Hippocrates on necrotising fasciitis.
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Biomedical subjects
Publications and source records attributed to J Aitken.
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Although the generation of reactive oxygen species is an activity normally associated with phagocytic leucocytes, mammalian spermatozoa were, in fact, the first cell type in which this activity was described. In recent years it has become apparent that spermatozoa are not the only nonphagocytic cells to exhibit a capacity for reactive oxygen species production, because this activity has been detected in a wide variety of different cells including fibroblasts, mesangial cells, oocytes, Leydig cells, endothelial cells, thyroid cells, adipocytes, tumour cells and platelets. Since the capacity to generate reactive oxygen species is apparently so widespread, the risk-benefit equation for these potentially pernicious molecules becomes a matter of intense interest. In the case of human spermatozoa, the risk of manufacturing reactive oxygen metabolites is considerable because these cells are particularly vulnerable to lipid peroxidation. Indeed, there is now good evidence to indicate that oxygen radicals are involved in the initiation of peroxidative damage to the sperm plasma membrane, seen in many cases of male infertility. This risk is off-set by recent data suggesting that superoxide anions and hydrogen peroxide also participate in the induction of key biological events such as hyperactivated motility and the acrosome reaction. Thus, human spermatozoa appear to use reactive oxygen species for a physiological purpose and have the difficult task of ensuring the balanced generation of these potentially harmful, but biologically important, modulators of cellular function.
In this study, we have examined the relationship between creatine phosphokinase (CPK), a biochemical measure of human sperm quality (Huszar et al., 1988a,b, 1990; Huszar and Vigue, 1994), and a marker for the presence of residual cytoplasm in human spermatozoa, glucose-6-phosphate dehydrogenase (G6PDH). We then determined whether the diagnostic potential of these enzymes was related to the capacity of the sperm suspensions to generate reactive oxygen species (ROS) and/or the presence of leukocytes and precursor germ cells. Across the data set as a whole, G6PDH and CPK were highly correlated with each other and, to a lesser extent, with the generation of ROS. Contamination of the sperm suspensions with leukocytes might have contributed to these associations, since the presence of such cells was also significantly correlated with CPK, G6PDH, and ROS. However, even after the leukocytes had been carefully removed, G6PDH was still highly correlated with CPK (r = 0.794), indicating that both criteria were providing similar information of the cytosolic component of human sperm suspensions. In the absence of leukocyte contamination, CPK and G6PDH activities were also correlated with the presence of precursor germ cells, and this association may, in part, explain the diagnostic value of these criteria. An additional component of their prognostic value may be reflected in the statistically significant association observed between G6PDH activity and ROS generation. A possible mechanism for such an association is suggested, which should be amenable to experimental verification.
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We conducted a hospital-based case-control study in Brisbane, Queensland, to investigate the specific determinants of basal cell carcinoma (BCC), as distinct from squamous cell carcinoma (SCC), with emphasis on ancestry, residential history, pigmentary characteristics, sun sensitivity and other constitutional factors. The sample was recruited from a dermatology outpatient clinic during an eight-week period in 1991, and comprised 51 incident or recently diagnosed cases of BCC, and 112 randomly selected controls with no known history of BCC or SCC. Twenty-six cases with both BCC and SCC were analysed separately. We found no risk factor specific to BCC which might explain its extremely high prevalence. The strongest risk factors for BCC were advanced age, male sex and a propensity to freckle (independent of skin colour and tendency to burn), all of which have previously been observed for SCC. A finding not previously reported was an apparent protective effect of increasing body mass, specific to BCC alone.
Sputum samples submitted to the microbiology laboratory from general medical and respiratory wards were monitored for Moraxella catarrhalis on a prospective basis. All isolates were typed by restriction endonuclease typing. Nosocomial spread was found both by the clustering of cases and typing of isolates. Sampling of the environment of some cases was performed. Seven out of 37 samples revealed environmental contamination. Sampling for persistence of the organism in the environment was positive on one occasion out of 13. Evaluation of acquisition of M. catarrhalis in relation to length of stay showed that the average length of stay of a case with M. catarrhalis was considerably longer than average patient stay without M. catarrhalis. Four patients had two isolates available for typing. The type of M. catarrhalis was different on the second occasion to that on the first. Nosocomial spread of M. catarrhalis in the setting of general medical and respiratory wards was found to occur in the winter months.
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C19 steroids are converted to estrogens in a number of tissues by a specific form of cytochrome P450, namely aromatase cytochrome P450 (P450arom). Adipose tissue is the principal site of estrogen formation in postmenopausal women. Aromatase activity as well as P450arom transcripts primarily reside in the stromal cell component of the adipose tissue. Studies designed to investigate whether increased local aromatase activity in breast adipose tissue influences the growth of breast cancers have yielded discrepant results. In an attempt to clarify this controversy, adipose tissue was obtained from the four breast quadrants at the time of mastectomy (n = 13) performed for removal of a tumor. Breast fat P450arom messenger RNA levels were quantified and compared between the four quadrants within each specimen using competitive polymerase chain reaction after reverse transcription in which 10 micrograms human adipose total RNA together with 1 pg rat complementary RNA (internal standard) were reverse transcribed and coamplified. In 9 out of 13 patients (69%), highest P450arom transcript levels colocalized to the quadrants bearing tumors. This correlation was statistically significant (P < 0.001). The regional distribution of P450arom transcripts in breast adipose tissue of disease-free individuals, obtained during reduction mammoplasty (control group, n = 9), did not favor any particular region of the breast. We also quantified by morphometry the histological components of the adipose tissue samples from each quadrant in mastectomy specimens. The distribution of stromal cells significantly correlated with the distribution of P450arom transcript levels, in that quadrants containing highest proportions of stromal cells matched to highest transcript levels (P < 0.01). Although the quadrants bearing tumors contained the highest percentage of stromal cells, this correlation was not statistically significant. The adipose tissue surrounding a breast tumor displays increased estrogen biosynthesis, which may promote tumor growth. It is further suggested that the distribution of stromal cell components in breast adipose tissue gives rise to locally elevated P450arom expression, which in turn may favor neoplastic development and growth in these predisposed areas of the breast. The correlation between the presence of a tumor and elevated P450arom levels in the proximal adipose tissue is independent of tumor size, node involvement, histological type or grade, estrogen/progesterone receptor status, DNA index, or S-phase fraction.
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The aromatase enzyme complex is responsible for the conversion of C19 steroids to estrogens. Aromatase activities ranging from moderate to very low have been measured in human fetal tissues. The inability to demonstrate aromatase cytochrome P450 (P450AROM) messenger RNA (mRNA) in several fetal tissues by northern blotting has been attributed to low levels of specific message. In order to identify and compare P450AROM mRNA levels in fetal tissues, we developed a specific competitive polymerase chain reaction amplification technique. This reaction uses coamplification of a rat P450AROM complementary RNA to normalize differences in amplification efficiencies. Using this technique, P450AROM mRNA was identified in all fetal tissues studied including; liver, lung, brain, skin, intestine, kidney, spleen, and heart. Fetal liver contained far more P450AROM mRNA per total RNA than any other tissues studied. Fetal brain and intestine also tended to have slightly higher levels than other tissues.
Local production of estrogen in breast tissue may influence the growth of breast cancers. Peripheral conversion of C19 steroids to estrogens is catalyzed by the aromatase enzyme complex which is comprised of a specific form of cytochrome P450, aromatase cytochrome P450 (P450AROM) and the flavoprotein, NADPH-cytochrome P450 reductase. To evaluate P450AROM mRNA levels in breast tissue, a specific competitive polymerase chain reaction amplification procedure was devised. In this method, a rat P450AROM complementary RNA is coamplified as an internal standard in order to compare amplification reactions. The amplification products are recognized by hybridization with 32P-labeled oligonucleotides specific for each species. Densitometry is used to quantitate autoradiographs. Initial studies using RNA from whole breast tissue obtained from reduction mammoplasty revealed linearity of the relationship between the densitometer signal from the human amplification product and total RNA concentration. Breast tissue was then separated into a floating adipocyte fraction and a pelleted fraction containing the other cellular elements by collagenase digestion and centrifugation. Comparison of specific content of aromatase amplification product per unit weight of RNA extracted from adipocytes and pelleted cells revealed considerably higher levels in the RNA from the nonadipocyte fraction. Immunocytochemical characterization of this fraction revealed the presence of several cell types including macrophages, ductal epithelial cells, and endothelial cells, but primary cells of stromal origin.
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Estrogen production by adipose tissue has been implicated in the etiology of such human cancers as endometrial and breast cancer. Estrogen production by adipose cells is subject to complex multifactorial regulation by a number of growth factors and cytokines, including those produced by breast cancer cells. In order to understand the mechanisms responsible for aromatase regulation, the structural gene encoding aromatase cytochrome P-450 (P-450AROM) was isolated from human genomic DNA. The gene spans at least 70 kb and is comprised of 10 exons, the first of which is untranslated. DNA sequence analysis indicates that the gene has a putative TATA (ATAAAA) sequence at -23 bp and putative CAAT binding sequences beginning at -41, -67, and -83 bp, that constitute a promoter region responsible for expression in placenta. However, this promoter does not appear to be responsible for expression in adipose, which may therefore be under the control of another, tissue-specific, promoter. Use of Polymerase Chain Reaction (PCR) technology has allowed for determination of expression of P-450AROM in samples of breast adipose. Preliminary results indicate that expression is highest in the upper lateral region, similar to the site of most frequent localization of tumors.
A synchronous occurrence of large bowel adenocarcinoma and extragenital malignant mixed mesodermal tumour (MMMT) is reported. This case represents the sixth extragenital MMMT reported in the literature.
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Following a 20 patient pilot investigation for Campylobacter pyloridis in selected gastric biopsy specimens, 56 consecutive patients subjected to routine gastroscopies were further examined. 14/56 had positive gram stains for spiral gram negative bacilli in gastric biopsy imprints and 4/14 had positive culture for this organism. All 14 positive specimens showed histological gastritis of varying degrees (grade 3 inflammation). The methodology, significance and implications of these findings are discussed in this first New Zealand report of C pyloridis in the stomach.
A fixed schedule for ovarian stimulation and follicular aspiration, previously used in our department for research purposes, was modified in an attempt to increase the recovery and cleavage rates of the oocytes. Three different clomiphene regimens were used to stimulate the ovaries of normal volunteer women requesting laparoscopic sterilization (50 mg and 150 mg daily for 5 days, and 50 mg daily for 10 days). Oocytes were recovered from 83% of the aspirated follicles, i.e. 1.6 oocytes/patient: 65% of the oocytes cleaved after in-vitro fertilization and, on average, 1.0 cleaving egg was obtained per patient. There were no differences in the recovery and cleavage rates between the three clomiphene regimens. It is concluded that a sufficient number of cleaving embryos for research purposes can be generated with the present fixed regimen, which offers little inconvenience to the volunteers as the day of egg recovery can be predicted some time in advance.