Impact of Genetics on Low Bone Mass in Adults

Low bone mass in adults is a major risk factor for low‐impact fractures and is considered of complex origin because of interaction of environmental and genetic factors, each with modest effect. The objective was to assess the relative impact of genetics and environment and quantify the risk in relatives of osteopenic individuals. We studied 440 Icelandic nuclear families with 869 first‐degree relatives of both sexes. Index cases (male or female) had BMD in the lumbar spine or hip >1.5 SD less than sex‐matched controls. Heritability of BMD was estimated by maximum likelihood method, and variance component analysis was used to partition the genetic and environmental effects. Relative risk of low BMD (< ?1 SD) in first‐degree relatives was estimated, and heritable decrement in BMD was calculated compared with controls. Heritability was estimated as 0.61–0.66. Relative risk among first‐degree relatives was 2.28, and the yield of screening was as high as 36%. The genetic influence was consistent with one or a few genes with considerable effect in addition to multiple genes each with a small effect. The genetic deficit in BMD was already present before 35 yr of age and equaled bone loss during 8–30 yr after menopause. We confirmed that genetics are more important than environment to low bone mass in adults. Our results are consistent with a few underlying genes with considerable effect. The prevalence among first‐degree relatives of both sexes is common, suggesting that screening them should be cost effective and informative to elucidate the underlying genetics.     

Interstitial Fluid Pressure in Human Melanoma Xenografts Relationship to fractional tumor water content, tumor size, and tumor volume-doubling time

The interstitial fluid pressure (IFP) has been shown to be elevated in malignant tissue, but the possibility that IFP might be related to other pathophysiological parameters of the tissue has not been fully explored. The purpose of the study here reported was to measure the IFP in human melanoma xenografts and to search for possible correlations between tumor IFP and fractional tumor water content, tumor wet weight, or tumor volume-doubling time. Tumors of four melanoma lines (A-07, D-12, R-18, U-25), grown orthotopically in BALB/c-nu/nu mice, were included in the study. Tumor IFP, measured by using the wick-in-needle technique, ranged from 2 to 10 mm Hg (D-12), from 2 to 15 mm Hg (A-07 and U-25), and from 2 to 30 mm Hg (R-18). Statistically significant correlations between tumor IFP on the one hand and fractional tumor water content, tumor wet weight, or tumor volume-doubling time on the other were not found, whether the tumor lines were analyzed individually or together. These observations suggest that simple general relationships between the IFP and the other pathophysiological parameters measured here, might not exist in tumors.     

Species variations in distribution of S100 in retina. Demonstration with a monoclonal antibody and a polyclonal antiserum

The S100 protein has been found consistently in glial cells both in the central nervous system (CNS) and peripheral nervous system (PNS). However, in the retina we find substantial species variation in the distribution of this protein. Immunohistochemically, in the human retina we do not find any S100. In the rabbit retina it is present both in Müller cells and in astrocytes and in the chicken retina it is in neurons. This demonstrates how misleading it can be to use the distribution of a protein in one species to generalize about the distribution of the same protein in other species. It is also clear that even though immunohistochemical staining for the S100 protein could be used to study pathologic conditions that involve Müller cells in guinea pigs, hamster, rat, and rabbit retina it is going to be of limited value in investigations of the same conditions in the human eye.     

Distribution of the neuronal specific protein, 14-3-2, in central nervous system lesions of tuberous sclerosis

Summary The distribution of a neuronal specific enolase (14-3-2) in the central nervous system (CNS) lesions of tuberous sclerosis (TS) was examined using antiserum to 14-3-2 and the peroxidase antiperoxidase (PAP) method of Sternberger. In cortical tubers all the giant cells had intense cytoplasmic staining. Only occasional cells in the subependymal nodules were stained. All cells in the subependymal giant cell tumors were intensely stained. This indicates that the cortical giant cells and the giant cell subependymal tumors are of neuronal rather than astrocytic origin.     

The inhibition of clotting complexes of the extrinsic coagulation cascade by the phospholipase A2 isoenzymes from Naja nigricollis venom

Phospholipase A2 (PLA2) isoenzymes from Naja nigricollis venom exhibit anticoagulant activity with varying potencies. To determine which complexes in the extrinsic coagulation cascade are inhibited by these PLA2 enzymes, we examined their effects on the coagulation of bovine plasma initiated by the addition of thromboplastin, Russell's viper venom (RVV) or thrombin. The weakly anticoagulant PLA2 enzymes, CM-I and CM-II, prolonged clotting initiated by thromboplastin, but not that initiated by RVV or thrombin. The strongly anticoagulant enzyme, CM-IV, prolonged clotting initiated by both thromboplastin and RVV, but not clotting initiated by thrombin. To confirm the differences in their inhibitory properties, we examined the effect of these PLA2 enzymes on reconstituted extrinsic tenase and prothrombinase complexes. The weakly anticoagulant enzymes inhibited the tenase complex, but did not inhibit the prothrombinase complex, whereas the strongly anticoagulant enzyme inhibited both complexes. Thus the enzymes showed distinct differences in their inhibition patterns in the extrinsic coagulation cascade. Their dissimilarity in inhibition of the two phospholipid dependent activation steps probably reflects the difference in phospholipid requirements and/or mechanism of inhibition between the two complexes. Inhibition of successive amplification steps in the extrinsic coagulation cascade by CM-IV is consistent with its potency as a strongly anticoagulant PLA2.     

Neovascularization of the iris: an experimental model in cats

Neovascularization of the iris was induced in cats by removing the vitreous and lens and creating a rhegmatogenous retinal detachment. The presence of new blood vessels on the anterior surface of the iris was verified from the second month onward by slit lamp examination, as well as by light microscopy six to twelve months after the operation. Control eyes undergoing vitrectomy and lensectomy, but without retinal detachment, did not develop rubeosis iridis. This model may allow investigation into causes and therapy of rubeosis iridis.     

Vascular proliferation is important for clinical progress of endometrial cancer

Angiogenesis is essential for tumor growth, invasion, and metastatic spread. Whereas microvessel density (MVD) has been widely used as a measure of tumor-associated angiogenesis, we now wanted to examine the significance of other angiogenic markers, especially vascular proliferation (by Ki-67/factor VIII staining) and the degree of pericyte coverage [by alpha-smooth muscle actin (alpha-SMA)/factor VIII staining], in a large and population-based series of endometrial carcinoma with complete follow-up. Due to limited information on the role of lymphangiogenesis in these tumors, lymphatic vessel density (LVD) by LYVE-1 staining was also determined, as well as selected angiogenic factors [vascular endothelial growth factor (VEGF)-A, VEGF-C, VEGF-D and basic fibroblast growth factor (bFGF)], which could possibly be related to vascular proliferation and lymphangiogenesis. The information on angiogenic phenotype was related to clinicopathologic features and disease progress. Median vascular proliferation, as estimated by vascular proliferation index (VPI), was 3.9% and high VPI was associated with features of aggressive tumors and decreased survival. The prognostic effect of VPI was superior to that of MVD. Presence of pericyte coverage, as estimated by the alpha-SMA index (SMAI), was 35% and low SMAI was significantly associated with vascular invasion by tumor cells and impaired prognosis. Peritumoral lymphatic vessels (LVD-pt) were found in 39.5% of the cases and high LVD-pt was significantly associated with aggressive tumor features and decreased survival. In multivariate survival analysis, only the extent of vascular proliferation had independent prognostic effect, in addition to well-known clinicopathologic factors, whereas MVD did not have significant prognostic value. In conclusion, our study indicates that vascular proliferation is a meaningful variable in assessing the angiogenic phenotype of endometrial carcinoma.     

Familial risk of colon and rectal cancer in Iceland: Evidence for different etiologic factors?

The aim of this study was to characterize the familial risk of colon and rectal cancer using 2 population-based registries in Iceland, the Icelandic Cancer Registry and a genealogy database. The standardized incidence ratio (SIR) was used to estimate the risk among relatives of colorectal cancer index cases diagnosed in Iceland over a 46-year period (1955-2000). The 2,770 colorectal cancer patients had 23,272 first-degree relatives. Among first-degree relatives, there was an increased risk of both colon (SIR 1.47, 95% confidence interval (CI) 1.34-1.62) and rectal cancer (SIR 1.24, 95% CI 1.04-1.47). An increased risk of colon cancer was observed among siblings of colon cancer patients (SIR 2.03, 95% CI 1.76-2.33), whereas no such increase was observed for parents or offspring. Furthermore, the risk of rectal cancer was only increased among brothers (SIR 2.46 95% CI 1.46-3.89) of rectal cancer patients and not among their sisters (SIR 1.0 95% CI 0.40-2.06). The added risk of colon cancer among first-degree relatives was independent of site of colon cancer in the proband. Our results confirm that family history of colorectal cancer is a risk factor for the disease. However, family history has a different association with colon cancer than with rectal cancer, suggesting that the 2 cancer types may have different etiologic factors. Our results have implications for colon and rectal cancer screening programs.     

Loss of p63 and cytokeratin 5/6 expression is associated with more aggressive tumors in endometrial carcinoma patients

p63 and cytokeratin (CK) 5/6 are markers of basal and squamous differentiation in several normal epithelia and human tumors and are also suggested to be markers of progenitor or stem cells in certain stratified epithelia. In endometrial carcinoma, there is very limited information about the expression pattern of p63 or CK5/6 and no prognostic information. The aim of our study was to examine whether the expression of these markers was associated with a certain tumor phenotype in terms of other biomarkers, clinicopathologic characteristics and patient prognosis. Immunohistochemical expression of p63 and CK5/6 was examined using tissue microarrays (TMAs) in a large population-based series of 276 endometrial carcinomas with long and complete follow-up. Selected cases of normal and hyperplastic endometrium were examined for comparison (n = 15). Absence of p63 expression (70%) was significantly associated with nonendometrioid carcinomas, high histologic grade (FIGO), higher mitotic count and tumor cell proliferation by Ki-67, microsatellite instability (MSI) and loss of hMSH6 expression. A tendency toward reduced patient survival was also seen (p = 0.098). Presence of CK5/6 expression was more frequent in endometrioid tumors with squamous differentiation, while loss of CK5/6 expression (54%) was significantly associated with high FIGO stage, reduced beta-catenin expression, MSI and reduced patient survival (p = 0.0001); the latter was also found within the endometrioid subgroup (p = 0.0004). Multivariate survival analysis revealed that loss of CK5/6 expression had an independent prognostic impact in addition to well-known prognostic variables. Expression of both markers was increased in simple hyperplasia compared with normal endometrium. In complex hyperplasia, p63 expression was also increased, whereas CK5/6 was positive in areas with squamous differentiation only. Thus, loss of p63 or CK5/6 was associated with features of aggressive tumors, and lack of CK5/6 was significantly associated with reduced survival in multivariate analysis.     

Generation of monoclonal antibodies recognizing neuronal elements in formalin-fixed paraffin-embedded human tissue

We used formalin-fixed human spinal cord and dorsal root ganglia as immunogens to generate monoclonal antibodies (mAb) which immunohistochemically react with neurons in formalin-fixed human tissue sections. Three of the mAb recognized all neuronal populations studied, including those in spinal cord, dorsal root ganglia, cerebellum, and cerebrum. A fourth mAb recognized neurons within spinal cord, dorsal root ganglia and dentate nucleus of cerebellum but not those in cerebrum or cerebellar hemispheres. This mAb, unlike the other three, did not recognize murine neurons. These data indicate the feasibility of generating mAb suitable for analysis of human pathological material in its most readily available form, formalin-fixed paraffin-embedded tissue.