Angiogenic and lymphangiogenic microvessel density in breast carcinoma: correlation with clinicopathologic parameters and VEGF-family gene expression.

Angiogenesis and lymphangiogenesis are essential for breast cancer progression and are regulated by vascular endothelial growth factors (VEGF). To determine clinical and molecular correlates of these processes, we measured blood and lymphatic vascular microvessel density in 29 invasive carcinomas (22 ductal, six lobular, one papillary), using the vascular marker CD31 and the novel lymphatic marker D2-40. Microvessel density was assessed microscopically and by image cytometry, and was compared with tumor histology, grade, stage, lymph node metastasis, hormone receptors, HER2/neu status, and expression of VEGF, VEGF-C and VEGF-D by immunohistochemistry or quantitative RT-PCR. Strong correlation was observed between visual and image cytometric microvessel density using D2-40 but not CD31 (P=0.016 and 0.1521, respectively). Image cytometric CD31 microvessel density correlated with tumor size, grade, stage and lymph node metastasis (P=0.0001, 0.0107, 0.0035 and 0.0395, respectively). D2-40 microvessel density correlated with tumor stage (P=0.0123 by image cytometry) and lymph node metastasis (P=0.0558 by microscopy). Immunohistochemical VEGF signal in peritumoral blood vessels correlated with image cytometric CD31 and D2-40 microvessel density (P=0.022 and 0.0012, respectively), consistent with the role of VEGF in blood and lymphatic vascular growth. Intratumoral VEGF-C and VEGF-D expression by quantitative RT-PCR correlated with D2-40 (P=0.0291 by image cytometry) but not with CD31 microvessel density, which could suggest a selective role of VEGF-C and VEGF-D in lymphangiogenesis. CD31 and D2-40 microvessel density correlated significantly with several prognostic factors, including lymph node metastasis. Thus, measurements of angiogenesis and lymphangiogenesis may have utility for breast cancer pathology, particularly for estimation of metastatic risk.     

Polyamines in Trichomonas vaginalis

Trichomonas vaginalis was grown in a modified Bushby's medium and putrescine, spermidine and spermine levels were determined in extracts from 24- and 48-h cultures and also in the culture media. All three polyamines were present in T. vaginalis extracts; the putrescine level and putrescine/spermidine ratio were much higher than those reported for other protozoa or for mammalian tissues. There were no significant differences between 24-h and 48-h amine levels per mg protein in these extracts, but amine levels per cell were higher at 24 than at 48 h. The spent culture media had a much higher putrescine content than corresponding uninoculated media and it was concluded that T. vaginalis secreted putrescine into the culture medium.     

Differential effects of Th1, monocyte/macrophage and Th2 cytokine mixtures on early gene expression for glial and neural-related molecules in central nervous system mixed glial cell cultures: neurotrophins, growth factors and structural proteins

Background  

In multiple sclerosis, inflammatory cells are found in both active and chronic lesions, and it is increasingly clear that cytokines are involved directly and indirectly in both formation and inhibition of lesions. We propose that cytokine mixtures typical of Th1 or Th2 lymphocytes, or monocyte/macrophages each induce unique molecular changes in glial cells.     

Rapid intraoperative immunohistochemical evaluation of sentinel lymph nodes for metastatic breast carcinoma.

OBJECTIVE: Sentinel lymph node (SLN) biopsy is an integral part of the surgical management of patients with breast cancer. Rapid immunohistochemistry (RIHC) has the potential to increase detection of metastatic carcinoma at the time of frozen section consultation. The authors assessed the accuracy and turnaround time of a newly developed RIHC method for pancytokeratin (RIHC-CK). METHODS: Sixty-six SLNs from 32 patients with breast carcinoma were examined for metastasis using the Zymed Sentinel Lymph Node Rapid IHC Kit. Intraoperative frozen sections (6 mum) of the SLNs were incubated with Zymed anti-pan-cytokeratin/HRP conjugate, diaminobenzidine (DAB), and stained with hematoxylin. Slides were ready within 8 minutes and were interpreted as positive or negative for metastatic carcinoma. Results were compared with previous intraoperative touch preparations, frozen sections, hematoxylin and eosin (Perm H&E), and AEl/3-immunostained permanent sections (Perm CK). RESULTS: Fourteen lymph nodes (19%) in 13 patients tested positive for metastatic carcinoma in Perm H&E, the gold standard. RIHC-CK had the highest sensitivity (92%) of the intraoperative tests, compared with touch preparations (64%) and frozen sections (80%). RIHC-CK showed 94% accuracy, compared with 96% (frozen section) and 93% (touch preparation). The RIHC technique took 8 minutes and was easy to perform and interpret. CONCLUSIONS: Zymed RIHC is a sensitive method for detecting breast cancer metastases in SLNs. The speed, accuracy, and ease of interpretation of the test allow for recognition of micrometastases (<2 mm) that might otherwise be undetectable by current methods of intraoperative evaluation. The prognostic significance and effect on surgical management of micrometastases in SLNs have yet to be determined.     

Effects of health information in youth on adult physical activity: 20‐Year study results from the Amsterdam growth and health longitudinal study

In the Amsterdam Growth and Health Longitudinal Study (AGAHLS), a group of apparently healthy males and females (n = 200) were interviewed about their physical activities on eight separate occasions over a period of 20 years between 13 and 33 years of age (multi‐measured group: MM). Information about their health was given based on their personally measured lifestyle (activity, diet, smoking) and biological risk characteristics for chronic diseases (medical check‐ups). A comparable group of boys and girls (n = 200) was only measured on two occasions (bi‐measured group: BM): at 13 and 33 years. Physical activity was estimated with a structured interview. Total physical activity and sports activity were estimated in three intensity levels (light, moderate, and heavy). It was hypothesized that the eight repeated medical check‐ups with health information in the MM group would result in a healthier lifestyle with respect to the determinants and levels of habitual physical activity compared to the BM group. Contrary to the hypothesis, males and females in the BM group showed a significantly higher increase or a lower decrease in physical activities compared to the MM group. This negative effect on the physical activity pattern at 33 years in the MM group may have been caused by more underreporting of physical activities than in the BM group. In conclusion, there does not appear to be a significant effect of long‐term (multi‐measured) health information with medical check‐ups during adolescence and young adulthood on level of physical activity in males and females at 33 years of age. Am. J. Hum. Biol. 14:448–456, 2002. © 2002 Wiley‐Liss, Inc.     

Longitudinal relationships between nutritional status, body composition, and physical fitness in rural children of South Africa: The Ellisras longitudinal study.

The objective of this study was to investigate the development and tracking of nutritional status, body composition and physical fitness, and the longitudinal relationship of changes in nutritional status, and body composition with changes in physical fitness over a 1-year period of follow-up. Studied were 380 boys and 322 girls aged 7-14 years from the Ellisras Longitudinal Study. Boys and girls were divided into two groups of pre-adolescence (<11 years) and adolescence (>10 years). High tracking coefficients (>8) were found for nutritional status, body mass index, and fat-free mass, while low tracking coefficients (<4) were found for the sum of skinfolds, fat mass, arm muscle area, and central fat distribution. Moderate and low tracking coefficients were found for the physical fitness items. Longitudinal regression analyses showed that physical fitness performances that require a high energy flux over a short period of time are affected by muscle wasting, whereas having a low body weight appears to be important for a good performance on other fitness items in these malnourished rural South African children.     

Extensive immune-mediated hippocampal damage in mice surviving infection with neuroadapted Sindbis virus

Viral infections of the central nervous system and immune responses to these infections cause a variety of neurological diseases. Infection of weanling mice with Sindbis virus causes acute nonfatal encephalomyelitis followed by clearance of infectious virus, but persistence of viral RNA. Infection with a neuroadapted strain of Sindbis virus (NSV) causes fatal encephalomyelitis, but passive transfer of immune serum after infection protects from fatal disease and infectious virus is cleared. To determine whether persistent NSV RNA is associated with neurological damage, we examined the brains of recovered mice and found progressive loss of the hippocampal gyrus, adjacent white matter, and deep cerebral cortex associated with mononuclear cell infiltration. Mice deficient in CD4(+) T cells showed less tissue loss, while mice lacking CD8(+) T cells showed lesions comparable to those in immunocompetent mice. Mice deficient in both CD4(+) and CD8(+) T cells developed severe tissue loss similar to immunocompetent mice and this was associated with extensive infiltration of macrophages. The number of CD4(+) cells and macrophage/microglial cells, but not CD8(+) cells, infiltrating the hippocampal gyrus was correlated with the number of terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling positive pyramidal neurons. These results suggest that CD4(+) T cells can promote progressive neuronal death and tissue injury, despite clearance of infectious virus.     

Immunocytochemical localization of dopamine-beta-hydroxylase in neurons of the human brain stem

The distribution of catecholaminergic cells in the human pons and medulla was illustrated by immunocytochemistry using a polyclonal antibody directed against the catecholamine synthetic enzyme, dopamine-beta-hydroxylase. The antibody specifically recognizes dopamine-beta-hydroxylase in putative adrenergic and noradrenergic neurons. The adrenergic and noradrenergic neurons are found in the brain stem from caudal levels of the medulla through the caudalmost levels of the midbrain. Large numbers of dopamine-beta-hydroxylase-positive cells were observed in cell groups of the medulla. Additionally, dopamine-beta-hydroxylase-positive cells were concentrated in the nucleus locus coeruleus and the nucleus subcoeruleus in the pons. These studies confirm that immunocytochemical localization of dopamine-beta-hydroxylase can be used to identify noradrenergic and adrenergic neurons and their terminal varicosities in the pons and medulla in routine autopsy material. These studies have also illustrated that the distribution of dopamine-beta-hydroxylase in the adult human brain is comparable to the distribution in other species.