Energy and mineral metabolism of peripartum mares and foals of the Quarter Horse breed.

BACKGROUND: Several studies have demonstrated variations in peripartum blood biochemical analytes used to monitor the health status of mares and their foals of different breeds. OBJECTIVES: We aimed to characterize the physiologic changes of peripartum energy and mineral metabolism in healthy Quarter Horse mares and their neonatal foals. METHODS: Blood samples were initially collected from 17 mares on days 60 and 30 prepartum, and from mares and their foals on the day of parturition (after colostrum ingestion), and on days 15, 30, and 60 postpartum. The serum concentrations of calcium, phosphorus, magnesium, total cholesterol (T-Cho), triglyceride, beta-hydroxybutyrate (BHBA), and nonesterified fatty acids (NEFA) were measured using commercial kits. Statistical differences were analyzed using one-way ANOVA followed by Tukey's test (parametric variables) or the Kruskal-Wallis test followed by Dunn's multiple comparison test (nonparametric variables). RESULTS: Fourteen of 17 healthy Quarter Horse mares and their neonatal foals remained in the study until termination. Serum BHBA, T-Cho, and calcium concentrations in mares showed significant differences during the observation period. Serum BHBA concentrations increased from 60 days prepartum to 60 days postpartum. Postpartum serum T-Cho and calcium concentrations significantly decreased until day-30 postpartum. In the foals, NEFA, T-Cho, calcium, and magnesium concentrations significantly differed between sampling times. In the foals, serum NEFA concentrations decreased from parturition until day-60 postpartum, while serum T-Cho, calcium, and magnesium concentrations were lowest at day-30 postpartum. CONCLUSIONS: The temporal changes observed in the blood biochemical analytes of this study could be used to improve the clinical evaluation of periparturient mare and neonatal foal Quarter Horses.

The role of motivation on physical activity and screen time behaviors among parent-adolescent dyads: The FLASHE study.

Behavioral theories inform the development of lifestyle interventions to address low participation in physical activity (PA); however, relatively little is known about the value of self-determination theory (SDT) for explaining screen time (ST) behaviors or in extending SDT into a dyadic context. Actor-partner (i.e., parent-adolescent) interdependence models (APIMs) allow for examination of these interpersonal relationships. The purpose of this study was to examine PA and ST among parent-adolescent dyads using the cross-sectional Family Life, Activity, Sun, Health, and Eating (FLASHE) Study. Parent-adolescent dyads provided responses to online surveys addressing PA (n = 1177 dyads) and ST (n = 1489 dyads) behaviors. We examined the influence of SDT-based constructs (perceived competence and motivation) on PA and ST behaviors. Structural equations were used to estimate APIMs in STATA 15.1. Full models provided a good fit to the data. For both PA and ST, perceived competence was more strongly associated with motivation among adolescents compared with parents (PA: ß = 0.72 vs. 0.58, ST: ß = 0.34 vs. 0.22, p's < 0.001). Parental motivation was associated with parental PA and both adolescent motivation for PA and ST (p's < 0.001). Parental motivation was not associated with adolescent ST-behavior. Adolescent motivation was only associated with parent motivation for PA. In the FLASHE study, SDT constructs extend acceptably to the dyadic setting, with PA models providing a slightly better fit to the data than ST models. Longitudinal studies that target perceived competence and the self-regulation of motivation in parents and their adolescents are a next logical step to understanding both PA and ST behaviors.

Ultrasonographic characterization of the organs in the middle third and caudal coelomic cavity of snakes in the Boidae Family / Caracterização ultrassonográfica dos órgãos do terço médio e caudal da cavidade celomática de serpentes da família Boidae

The aim of this study was the ultrasonographic evaluation of the organs in the middle third and caudal coelomic cavity of healthy snakes in the Boidae family. For such, 15 adult snakes from five species were evaluated: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi and Epicrates crassus. The animals were physically restrained for examination. After the application of acoustic gel over the scales the transducer was positioned in the frontal plane of the coelomic cavity. Microconvex and linear transducers at 10MHz were used. The ultrasound examination was conducted in the craniocaudal direction, with longitudinal and transversal sections to assess syntropy, echogenicity and echotexture of the organs. The liver was bilobulated, hyperechoic, homogeneous echotexture, hyperechoic capsule, with a hepatic lobe measuring 1.15±0.64cm wide, and the presence of a central hepatic vein, measuring 0.44±0.21cm in diameter. The gallbladder was characterized as an anechoic structure with echogenic walls and dimensions of 2.91±1.18cm x 1.38±0.84cm (length x width). The stomach showed a pleated and linear texture, echogenic, with hypoechoic walls measuring 0.3±0.07cm wide. The splenopancreas was a circular and echogenic structure, homogeneous echotexture and measuring 1.18±0.62cm in width and 1.56±0.88cm in length. The kidneys were characterized by elongated, lobulated, hypoechoic structures, an echogenic capsule and an echogenic central line, measuring 1.05±0.7cm in width. The testis were characterized by fusiform, hypoechoic structures, homogeneous echotexture, measuring 2.31±0.79cm in length and 0.6±0.23cm in width. Ovarian follicles were seen in different stages, 0.67±0.39cm wide and 0.73±0.38cm long. No differences were found between species of syntropy, echogenicity and echotexture of the organs of the coelomic cavity. The ultrasound examination proved to be a safe, non-invasive and efficient technique for characterizing the organs of the coelomic cavity of snakes.(AU)

O objetivo do estudo foi realizar a avaliação ultrassonográfica de órgãos do terço médio e caudal da cavidade celomática de serpentes hígidas da família Boidae. Para isto, foram avaliadas 15 serpentes adultas de cinco espécies: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi e Epicrates crassus. Os animais foram contidos fisicamente para a realização do exame. Após aplicação de gel acústico sobre as escamas, o transdutor foi posicionado no plano frontal da cavidade celomática. Foram utilizados transdutores microconvexo e linear em 10MHz. O exame ultrassonográfico foi conduzido no sentido craniocaudal, com cortes longitudinais e transversais para avaliar sintopia, ecogenicidade e ecotextura dos órgãos. O fígado se apresentou bilobulado, hiperecoico, ecotextura homogênea, cápsula hiperecoica, com lobo hepático medindo 1,15±0,64cm de largura, e presença de uma veia central hepática, medindo 0,44±0,21cm de diâmetro. A vesícula biliar foi caracterizada como uma estrutura anecoica de paredes ecogênicas e dimensões de 2,91±1,18cm x 1,38±0,84cm(comprimento x largura). O estômago apresentou-se com o aspecto pregueado e linear, ecogênico, com paredes hipoecoicas medindo 0,3±0,07cm de largura. O esplenopâncreasse apresentou como uma estrutura circular e ecogênica, ecotextura homogênea e medindo de 1,18±0,62cm de largura e 1,56±0,88cm de comprimento. Os rins se caracterizaram por estruturas alongadas, lobuladas, hipoecoicas, cápsula ecogênica e uma linha central ecogênica, medindo 1,05±0,7cm de largura. Os testículos foram caracterizados por estruturas fusiformes, hipoecoicas, ecotextura homogênea, medindo 2,31±0,79cm de comprimento e 0,6±0,23cm de largura. Foram visualizados folículos ovarianos em diferentes estágios, de 0,67±0,39cm de largura e 0,73±0,38cm de comprimento. Não foram encontradas diferenças entre espécies de sintopia, ecogenicidade e ecotextura dos órgãos da cavidade celomática. O exame ultrassonográfico demonstrou-se uma técnica segura, não invasiva e eficiente para caracterização dos órgãos da cavidade celomática de serpentes.(AU)

Ultrasonographic characterization of the organs in the middle third and caudal coelomic cavity of snakes in the Boidae Family

ABSTRACT: The aim of this study was the ultrasonographic evaluation of the organs in the middle third and caudal coelomic cavity of healthy snakes in the Boidae family. For such, 15 adult snakes from five species were evaluated: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi and Epicrates crassus. The animals were physically restrained for examination. After the application of acoustic gel over the scales the transducer was positioned in the frontal plane of the coelomic cavity. Microconvex and linear transducers at 10MHz were used. The ultrasound examination was conducted in the craniocaudal direction, with longitudinal and transversal sections to assess syntropy, echogenicity and echotexture of the organs. The liver was bilobulated, hyperechoic, homogeneous echotexture, hyperechoic capsule, with a hepatic lobe measuring 1.15±0.64cm wide, and the presence of a central hepatic vein, measuring 0.44±0.21cm in diameter. The gallbladder was characterized as an anechoic structure with echogenic walls and dimensions of 2.91±1.18cm x 1.38±0.84cm (length x width). The stomach showed a pleated and linear texture, echogenic, with hypoechoic walls measuring 0.3±0.07cm wide. The splenopancreas was a circular and echogenic structure, homogeneous echotexture and measuring 1.18±0.62cm in width and 1.56±0.88cm in length. The kidneys were characterized by elongated, lobulated, hypoechoic structures, an echogenic capsule and an echogenic central line, measuring 1.05±0.7cm in width. The testis were characterized by fusiform, hypoechoic structures, homogeneous echotexture, measuring 2.31±0.79cm in length and 0.6±0.23cm in width. Ovarian follicles were seen in different stages, 0.67±0.39cm wide and 0.73±0.38cm long. No differences were found between species of syntropy, echogenicity and echotexture of the organs of the coelomic cavity. The ultrasound examination proved to be a safe, non-invasive and efficient technique for characterizing the organs of the coelomic cavity of snakes.

RESUMO: O objetivo do estudo foi realizar a avaliação ultrassonográfica de órgãos do terço médio e caudal da cavidade celomática de serpentes hígidas da família Boidae. Para isto, foram avaliadas 15 serpentes adultas de cinco espécies: Boa constrictor amarali, Boa constrictor constrictor, Eunectes murinus, Epicrates assisi e Epicrates crassus. Os animais foram contidos fisicamente para a realização do exame. Após aplicação de gel acústico sobre as escamas, o transdutor foi posicionado no plano frontal da cavidade celomática. Foram utilizados transdutores microconvexo e linear em 10MHz. O exame ultrassonográfico foi conduzido no sentido craniocaudal, com cortes longitudinais e transversais para avaliar sintopia, ecogenicidade e ecotextura dos órgãos. O fígado se apresentou bilobulado, hiperecoico, ecotextura homogênea, cápsula hiperecoica, com lobo hepático medindo 1,15±0,64cm de largura, e presença de uma veia central hepática, medindo 0,44±0,21cm de diâmetro. A vesícula biliar foi caracterizada como uma estrutura anecoica de paredes ecogênicas e dimensões de 2,91±1,18cm x 1,38±0,84cm(comprimento x largura). O estômago apresentou-se com o aspecto pregueado e linear, ecogênico, com paredes hipoecoicas medindo 0,3±0,07cm de largura. O esplenopâncreasse apresentou como uma estrutura circular e ecogênica, ecotextura homogênea e medindo de 1,18±0,62cm de largura e 1,56±0,88cm de comprimento. Os rins se caracterizaram por estruturas alongadas, lobuladas, hipoecoicas, cápsula ecogênica e uma linha central ecogênica, medindo 1,05±0,7cm de largura. Os testículos foram caracterizados por estruturas fusiformes, hipoecoicas, ecotextura homogênea, medindo 2,31±0,79cm de comprimento e 0,6±0,23cm de largura. Foram visualizados folículos ovarianos em diferentes estágios, de 0,67±0,39cm de largura e 0,73±0,38cm de comprimento. Não foram encontradas diferenças entre espécies de sintopia, ecogenicidade e ecotextura dos órgãos da cavidade celomática. O exame ultrassonográfico demonstrou-se uma técnica segura, não invasiva e eficiente para caracterização dos órgãos da cavidade celomática de serpentes.

Heavy Metal Accumulation in the Intestinal Tapeworm Proteocephalus macrophallus Infecting the Butterfly Peacock Bass (Cichla ocellaris), from Southeastern Brazil.

Here we evaluate the potential of heavy metal accumulation of Proteocephalus macrophallus parasitizing the Butterfly Peacock Bass (Cichla ocellaris). A total of 19 fish specimens were collected. From the hosts, samples of intestine, liver, muscle, and parasites were taken. Heavy metal concentrations (Al, As, Ba, Cd, Cr, Fe, Hg, Mg, Mn, Ni, Pb, Ti, and Zn) were obtained using Atomic Absorption Spectrometry. All analyzed elements was found in higher concentrations in the parasites comparing to its host tissues. The bioconcentration factors were higher in the intestine, varying between 5.91 (Ti) to 8.00 (Ba), followed by the muscle, 1.88 (Mg) to 6.39 (Zn), and liver, 1.67 (Al) to 2.02 (Ba). These results show that at the infection site heavy metal concentrations are reduced, since the elements are absorbed directly from the intestinal wall by the parasites. In general, P. macrophallus presents a reasonable capacity of metal accumulation comparing to its hosts.

Contracaecum sp. parasitizing Acestrorhynchus lacustris as a bioindicator for metal pollution in the Batalha River, southeast Brazil.

Pollution in aquatic ecosystems due to negative human activities remains a problem in both freshwater and marine environments and is an ongoing subject of research. Several studies have shown that some fish parasites can be used as a tool for biomonitoring because they demonstrate higher metal accumulation capacity compared to their host tissues. However, compared to acanthocephalans, information regarding the absorption mechanisms and accumulation rates in nematodes is relatively limited. Here, we evaluated the potential of larvae Contracaecum sp. (L3) as indicators of contamination by metals by analyzing thirteen element concentrations: magnesium (Mg), aluminum (Al), titanium (Ti), chromium (Cr), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), cadmium (Cd), barium (Ba), and lead (Pb) in the parasites and host Acestrorhynchus lacustris, using inductively coupled plasma mass spectrometry. Twelve of the thirteen analyzed elements were detected in at least 2-fold higher concentrations (e.g. Ni) and were up to approximately 50-fold higher (e.g. Pb) in parasites than in host tissues, including elements known for their high toxicity (As, Cd, Pb) and those considered to be essential (Cu and Zn). Our results suggest that Contracaecum sp. larvae can be used as bioindicators of metal contamination because even in early stages of development, numerous essential and non-essential elements were accumulated, making this system a useful tool for monitoring polluted environments.

Global TLR2 and 4 deficiency in mice impacts bone resorption, inflammatory markers and atherosclerosis to polymicrobial infection.

Toll-like-receptors (TLRs) play a significant role in the generation of a specific innate immune response against invading pathogens. TLR2 and TLR4 signaling contributes to infection-induced inflammation in periodontal disease (PD) and atherosclerosis. Observational studies point towards a relationship between PD and atherosclerosis, but the role of TLR2 and TLR4 in the recognition of multiple oral pathogens and their modulation of host response leading to atherosclerosis are not clear. We evaluated the role of TLR2 and TLR4 signaling in the induction of both PD and atherosclerosis in TLR2-/- and TLR4-/- mice to polymicrobial infection with periodontal pathogens Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum. Polybacterial infections have established gingival colonization in TLR2-/- and TLR4-/- mice and induction of a pathogen-specific immunoglobulin G immune response. But TLR deficiency dampened accelerated alveolar bone resorption and intrabony defects, indicating a central role in infection-induced PD. Periodontal bacteria disseminated from gingival tissue to the heart and aorta through intravascular dissemination; however, there was no increase in atherosclerosis progression in the aortic arch. Polybacterial infection does not alter levels of serum risk factors such as oxidized low-density lipoprotein, nitric oxide, and lipid fractions in both mice. Polymicrobial-infected TLR2-/- mice demonstrated significant levels (P < 0.05 to P < 0.01) of T helper type 2 [transforming growth factor-ß1 , macrophage inflammatory protein-3α, interleukin-13 (IL-13)] and T helper type 17 (IL-17, IL-21, IL-22, IL-23) splenic T-cell cytokine responses. Increased heat-shock protein expression, hspa1a for Hsp 70, was observed for both TLR2-/- and TLR4-/- mice. This study supports a role for TLR2 and TLR4 in PD and atherosclerosis, corroborating an intricate association between two inflammatory diseases.

Increased atherogenesis during Streptococcus mutans infection in ApoE-null mice.

Streptococcus mutans, a dental caries pathogen, also causes endocarditis and is detected in atheroscelerotic plaque. We investigated the potential for an invasive strain of S. mutans, OMZ175, to accelerate plaque growth in apolipoprotein E deficient (ApoE(null)) mice without and with balloon angioplasty (BA) injury, a model of restenosis. ApoE(null) mice were divided into 4 groups (N = 10), 2 with and 2 without BA. One each of the BA and non-BA groups was infected with S. mutans (Sm). S. mutans DNA, plaque area, inflammatory cell invasion, and Toll-like receptor (TLR) expression were measured at 6-20 weeks post-infection. S. mutans genomic DNA was detected in the aorta, liver, spleen, and heart. Plaque growth was significantly increased in infected mice with BA (Sm+BA) vs. those in the non-infected groups (p < 0.03). Plaque size was increased after infection without BA (Sm), but did not reach significance. Aortic specimens from both S. mutans and Sm+BA groups displayed increased numbers of macrophages, and TLR4 expression was increased in BA mice. In conclusion, S. mutans infection accelerated plaque growth, macrophage invasion, and TLR4 expression after angioplasty. S. mutans may also be associated with atherosclerotic plaque growth in non-injured arteries.

Bax expression in untreated breast cancer: an immunocytometric study of 255 cases.

BACKGROUND: Bax is one of the main effectors of apoptosis in breast cancer. However, in contrast with the antiapoptotic protein Bcl-2, which has been extensively studied in this tumor, there are relatively few clinical studies on the biological role of Bax in breast cancer. MATERIALS AND METHODS: The expression of the apoptosis-related Bax gene was studied in a series of 255 previously untreated breast cancers by means of immuno-flow cytometry. Additionally, and by the same method, the expression of the Bcl-2, VEGF and Nup88 genes were also studied. As variables of the study for the final statistical analysis, the histological variety of the tumors, histological and nuclear grade, the expression of hormone receptors, p53, Ki-67 or c-erb-B2, axillary node invasion, tumor size and DNA-ploidy were also included. RESULTS: The expression of the proapoptotic Bax protein was significantly associated with the expression of Nup88 (p<0.0001), VEGF (p=0.0014) and Bcl-2 (p=0.0063), all measured by the same method. An inverse correlation with c-erb-B2 expression, which almost attained statistical significance (p=0.058) was also registered. CONCLUSION: This study adds evidence to the little explored link between apoptosis and angiogenesis. Furthermore, it discloses a previously unreported relationship between Bax and Nup88 expression.

The small variant of the apoptosis-associated X-chromosome RBM10 gene is co-expressed with caspase-3 in breast cancer.

BACKGROUND: There are very few studies on the final triggers of apoptosis, the caspases, in breast cancer. MATERIALS AND METHODS: Caspase-3 expression was studied by means of reverse transcript polymerase chain reaction (RT-PCR) in a series of 108 previously untreated patients with breast cancer. Expression levels were correlated with those obtained using the same technique of the apoptosis-associated X-chromosome genes RBMX, RBM3, RBM10 small and RBM10 large variant; Bcl-2 and Bax; the angiogenesis-associated genes VEGF and CD105 (endoglin); hMAM and Nup88. The correlation with the expression of hormone receptors, c-erb-B2, mutant p53 and Ki-67, all measured by means of immunohistochemistry, was also studied, as well as that with standard clinical parameters such as histological type, tumor size, axillary metastasis and DNA-ploidy. RESULTS: The only statistically significant correlations observed between caspase-3 mRNA expression and the parameters tested were a direct one with both the Bax (p = 0.007) and the small variant of the X-chromosome RBM10 gene (p = 0.018), and an inverse one with the angiogenesis-associated CD105 (endoglin) gene (p = 0.044). CONCLUSION: These results indicate that very few genes are involved in the last steps of the apoptotic cascade in breast cancer, among them one of the X-chromosome RBM family. They also support the relatively unexplored link between apoptosis and angiogenesis.

Clasificación molecular simplificada del carcinoma ductal infiltrante unifocal pT1: identificación de subgrupos biológicamente diferenciados / No disponible

Antecedentes: Trabajos recientes, utilizando la tecnologíade “microarrays” han resultado en una nueva clasificaciónpronóstica del cáncer de mama basada en el perfil genético delos tumores. Las diferencias en la supervivencia de las pacientespertenecientes a los diferentes subgrupos que estableceesta nueva clasificación son altamente significativas. Sin embargo,ninguno de los citados estudios ha estratificado los tumorespor histologías o por tamaños. En el presente trabajo,hemos reclasificado nuestros cánceres de mama más precoces(pT1), pertenecientes exclusivamente al tipo ductal infiltrante,de acuerdo con una versión simplificada de la clasificación moleculararriba aludida, propuesta por sus propios autores.Material y métodos: Estudiamos 346 cánceres de mamaductales infiltrantes, unifocales, pT1. De ellos, 251 (72,5%) seencuadraron en el grupo luminal A (RE+, RP+, c-erb-B2 -); 45(13%) fueron triple-negativos, 20 (5,8%) expresaron al mismotiempo receptores hormonales y c-erb-B2 (tipo luminal B), y30 (8,7%) expresaron c-erb-B2 en ausencia de receptores hormonales(tipo HER2).Resultados: Durante el análisis estadístico, resultó evidenteque los tumores tipo luminal B y los HER2 presentaban idénticas,o casi idénticas correlaciones con los factores clínicos ymoleculares estudiados, y fueron finalente encuadrados en unsolo grupo de tumores con expresión de c-erb-B2, sin teneren cuenta la coexpresión o no de receptores hormonales.Comparando los tumores del grupo luminal A con el resto,los primeros mostraron una correlación significativamente inversacon un grado histológico y nuclear 3 (r = - 0,15, p =0,004 y r = - 0,40, p < 0,0001, respectivamente), con la expresiónde p53 (r = - 0,45, p < 0,0001) y un índice de KI67superior al 20% (r = - 0,42, p < 0,0001)...(AU)

Background: Recent studies using microarray technologyhave resulted in a new prognostic classification of breast cancer,based on the genetic profile of the tumors. The differencesin survival between the women belonging to the differentsubgroups established by this new classification are highlysignificant. However, none of the cited studies has stratifiedthe tumors by histology or size. In the present paper, we havereclassified our early breast cancers (pT1) belonging exclusivelyto the ductal infiltrating variety, according to a simplifiedversion of the above cited molecular classification, as proposedby their authors.Materials and methods: We studied 346 unifocal, pT1ductal infiltrating carcinomas. Of them, 251 (72.5%) wereclassified as luminal A type (ER+, PR+, c-erb-B2 -); 45 (13%)were triple-negative, 20 (5.8%) expressed at the same timehormonal receptors and c-erb-B2 (luminal B type), and 30(8.7%) expressed c-erb-B2, with absent hormone receptor expression(HER2 type).Results: During the statistical workup, it became evidentthat luminal B and HER2-type tumors showed identical, or almostidentical correlations with the studied clinical and molecularparameters, and were finally regrouped into a single categoryof c-erb-B2-expressing tumors, regardless of theirhormone receptor status. Comparing the tumors of the luminal A group with therest, the former showed a significant inverse correlation withhistologic and nuclear grade 3 (r = - 0.15, p = 0.004 and r = -0.40, p < 0.0001, respectively), with p53 expression (r = -0.45, p < 0.0001), as well as with a Ki67 labelling index>20% (r = - 0.42, p < 0.0001)...(AU)

Viral anti-inflammatory reagents: the potential for treatment of arthritic and vasculitic disorders.

Inflammatory and immune responses are inherent in the development of progressive arthritic or vasculitic disorders. Arthritis is frequently associated with accelerated forms of vasculitis; atherosclerosis being one form of accelerated vasculitis that blocks blood flow causing heart attacks and strokes. The arterial supply is central to maintaining normal articular function and acts as a conduit for inflammatory (innate) and immune (antigen dependent) cell trafficking in joints. The vasculature in some cases can become inflamed in the disease process. While treatment of severely debilitating arthritic disorders has improved, some current treatments are limited to reducing symptoms while others act as disease modifying drugs (DMARDs), but may have limited success. Many current treatments also have reported adverse side effects. Vasculitic disorders are similarly debilitating with high associated morbidity and mortality and current therapy for these disorders is only partially successful. Immune-modifying agents, which alter vascular inflammation, thus have potential for application in rheumatologic diseases. Viral immune modulating proteins reduce early arterial inflammatory responses with associated reductions in atherosclerotic plaque development and transplant rejection in a wide range of animal models. A clinical trial utilizing one such viral reagent, a secreted myxomaviral serpin, is currently in progress, assessing treatment of acute coronary syndrome, a vascular syndrome with marked up-regulation of systemic inflammatory responses. In this review we examine viral anti-inflammatory proteins as potential therapeutic reagents for arthritic and vasculopathic disorders.

The S-phase fraction of the aneuploid cell subpopulation is the biologically relevant one in aneuploid breast cancers.

BACKGROUND: In the case of DNA-aneuploid tumors there are no clear guidelines as to which S-phase fraction is the more relevant one: that corresponding to either the diploid or the aneuploid population, or rather an average of both. MATERIALS AND METHODS: We studied 280 breast cancer specimens from previously untreated patients. Histologically, 231 were ductal infiltrating carcinomas, 30 lobular infiltrating carcinomas and 19 corresponded to other, less frequent varieties. Postsurgically, 164 cases (58.6%) were classified as T1, 87 (31.1%) as T2 and 7 as T3. The remaining 22 cases were multifocal, diffuse tumors. Flow cytometry was performed on fresh tumor tissue, and immunohistochemistry for hormone receptors, Ki67, c-erb-B2 and p53 on paraffin-embedded material. RESULTS: In diploid tumors, a high S-phase (above the 75th percentile) correlated significantly with Ki67 expression > or =20% (p<0.0001). In aneuploid tumors, however, this was only the case for the aneuploid fraction of tumor cells (p< 0.0001). A high S-phase of diploid tumors correlated directly and significantly with a high histologic grade (p=0.04), a high nuclear grade (p=0.01), tumor size (p=0.0008), and inversely with estrogen (p<0.0001) and progesterone (p<0.0001) receptor expression. In aneuploid tumors, the aneuploid tumor fraction showed a direct and significant correlation with a high histologic grade (p=0.005), a high nuclear grade (p=0.001), mutant p53 expression (p=0.0009), and inversely with estrogen (p<0.0001) and progesterone (p=0.0001) receptor expression. A high S-phase of the diploid cell fraction of aneuploid tumors, on the other hand, just showed an inverse correlation with high nuclear grade of the tumors (p=0.02), and none whatsoever with all other tested parameters.

Extirpación de carcinomas no palpables de mama mediante el sistema ABBI. Estudio anatomopatológico de la glándula mamaria circumlesional / Extirpation of nonpalpable breast carcinomas using the Advanced Breast Biopsy Instrumentation (ABBI) system. Pathological study of the circumlesional mammary gland

No disponible

Elevated mammaglobin (h-MAM) expression in breast cancer is associated with clinical and biological features defining a less aggressive tumour phenotype.

BACKGROUND: Mammaglobin (h-MAM) is expressed mainly by breast epithelial cells, and this feature has been used to detect circulating breast cancer cells and occult metastases in sentinel axillary lymph nodes of breast cancer patients. However, the biological role of mammaglobin is completely unknown. METHODS: We studied 128 fresh-frozen breast cancer specimens by means of reverse transcriptase-polymerase chain reaction and quantified their h-MAM mRNA expression. This was then correlated with histological and nuclear grade, oestrogen and progesterone receptor expression, c-erb-B2 and mutant p53 expression, as well as with cellular proliferation measured by means of the Ki67 labelling index, DNA ploidy and S-phase, and finally with the presence or not of invaded axillary nodes in the mastectomy specimen. RESULTS: In the univariate analysis, high h-MAM expression (above the median for the whole group) correlated significantly (P < 0.05) with oestrogen and progesterone receptor expression, diploid DNA content, low Ki67 labelling index, low nuclear grade and almost significantly (P = 0.058) with the absence of axillary nodal invasion in the mastectomy specimen. In a final, multivariate model, only progesterone receptor expression, diploid DNA content and absence of nodal invasion were found to be independently associated with high h-MAM expression. CONCLUSION: All of the features associated with mammaglobin expression reflect, without exception, a less aggressive tumour phenotype. Further studies are needed to clarify whether this is attributable to h-MAM expression itself, or to another mechanism of which mammaglobin expression forms part.

Immunofluorometric study of Bcl-2 and Bax expression in clinical fresh tumor samples from breast cancer patients.

BACKGROUND: The balance between the expression of the antiapoptotic gene Bcl-2 and the proapoptotic gene Bax is considered a good indicator of the apoptotic activity of tumor cells. Bcl-2 and Bax expression seem also to individually play a prognostic role in breast cancer. Our aim was to study the expression of both genes in fresh breast cancer samples, and to correlate the obtained results with other available clinical and biological parameters of the tumors. MATERIALS AND METHODS: Fresh tumor specimens from 86 breast cancer patients were studied by means of immunofluorocytometry for the expression of the apoptosis-associated Bcl-2 and Bax genes. Additionally, DNA-ploidy was also measured. Paraffin blocks corresponding to the same tumors were used for immunohistochemistry, to study the expression of hormone receptors (ER and PR), p53, c-erb-B2 and the Ki67 labelling index. Fourteen patients had been treated with four cycles of induction chemotherapy (cyclo-phosphamide, adriamycin and 5-fluorouracil), and separate statistical analyses were carried out both for the whole group, and for the 62 patients not having received any treatment whatsoever, in order to exclude any potential influence of the chemotherapeutic treatment on the expression of the studied antigens. RESULTS: Bcl-2 expression correlated significantly with estrogen (p = 0.002) and progesterone (p = 0.012) receptor expression, as well as with c-erb-B2 (p = 0.045) expression in chemotherapy-naïve tumors, the correlation being completely lost if treated tumors were added to the study group. A high apoptotic index (Bcl2/Bax < 0.5) correlated significantly with progesterone receptor expression (p = 0.037) and c-erb-B2 expression (p = 0.018), and this correlation was maintained, whether previously treated tumors were included into the study or not (p = 0.038 and p = 0.027, respectively). Bax expression did not correlate with any other clinical or biological parameters of the tumors, including Bcl-2 expression. CONCLUSION: Bcl-2 and Bax-expression can be easily determined in clinical breast cancer specimens by means of immunofluorocytometry. Bcl-2-expression significantly correlates with hormone-receptor- and c-erb-B2-expression exclusively in previously untreated tumors. This, however, seems only to be the case when considering Bcl-2 expression in isolation, since a high apoptotic index, which considers the ratio of Bcl-2 versus Bax expression in the same tumor, seems not to be affected by the previous use of chemotherapy.

Accumulation of uPA-PAI-1 complexes inside the tumour cells is associated with axillary nodal invasion in progesterone-receptor-positive early breast cancer.

Both urokinase-like plasminogen activator (uPA) and its inhibitor plasminogen activator inhibitor (PAI-1), as well as uPA-PAI-1 complexes, have been identified as important prognostic factors in breast cancer. We have recently reported that the latter are identifiable inside breast cancer cells by means of immunohistochemistry. Using this technique, we have studied a series of 212 early (pT1) unifocal breast cancers and have correlated the expression of uPA-PAI-1 complexes, together with other clinical and biological features (histologic variety, histologic and nuclear grade, hormone receptors, Ki67 labelling index, c-erb-B2-, p53- and CD44std-expression) with or without the occurrence of axillary node invasion. In a logistic regression model, looking for associations with axillary metastasis, we found a statistically significant interaction between the presence of uPA-PAI-1 complexes and progesterone receptor positivity (P=0.04). A final model showed that the presence of uPA-PAI-1 complexes was a determinant factor for axillary metastasis among women carrying tumours expressing progesterone receptors. In these cases, the presence of uPA-PAI-1 complexes carried with it a nearly 14-fold risk of axillary node invasion (P=0.009). These results may indicate that small, hormone-receptor-positive breast cancers (with a theoretical good prognosis) may carry an elevated risk of nodal involvement if accumulation of uPA-PAI-1 complexes is shown inside their tumour cells by means of immunohistochemistry.

PAI-1 promoter polymorphism modulates uPA-PAI complex accumulation by breast cancer cells.

OBJECTIVES: The uPA-PAI system has been shown to play a role in the development of a more aggressive tumor phenotype. The PAI-1 promoter 4G/5G polymorphism, furthermore, regulates free plasma PAI-1 levels in patients with myocardial infarction. Our aim was to verify if the different polymorphisms in the PAI-1 promoter are also associated with alterations in the intracellular accumulation of uPA-PAI complexes in human breast cancer. METHODS: Accumulation of uPA-PAI complexes inside the tumor cells was determined by means of immunohistochemistry, as previously described by our own group, and two extremely different sets of tumors were chosen, one of them with strong uPA-PAI complex reactivity inside more than 50% of tumor cells, the other with no demonstrable reactivity at all. Finally, the 4G/5G polymorphism of the PAI-1 promoter was studied in all of them by means of DNA extraction, PCR amplification of the PAI promoter sequence, and restriction polymorphism typing. RESULTS: Absence of intracellular uPA-PAI complex accumulation was significantly associated with the prevalence of the 4G allele and, conversely, the presence of uPA-PAI complexes inside the tumor cells was significantly associated with 5G/5G homozygosity (logistic regression, p = 0.0128). Furthermore, none of the 7 5G/5G homozygous tumors showed histological grade 3, as did 6/21 tumors in the group where the 4G allele was present. In spite of the low case number, this association of the 5G/5G polymorphism with a less aggressive phenotype almost reached statistical significance (Spearman's correlation test, p = 0.118). CONCLUSIONS: The 4G/5G polymorphism of the PAI-1 promoter seems indeed to be associated with different rates of uPA-PAI complex internalization by breast cancer cells. Complex accumulation inside the tumor cells is significantly related to 5G/5G homozygosity, and this shows a trend towards an association with a less aggressive, better-differentiated tumor phenotype.

Flow cytometry vs. Ki67 labelling index in breast cancer: a prospective evaluation of 181 cases.

BACKGROUND: Excessive proliferation is one of the first steps in oncogenic activation and one of the most important biological features defining the aggressiveness of tumors. Quantifying the proportion of tumor cells in S-phase by means of flow cytometry has shown, in the past, to be useful for defining high-risk subgroups in breast cancer. Several antigens closely associated with proliferation are also detectable by means of immunohistochemistry, offering in theory an easy to perform and cheap alternative to flow cytometry for measuring proliferation. To test this hypothesis, we compared both methods prospectively in a series of breast cancers. MATERIALS AND METHODS: We studied the proliferation rate of 181 breast cancers (152 ductal infiltrating, 17 lobular infiltrating, 12 other histological varieties), operated upon at our institution, by means of flow cytometry and the Ki67 labelling index, using the MIBI antibody. Ploidy (expressed as DNA content or DNA-index), S-phase fraction and the Ki67 labelling index were the variables of the study. The S-phase fraction was considered separately for diploid and aneuploid tumors, following the 1992 Maine Consensus guidelines and was judged abnormally elevated if higher than the 75th percentile for each group. The Ki67 labelling index was expressed as percent positive tumor cells, positive cells being those showing specific nuclear staining. RESULTS: DNA-ploidy and the Ki67 labelling index could be evaluated in all tumors. Of the total, 96 (53%) were diploid and 85 (47%) aneuploid. S-phase fraction could be measured in 172 out of the 181 tumors (95%). The 75th percentile cut-offs for diploid and aneuploid tumors were 9.9% and 15.8%, respectively. We found a significant correlation beween rising DNA content and increasing Ki67 index (r = 0.18; p = 0.022), as well as between the percentage of cells in S-phase of the whole tumor population and Ki67 (r = 0.22; p = 0.0055). A Ki67 cut-off of 50% or higher identified most aneuploid tumors, or a small group of diploid ones with a high S-phase fraction (specificity = 96.7%; positive predictive value 92.5%), however at the price of a very low sensitivity (62.6%). This was due to the presence of many aneuploid tumors with a low S-phase fraction. CONCLUSION: The Ki67 labelling index and S-phase fraction are significantly correlated. However, flow cytometry provides additional indirect information on tumor aggressiveness associated with DNA-ploidy. Further studies are needed to determine whether Ki67 alone is sufficient as a routinely applicable method.

Relationship of obesity and specimen weight to complications in reduction mammaplasty.

Obesity and specimen weight have both been associated with a higher incidence of complications for patients undergoing reduction mammaplasty. However, obesity has been arbitrarily and inconsistently defined, and the degree of obesity has not been considered in these previous studies. Because insurance companies are increasingly demanding weight loss before authorizing surgery, the relationship of obesity and breast size to complications is of great importance. Upon critical review of the literature, a number of fundamental questions remain unanswered. If complications are more frequent in the obese patient, are these complications directly proportional to the degree of obesity? Also, if the patient is required to lose weight before surgery, is weight loss effective in reducing complication rates? In an attempt to clarify these issues, 395 patients who underwent reduction mammaplasty over a 10-year period were reviewed retrospectively. Patients were arbitrarily divided into five groups in which, depending on their degree of relative obesity, they were classified as less than 5 percent, 5 to 10 percent, 10 to 15 percent, 15 to 20 percent, or greater than 20 percent above average body weight. To evaluate the relationship of specimen weight to complications, patients were also arbitrarily divided into five groups according to weight of the breast reduction specimen, which was classified as less than 300 g, 300 to 600 g, 600 to 900 g, 900 to 1200 g, and greater than 1200 g reduction per breast. Complications were then divided into local and systemic and major and minor. When bilateral reductions alone were analyzed (n = 267), there was a statistically significant increase in complication rate in the obese (p = 0.01). However, when the obese population was further subdivided according to their degree of obesity (less than 5 percent, 5 to 10 percent, 10 to 15 percent, 15 to 20 percent, and greater than 20 percent above average body weight), no further correlation was found. However, the relationship between specimen weight per breast and complications was much stronger with a direct correlation existing between increasing specimen weight and the incidence of complications. Although this study has shown that patients who are average body weight have fewer complications than obese patients after breast reduction surgery, it has not shown an increasing incidence of complication with increasing degrees of obesity. The implications of these findings and their relationship for denying patients surgery on the basis of weight alone are discussed in detail.