and abnormal levels of alanine aminotransferase in serum were selected and treated with IFN-alpha2b for one year. Those with HBV or HIV infection, or using alcohol were not included. Thirteen discontinued the treatment and were not evaluated. The IFN-alpha response was monitored on the basis of alanine aminotransferase level and positivity for HCV-RNA in serum. IFNAR1-mRNA expression in PBMC was measured by reverse transcription-polymerase chain reaction before and during the first three months of therapy. The results are reported as IFNAR1-mRNA/ß-actin-mRNA ratio (mean ± SD). Before treatment, responder patients had significantly higher IFNAR1-mRNA expression in PBMC (0.67 ± 0.15; N = 5; P < 0.05) compared to non-responders (0.35 ± 0.17; N = 12) and controls (0.30 ± 0.16; N = 9). Moreover, IFNAR1-mRNA levels were significantly reduced after 3 months of treatment in responders, whereas there were no differences in IFNAR1 expression in non-responders during IFN-alpha therapy. Basal IFNAR1-mRNA expression was not correlated with the serum level of alanine and aspartate aminotransferases or the presence of cirrhosis. The present results suggest that IFNAR1-mRNA expression in PBMC is associated with IFN-alpha response to hepatitis C and may be useful for monitoring therapy in patients with chronic hepatitis C.

c-erbB-2 expression and nuclear pleomorphism in canine mammary tumors

The objective of the present investigation was to study the expression of c-erbB-2 and MIB-1 and try to associate them with morphological features of the cell such as nuclear pleomorphism, mitotic count and histological grade in a series of 70 canine mammary gland tumors, 22 of them benign and 48 malignant. Tumors were collected at the Veterinary Hospital of UFMG (Brazil) and the Veterinary Faculty of Porto University (Portugal). c-erbB-2 expression was determined according to the guidelines provided by the manufacturer of the HercepTest system and nuclear pleomorphism, mitotic count and histological grade according the Elston and Ellis grading system. The HercepTest is the FDA-approved in vitro diagnostic test marketed by Dako. It is a semi-quantitative immunohistochemical assay used to determine overexpression of HER2 protein (human epidermal growth factor receptor) in breast cancer tissue. MIB-1 expression was also evaluated in 28 malignant tumors. Seventeen (35.4%) of the malignant tumors were positive for c-erbB-2 expression, which was positively associated with nuclear pleomorphism (P < 0.0001), histological grade (P = 0.0017) and mitotic count (P < 0.05). Nuclear pleomorphism also showed a positive association with MIB-1 index (P < 0.0001). These results suggest that some of the biological and morphological characteristics of the tumor are associated in canine mammary gland tumors, as also reported for human breast cancer. It was also possible to show that the immunoexpression of c-erbB-2 can be a factor in mammary carcinogenesis. This fact opens the possibility of using anti-c-erbB-2 antibodies in the treatment of canine mammary tumors.

Validation of the Social Phobia and Anxiety Inventory for Children (SPAI-C) in a sample of Brazilian children

The purpose of the present study was to examine the factor structure and psychometric properties of the Social Phobia and Anxiety Inventory for Children (SPAI-C), an instrument developed in the United States and applied to a sample of Brazilian schoolchildren. The process included the translation of the original material from English into Portuguese by two bilingual psychiatrists and a back translation by a bilingual physician. Both the front and back translations were revised by a bilingual child psychiatrist. The study was performed using a cross-sectional design and the Portuguese version of the SPAI-C was applied to a sample of 1954 children enrolled in 3rd to 8th grade attending 2 private and 11 public schools. Eighty-one subjects were excluded due to an incomplete questionnaire and 2 children refused to participate. The final sample consisted of 1871 children, 938 girls (50.1%) and 933 boys (49.8%), ranging in age from 9 to 14 years. The majority of the students were Caucasian (89.0%) and the remainder were African-Brazilian (11.0%). The Pearson product-moment correlation showed that the two-week test-retest reliability coefficient was r = 0.780 and Cronbach's alpha was 0.946. The factor structure was almost similar to that reported in previous studies. The results regarding the internal consistency, the test-retest reliability and the factor structure were similar to the findings obtained in studies performed on English speaking children. The present study showed that the Portuguese language version of SPAI-C is a reliable and valid measure of social anxiety for Brazilian children.

Efeito de doses elevadas de vitamina D3 associada à suplementação de cálcio em modelo experimental de nefrolitíase

Introdução: Há poucos dados na literatura sobre a suplementação de vitamina D e cálcio e o desenvolvimento de cálculos renais. Objetivo: Avaliar o efeito de doses elevadas de vitamina D3 (V), com suplemento de cálcio (Ca) no desenvolvimento de litíase em modelo experimental. Métodos: Pastilhas foram inseridas na bexiga de ratos, que receberam V com ou sem Ca. Ratos foram divididos em seis grupos: 1. Sham; 2. Controle com pastilha, 3. Controle com V, 4. Pastilha + V, 5. Pastilha + Ca e 6. Pastilha + Ca + V. Resultados: Observou-se 50% e 17% de redução na formação de cálculos, respectivamente nos grupos 5 e 6 em comparação ao grupo 2 (p < 0,005). Não foram observadas hipercalcemia ou hipercalciúria em todos os grupos. Encontramos no grupo 6 (p = 0,03) uma redução significativa na calciúria. Conclusão: A administração de V associada com Ca diminuiu significantemente a formação de cálculos e reduziu significantemente a calciúria, sugerindo uma interferência benéfica na fisiopatologia litogênica.