Loss of heterozygosity in tuberous sclerosis hamartomas

We have previously described in tuberous sclerosis (TSC) hamartomas the phenomenon of loss of heterozygosity (LOH) for DNA markers in the region of both the TSC2 gene on chromosome 16p13.3 and the TSC1 gene on 9q34. We now describe the spectrum of LOH in 51 TSC hamartomas from 34 cases of TSC. DNA was extracted from leucocytes or normal paraffin embedded tissue, and from frozen paraffin embedded hamartoma tissue from the same patient. The samples were analysed for 11 markers spanning the TSC1 locus and nine markers spanning the TSC2 locus. Twenty-one of 51 hamartomas showed LOH (41%). There was significantly more LOH on 16p13.3, with 16 hamartomas showing LOH around TSC2, and five in the vicinity of TSC1. No hamartoma showed LOH for markers around both loci. All the areas of LOH on chromosome 9 were large, but the smallest region of overlap lay between the markers D9S149 and D9S114, providing independent evidence for the localisation of the TSC1 gene. These data show that LOH is a common finding in a wide range of hamartomas, affecting the same TSC locus in different lesions from the same patient but not affecting both loci. These data support the hypothesis that both the TSC genes act as tumour suppressors and that the manifestations of TSC in patients with germline TSC mutations rise from "second hit" somatic mutations inactivating the remaining normal copy of the TSC gene.     

Measurement of activin in biological fluids by radioimmunoassay, utilizing dissociating agents to remove the interference of follistatin

BACKGROUND: The aim of this morphological investigation was to obtain more information about the structural and cellular mechanisms of interalveolar pore formation in postnatal lung development. Assuming that alveolar pore formation is related to the general thinning of interalveolar walls observed in the postnatal period, we have focused our attention on the topographical relationship between epithelial cells and connective tissue in the septum. Thereby we tried to formulate a uniform concept of pore formation. METHODS: After fixation with glutaraldehyde and osmiumtetroxide, tissue blocks of rat lungs aged 44 days were embedded in Epon. Serial sections were obtained in order to analyse precisely pores and supposed sites of pore formation (type II cells and thin spots in transsections of interalveolar walls). RESULTS: We made the following observations: there are pores with or without type II cells in the neighbourhood, and "pre-pores" with either fully transseptal granular pneumocytes, or thin spots in the interalveolar wall consisting of one or two layers of type I cell epithelium or of type II and type I cells without intervening connective tissue. CONCLUSIONS: From these findings we deduce that there is a general principle of interalveolar pore formation which consists in the formation of transseptal interepithelial cell contacts (i.e., between cells of type II and type I or type I and type I), promoted by the thinning of interalveolar walls in the stage of microvascular maturation. Within the zone of contact the cells thin out and give way to form an interalveolar opening.     

Cell surface chondroitin sulfate proteoglycans in tumor cell adhesion, motility and invasion

Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6-thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG.     

Nonsanction seat belt law enforcement: a modern day tale of two cities

This paper describes the implementation and evaluation of a nonsanction seat belt law enforcement program in two experimental communities of contrasting size in a state with a mandatory belt law. The main ingredients of the program were seat belt "salutes," public information and education, and limited use of inexpensive economic incentives. Driver shoulder belt use data collected before, during, and after the experimental programs, compared to similar data collected in a comparison community, showed the approach to be effective. While standard seat belt enforcement activities without incentives have been shown to be effective, many police departments, especially in smaller communities, are reluctant to make wholesale increases in seat belt citations. Although requiring some additional level of manpower and resources, a nonsanction approach to seat belt law enforcement can provide an alternate way of increasing belt use in these communities.     

Pseudo-tandem stenosis of the internal carotid artery

Two cases of pseudo-tandem stenosis of the internal carotid artery are presented. Proximal stenosis was produced by a large, focal atherosclerotic plaque. The severe intracranial "stenosis" was thought to be an angiographic artifact. Repeat cerebral angiography after carotid endarterectomy revealed normal intracranial arteries. The recognition of this entity is important because a decision to operate is based largely upon angiographic findings.     

Value of bone marrow biopsy in the diagnosis of amyloidosis

Amyloid involvement of the bone marrow is not commonly diagnosed before necropsy. Paraffin sections of the trephine bone biopsy specimen are superior to marrow aspiration cell smears for the antemortem diagnosis. Thirteen cases of amyloidosis were diagnosed from the bone biopsy specimen during a ten-year period. Amyloid was detected in only two of the corresponding aspirates. Three morphologic patterns of marrow involvement were found: vascular, focal extravascular/perivascular, and diffuse. Five (38%) of the cases were associated with multiple myeloma. An abnormal immunoglobulin was detected in the serum or urine or both in ten of 11 cases when determined. Although the bone marrow may not be the best site for the diagnosis of amyloidosis, it should not be neglected and marrow biopsies taken for other diagnostic reasons may "incidentally" reveal amyloid. Amyloidosis should be included in the list of non-primary hematologic conditions of the bone marrow in which the trephine biopsy may prove useful for diagnosis.     

Fungal products. Part XVII. Microbiological hydroxylation of gibberellin A9 and its methyl ester

Angiotensin II (A II) and analogues were tested for their ability to restore electrical and mechanical activity to cardiac muscle preparations in which the fast Na+ channels had been inactivated by partial depolarization (22-27 mM K+) or by tetrodotoxin (TTX). The partially depolarized or TTX-blocked preparations were chosen because under these conditions electrical and mechanical responses are primarily Ca2+ -dependent. In depolarized rabbit right atria, A II restored spontaneous mechanical and electrical activity (measured by both intracellular and extracellular recording techniques). The frequency of action potential discharge was concentration-dependent; the threshold concentration of A II was 10(-10) M, the ED50 was 8 X 10(-9) M, and the maximum effect was observed at 5 X 10(-8) M. In contrast, depolarized guinea pig atria were insensitive to A II, Sar1-angiotensin II, and des-Asp1-angiotensin II, even at concentrations as high as 10(-5) M. Rabbit papillary muscle (TTX-blocked), embryonic (18-day) chick heart (partially depolarized) and chick heart reaggregates (TTX-blocked) responded similarly to rabbit atria in that A II (9.6 X 10(-7) M) restored both electrical and mechanical activity. We found that in these preparations the action of A II was unaffected by propranolol (5.0 X 10(-6) M to 5.0 X 10(-5) M) but was blocked by Mn2+ (10(-3) M), D-600 (1 X 10(-7) g/ml) and the specific A II antagonists Sar1-Ala8-angiotensin II (P-113) (5.0 X 10(-5) M) and Sar1-Ile8-angiotensin II (5.28 X 10(-5) M). We conclude that the positive inotropic effect of A II on the myocardium is due to its ability to increase transmembrane ion movements in or through the cell membrane. The ability of Mn2+ and D-600 to block this effect suggests that this ion movement is via the so-called "slow channels."     

Location of nucleolar organizing regions on the chromosomes of the Syrian hamster (Mesocricetus auratus) and the Djungarian hamster (Phodopus sungorus)

Terminal chromosome associations ("satellite associations") in ASG-banded preparations have been used to determine the number and location of staellites in the Syrian hamster (Mesocricetus auratus) and the Djungarian hamster (Phodopus sungorus). Five pairs of satellites are found in the former and four in the latter. Nucleolar organizing regions (NORs) were visualized with the Ag-AS silver precipitation technique, and their number and position corresponded exactly with the number and position of satellites in Phodopus, where positive chromosome identification can be made in the absence of banding. Numerical agreement is exact in Mesocricetus as well, and the morphology of the silver-tagged NOR-bearing chromosomes strongly suggests that corrrespondence also occurs in this species.     

Influence of inorganic soil components and humic substances on the mechanochemical removal of pentachlorophenol

The influence of the presence of humic acids of different origin on the catalytic transformation of pentachlorophenol (PCP) by two metal oxides, ferrihydrite and birnessite, using mechanochemical contact was studied. The mechanochemical dry contact with light grinding of PCP and birnessite was more effective than that observed in the presence of ferrihydrite (approximately 70 and 40% of PCP removal after 20 days, respectively). The removal of PCP by birnessite and ferrihydrite was significantly enhanced in the presence of humic acid (HA). The different origin of humic acids (agricultural, forest and commercial) showed a slight influence on the overall reaction. The commercial HA, when mixed with birnessite, was more effective in reducing PCP concentration compared to the others two HAs investigated. The transformation process in the presence of birnessite with and without HA was accompanied by the release of four chlorides per molecule of PCP transformed.     

URBAN POLICY,PRESS, AND PLACE: CITY‐MAKING IN FLORIDA'S MIAMI‐DADE COUNTY

This article enhances the notion of city‐making by explicating its communicative processes and functions within the press. Through a quantitative content analysis and qualitative textual analysis of Miami Herald news coverage related to incorporation and annexation policies and practices over a period of 3 years, we argue for a stronger implication of the press in coverage of local policy‐ and place‐making. Through a quantitative content analysis of 437 articles from the Miami Herald about communities affected by incorporation and annexation and a qualitative textual analysis of 51 articles related to general coverage of geographic policy‐making in Miami‐Dade County over a 3‐year period, we argue that this coverage reveals the press as being a central feature and function of policy‐making through the lens of city‐making.