Analysis of odontoblast gene expression using a novel approach,laser capture microdissection

Studying the mechanisms of molecular interactions in developing tissues demands sensitive molecular biological in vivo and in vitro techniques. Laser capture microdissection (LCM) allows for the isolation of mRNA in histological sections even from single cells, thus enabling the identification of in vivo gene expression products in closely circumscribed tissue areas. The aims of this study were to assess the optimal fixation, processing, and staining conditions to retrieve RNA from microdissected odontoblasts. Fluorometric assays and RT-PCR analysis of alpha 1(I) collagen, dentin sialophosphoprotein (Dspp), and osteocalcin (OC) confirmed that the total RNA isolated from day 0 and day 3 captured odontoblasts was sufficient in quantity and quality. Our results indicate that individual odontoblasts obtained by LCM are morphologically intact and chemically unaltered, allowing accurate molecular and biochemical analyses.     

Hip fractures among infertile women

A retrospective cohort study was conducted in a population-based inception cohort of 1,157 Olmsted County, Minnesota, women with infertility (failure to conceive after 1 year despite intercourse without contraception) that was first diagnosed at the Mayo Clinic (Rochester, Minnesota) between 1935 and 1964. In this relatively young cohort, 31 hip fractures were observed during 35,849 person-years of follow-up; 36.5 had been expected (standardized incidence ratio = 0.85, 95% confidence interval 0.58-1.20). Standardized incidence ratios did not differ by type or cause of infertility. The data suggested that women with consistently irregular menses may have a greater risk of hip fracture. This finding should be confirmed by additional studies with longer follow-up periods and with assessment of other fracture outcomes.     

The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954)--I. Purification and properties of a nitroreductase enzyme from Escherichia coli--a potential enzyme for antibody-directed enzyme prodrug therapy (ADEPT).

A nitroreductase enzyme has been isolated from Escherichia coli B. This enzyme is an FMN-containing flavoprotein with a molecular mass of 24 kDa and requires either NADH or NADPH as a cofactor. Partial protein sequence analysis showed extensive homology with the "classical nitroreductase" of Salmonella typhimurium and a nitroreductase induced in Enterobacter cloacae. In common with the Salmonella enzyme, the E. coli B enzyme is capable of reducing nitrofurazone. The E. coli nitroreductase is also capable of reducing the anti-tumour agent CB1954 [5-(aziridin-1-yl)-2,4-dinitrobenzamide], a property shared with the mammalian enzyme DT diaphorase [NAD(P)H dehydrogenase (quinone)] as isolated from Walker cells. The reduction of CB1954 by the E. coli enzyme results in the generation of cytotoxic species. Both enzymes also share the properties of being able to reduce quinones and are both inhibited by dicoumarol. The nitroreductase is a more active enzyme against CB1954 (kcat = 360 min-1) than Walker DT diaphorase (kcat = 4 min-1) and also has a lower Km for NADH (6 vs 75 microM).     

Community Health In a Mountain Neighborhood

To test the hypothesis that during high periods of carbon monoxide (CO) pollution, persons with acute cardiovascular disease would be adversely affected, we have studied case fatality rates for patients admitted with myocardial infarction (Ml) to 35 Los Angeles hospitals during 1958. The results indicate that there is an increased Ml case fatality rate in “high”-pollution areas and that this difference is only evident during periods of relatively increased CO pollution.

At the present time, our interpretation of these findings is that an association could exist between ME case fatality rate and atmospheric CO pollution but we cannot draw any firm conclusions about causality at this time.