17 beta-estradiol hydroxylation catalyzed by human cytochrome P450 1B1

The 4-hydroxy metabolite of 17 beta-estradiol (E2) has been implicated in the carcinogenicity of this hormone. Previous studies showed that aryl hydrocarbon-receptor agonists induced a cytochrome P450 that catalyzed the 4-hydroxylation of E2. This activity was associated with human P450 1B1. To determine the relationship of the human P450 1B1 gene product and E2 4-hydroxylation, the protein was expressed in Saccharomyces cerevisiae. Microsomes from the transformed yeast catalyzed the 4- and 2-hydroxylation of E2 with Km values of 0.71 and 0.78 microM and turnover numbers of 1.39 and 0.27 nmol product min-1.nmol P450-1, respectively. Treatment of MCF-7 human breast cancer cells with the aryl hydrocarbon-receptor ligand indolo[3,2-b]carbazole resulted in a concentration-dependent increase in P450 1B1 and P450 1A1 mRNA levels, and caused increased rates of 2-, 4-, 6 alpha-, and 15 alpha-hydroxylation of E2. At an E2 concentration of 10 nM, the increased rates of 2- and 4-hydroxylation were approximately equal, emphasizing the significance of the low Km P450 1B1-component of E2 metabolism. These studies demonstrate that human P450 1B1 is a catalytically efficient E2 4-hydroxylase that is likely to participate in endocrine regulation and the toxicity of estrogens.     

The influence of growth hormone (rhGH) therapy on tooth formation in idiopathic short statured children

The purpose of this preliminary study was to evaluate tooth formation in children with idiopathic short stature, before and during treatment with recombinant growth hormone (rhGH). Twenty-nine short-statured children ages 6 to 13 years were assigned into two treatment groups, an "experimental" group (n = 18), which received rhGH, and a "control" group (n = 11), which was observed for 1 year before commencing rhGH treatment. Clinical and radiographic records were obtained at the initial, year 1, and year 2 visits. Tooth formation and stature were assessed by calculating Z-scores, appropriate for the age and gender of each child. Delta-Z scores, which measure the change in Z-score over time, were also calculated between annual visits. Height was measured and recorded every 3 months, and Z-score statural norms for age and gender were derived from the 1977 National Center for Health Services national probability sampling. Tooth formation standards were derived from Moorrees et al. A matched control sample for tooth development was derived from untreated children. Tooth formation was initially delayed although the degree of reduction in stature exceeded the initial degree of delay in tooth formation. During this 2-year study, rhGH therapy had a significant influence on acceleration or gain in stature, but did not have a significant influence on tooth formation.     

An image processing scheme to quantitatively extract and validate hyoid bone motion based on real-time ultrasound recordings of swallowing

An image processing technique with associated hardware was developed to quantitatively extract hyoid bone motion from realtime submandibular ultrasound images recorded during the swallowing act. Videofluorographic imaging, the "gold standard" of swallowing studies, was recorded simultaneously and synchronized to the ultrasound. Hyoid position obtained from the ultrasound was validated based on the videofluorography using personal computer-based image processing methods.     

Influence of route of administration on [3H]-camptothecin distribution and tumor uptake in CASE-bearing nude mice: whole-body autoradiographic studies

The small subunit of the bovine mitochondrial ribosome forms a tight complex with mRNAs. This [28 S:mRNA] complex forms as readily on circular mRNAs as on linear mRNAs indicating that a free 5' end on the mRNA is not required for the interaction observed. The effects of monovalent cations on the equilibrium association constant and on the forward and reverse rate constants governing this interaction have been determined. Monovalent cations have a strong effect on the forward rate constant. Increasing the KCl concentration from 1 mM to 100 mM reduces kon by nearly 100-fold. Monovalent cations have only a small effect on the reverse rate constant, koff'. Analysis of these data indicates that the rate laws governing the formation and dissociation of the [28 S:mRNA] complex cannot be deduced from the chemical equation. This observation suggests that there are "hidden intermediates' in the formation and dissociation of this complex. The implications of these observations are discussed in terms of a model for the interaction between the mitochondrial 28 S subunit and mRNAs.     

Cardiac surgical implications of calcium dyshomeostasis in the heart

The prevalence of coronary artery disease renders myocardial ischemia a leading cause of morbidity and mortality. Both cardiac bypass operations and cardiac transplantation cause myocardial ischemia and reperfusion injury. Intracellular calcium transport and regulation are of paramount importance in both normal and pathologic myocardial states. Calcium regulation is integral to nearly every myocyte function, from early development to senescence. Normal intracellular calcium-mediated excitation-contraction coupling and abnormal patterns of calcium regulation leading to systolic/diastolic dysfunction are now therapeutically accessible to the cardiac surgeon. Additionally, altered Ca2+ transport protein gene expression is a mechanism of myocardial dysfunction. Therapeutic strategies involve receptor-mediated transduction of signals to intracellular metabolic sites. Evidence implicates protein kinase C as well as a potential therapeutic role for Ca2+. The potential for pharmacologic access to this protective state has abundant clinical appeal. The protective state (cardiac "preconditioning") is transient but is amenable as therapy against operation-related ischemic events.     

Influence of viraemia and genotype upon serological reactivity in screening assays for antibody to hepatitis C virus

Ganglioside GM2, expressed on the surface of some human cancers, is a promising target for immune therapy, since GM2 antibodies are cytotoxic, can be induced in humans by vaccination, and the presence of GM2 antibodies is associated with a better prognosis in melanoma patients. In our efforts to induce long-lived, cytotoxic GM2 antibodies, we investigated lipopolysaccharides (LPS) containing "GM2-like" oligosaccharides. LPS were prepared from Campylobacter jejuni serotypes O:1, O:23, or O:36 (all sharing the oligosaccharide structure GalNAcbeta1-4Gal(113NeuAc)-Hex with ganglioside GM2), and tested for their ability to induce GM2-reactive antibodies. Immunization of NZW rabbits (2 animals per vaccine) with LPS from C. jejuni serotype O:1 in Freund's adjuvant resulted in production of high-titer IgG antibodies reactive with purified bovine brain GM2 in ELISA, dot-blot immune strains and immune thin-layer chromatography, and with GM2 derived from various human tumors by immune thin-layer chromatography. These rabbit antibodies bound to cancer cell lines expressing GM2 on their cell surface, as determined by mixed hemadsorption assays, mediating strong antibody-dependent cellular cytotoxicity (ADCC) with tumor cells expressing cell-surface GM2. Antibodies induced by vaccination with C. jejuni serotype O:1 were higher-titer (IgG ELISA titer > 1:60,000) than antibodies induced by immunization with purified GM2 (IgG ELISA titer > 1:200). Immunization with LPS from C. jejuni serotype O:36 resulted in production of moderately high-titer IgM and low-titer IgG GM2 antibodies. Immunization with LPS from C. jejuni serotype O:23 did not elicit GM2-reactive antibodies. No clinical symptoms were observed in animals immunized with these LPS preparations, with purified GM2 ganglioside, or with LPS derived from C. jejuni serotype O:19 (containing a GM1-like oligosaccharide). Our results indicate that lipopolysaccharides sharing carbohydrate epitopes with gangliosides may be useful immunogens for inducing antibodies to ganglioside antigens.     

Impacts of ice cover on local scour around semi-circular bridge abutment简

The presence of ice cover in winter can significantly change the flow field around bridge abutments, which can also cause a different local scour pattern. To investigate the impacts of ice cover, results from a recent flume experiments were presented. Smooth and rough ice covers were created to investigate the impacts of ice cover roughness on the scour geometry around the semi-circular abutment. Three bed materials were used, with 50D s of 0.58 mm, 0.50 mm, 0.47 mm respectively. Scour volume and scour area were calculated. It was found that the maximum scour depth was located 75o inclined to the flume wall. Under rough ice cover, the scour area and scour depth were the largest. An empirical equation on the maximum scour depth was also developed.     

Vascular endothelial growth factor: crystal structure and functional mapping of the kinase domain receptor binding site

Vascular endothelial growth factor (VEGF) is a homodimeric member of the cystine knot family of growth factors, with limited sequence homology to platelet-derived growth factor (PDGF) and transforming growth factor beta2 (TGF-beta). We have determined its crystal structure at a resolution of 2.5 A, and identified its kinase domain receptor (KDR) binding site using mutational analysis. Overall, the VEGF monomer resembles that of PDGF, but its N-terminal segment is helical rather than extended. The dimerization mode of VEGF is similar to that of PDGF and very different from that of TGF-beta. Mutational analysis of VEGF reveals that symmetrical binding sites for KDR are located at each pole of the VEGF homodimer. Each site contains two functional "hot spots" composed of binding determinants presented across the subunit interface. The two most important determinants are located within the largest hot spot on a short, three-stranded sheet that is conserved in PDGF and TGF-beta. Functional analysis of the binding epitopes for two receptor-blocking antibodies reveal different binding determinants near each of the KDR binding hot spots.     

Aetiology and occurrence of diving injuries. A review of diving safety

This paper examines multifaceted aspects of diving entries into water which are the cause of many critical injuries (costed at $A150 million) and therefore have important safety ramifications. Wedge and compression fractures are most commonly found in the cervical area of the spine with off-centre impacts with the pool or sea bottom. Diving-related injuries range from 2.3 in a South African study to 21% of spinal cord injuries in Poland. Alcohol and diving do not mix because of diminished awareness and information processing. Children aged under 13 years suffer fewer cervical injuries (1 to 4%), but complication rates are relatively high for this group. Sports trauma (diving-related in particular) is one of the more prevalent causes of spinal cord injury in children aged 6 to 15 years. The highest incidence occurs among those aged 10 to 14, followed by the group aged 5 to 9 years. This contradicts the common perception that 15-to 19-year-olds comprise the highest risk group. Boys are more frequently injured, and swimming pools are more common as an injury location then is the case with adults. The role played by water depth has been conclusively ascertained; technique, and therefore education, appear to be more important considerations in injury prevention. Although 89% of injuries occur in water < 1.52m, injuries are rare in water of 0.46 to 0.61m. Care with pool design to avoid sudden depth changes and the resultant "spinal wall' is necessary. Minimum depth values for diving vary from 1 to 1.52 m. Velocities and angles of entry are considered to ascertain the body's decelerative capacity upon entry. The scoop, racing start dive has been shown to require at least 1.22 m of water even when practised by trained divers; the risks involved must therefore be weighed against the fact that it may be no faster than more conventional dives. While it may be safe to perform kneeling and crouching dives into shallowers water, standing dives by untrained divers require a greater margin of error. Lack of education is an issue which needs to be addressed and this paper makes recommendations for safety practices such as steering up to the surface, head protection with the arms and only diving when absolutely necessary.