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Biomedical subjects

A S Field

Publications and source records attributed to A S Field.

At least 19 recordsLinked to original sources

Hormone replacement therapy use dramatically increases breast oestrogen receptor expression in obese postmenopausal women.

BACKGROUND: It is known that use of hormone replacement therapy (HRT) by postmenopausal women increases the risk of breast cancer. METHOD: In this study, oestrogen receptor (ER)-alpha expression is examined using standard immunoperoxidase technique. RESULTS: Normal breast samples of 11 Australian postmenopausal women have been included in the ER-alpha study; the result showed a strong correlation (r(2) = 0.80) between ER-alpha expression in normal breast epithelial cells and body mass index (BMI) in normal women who currently use HRT. CONCLUSION: This finding confirms that the possibility of increased risk of breast cancer associated with increased ER-alpha expression in normal breast epithelial cells, in turn associated with high BMI and the use of HRT.

Adult↗

Suggestions for HER-2/neu testing in breast carcinoma, based on a comparison of immunohistochemistry and fluorescence in situ hybridisation.

The arrival of Herceptin (Trastuzumab), an antibody against the HER-2 oncogene found in a proportion of breast carcinomas and other carcinomas, has emphasised the need for a standardised technique for demonstrating overexpression of HER-2. We compared the Dako A485 antibody and Dako HercepTest kit (HT) on a series of 122 breast carcinomas. Fluorescence in situ hybridisation (FISH) (Vysis) was performed on all cases with positive or equivocal immunohistochemical results. The Dako A485 showed HER-2 overexpression in 53% of carcinomas, while the HT showed 21% positive (HT 2+ 8%, HT 3+ 13%) and 79% negative (HT 0 67%, HT 1+ 12%). FISH for HER-2 gene amplification on all the HT 1+ and HT 2+ cases was negative, whereas FISH analysis of all HT 3+ cases was positive, with the exception of one case which could not be analysed for technical reasons. When histological subtype was analysed, only grade 3 infiltrating duct carcinomas were FISH-positive, suggesting that histological grading and subtyping may be able to triage carcinomas suitable for HER-2 testing. We suggest that the HT or a similar standardised immunohistochemical study for HER-2 can be used to screen breast carcinomas. We then recommend FISH where the carcinoma is HT 2+. FISH may also be appropriate in high grade, HT 1+ carcinomas where there are doubts regarding optimal tissue fixation or block storage conditions.

Antibodies, Monoclonal↗

False cerebral activation on BOLD functional MR images: study of low-amplitude motion weakly correlated to stimulus.

BACKGROUND AND PURPOSE: Movements of the participant during blood oxygen level-dependent (BOLD) functional MR imaging cerebral activation studies are known to produce occasionally regions of false activation, especially when these movements are relatively large (>3 mm) and highly correlated with the stimulus. We investigated whether minimal (<1 mm), weakly correlated movements in a controlled functional MR imaging model could produce false activation artifacts that could potentially mimic regions of true activation in size, location, and statistical significance. METHODS: A life-size brain phantom was constructed by embedding vials of a dilute carboxylic acid solution within a gadolinium-doped gelatin mold. Imaging was performed at 1.5 T using a 2D spiral sequence (3,000/5 [TR/TE]; flip angle, 88 degrees; matrix, 64 x 64; field of view, 24 cm; section thickness, 5 mm). Controlled, in-plane, submillimeter movements of the phantom were generated using a pneumatic system and were made to correlate with a hypothetical "boxcar" stimulus over the range 0.31 < r < 0.96. Regions of false activation were sought using standard statistical methods (SPM96) that excluded phantom edges and accounted for spatial extent (regions tested at P < .05, corrected for multiple comparisons). A similar experiment was performed on a resting volunteer. RESULTS: The pneumatic system provided motion control with average in-plane displacements and rotations of 0.74 mm and 0.47 degrees, respectively, in the 18 data sets analyzed. No areas of false activation in the phantom were identified for poorly correlated motions (r < 0.52). Above this level, false activations occurred with increasing frequency, scaling in size and number with the degree of motion correlation. For motions with r > 0.67, areas of false activation were seen in every experiment. For a statistical threshold of P = .001, the median number of falsely activated regions was 3.5, with a mean size of 71.7 voxels (approximately 5 cc). Areas of possibly false activation of average size 72.5 voxels resulting from passive motion of the resting human participant were observed in two of four experiments. CONCLUSION: Participant movements of 1 mm or less that are only modestly correlated with a blocked stimulus paradigm can produce appreciable false activation artifacts on BOLD functional MR imaging studies, even when strict image realignment methods are used to prevent them.

Artifacts↗

Cyclin D1 protein is overexpressed in hyperplasia and intraductal carcinoma of the breast.

The cell cycle regulatory gene cyclin D1 is a candidate oncogene in breast cancer. It is overexpressed in 30-50% of invasive primary breast cancers and plays a key role in mediating mitogenic responses to steroids and growth factors in breast cancer cells in vitro. Because the role of cyclin D1 in the proliferative and early noninvasive stages of breast cancer is largely unknown, we examined normal breast epithelium (NBE), proliferative disease (PD), ductal carcinoma in situ (DCIS), and invasive carcinoma (IC) to evaluate the timing and possible importance of cyclin D1 expression in the development of breast cancer. Using immunohistochemistry, we examined cyclin D1 protein expression in 471 breast tissue samples. A quantitative scoring system for immunohistochemistry based on percentage of positive cells was developed that correlated with Western blot analysis of antigen concentration in paired samples (r2 = 0.91, P = 0.003). A sample was considered positive if >5% of relevant epithelial cells demonstrated nuclear staining. Cyclin D1 positivity was observed in 11.7% (7 of 60) samples of NBE, 25% (11 of 44) of PD without atypia, 39.4% (13 of 33) of atypical ductal hyperplasia, 43.6% (17 of 39) of low-grade DCIS, 47.9% (23 of 48) of high-grade DCIS, and 48.3% (99 of 205) of IC. Cyclin D1 expression was significantly higher in PD than NBE (P = 0.006) and in DCIS than PD (P = 0.038). There was no significant increase from DCIS to IC (P = 0.52). The increase in cyclin D1 expression in the overall progression from NBE to IC was also highly significant (P = 0.0001). Therefore, cyclin D1 expression was detected at levels significantly greater than in NBE in the earliest proliferative epithelial lesions of the breast with a further significant increase accompanying the progression to any form of cancer. This suggests that overexpression of cyclin D1 protein is important at the earliest stages of breast oncogenesis and continues to have a crucial role throughout the development of malignancy.

Breast Neoplasms↗

Development and ultrastructure of Trachipleistophora hominis n.g., n.sp. after in vitro isolation from an AIDS patient and inoculation into athymic mice.

Continuous culture was achieved in several cell lines of a microsporidium obtained from the skeletal muscle of an AIDS patient. Development in COS-1 and RK13 cells was prolific. Spores from the original biopsy were also inoculated into athymic mice by i.m. and i.p. routes. Infection was found in several organs as well as in skeletal muscle after a few weeks. All stages were surrounded by an electron-dense surface coat. Meronts had 2-4 nuclei and divided by binary fission. In sporogony the surface coat became separated from the plasma membrane to form a sporophorous vesicle, within which division into sporoblasts was effected by repeated binary fissions. The number of sporoblasts (and later spores) within the sporophorous vesicles varied from 2 to > 32 and the sizes of the vesicles varied, according to the number of spores contained therein, from 5 microns diameter to 14.0 x 11.0 microns. Spores measured 4.0 x 2.4 microns and had a prominent posterior vacuole. The parasite differs from the genus Pleistophora in that it does not form multinucleate sporogonial plasmodia and that the sporophorous vesicle enlarges during sporogony and its wall is not a multilayered structure. It is proposed to place it in a new genus and species Trachipleistophora hominis n.g., n.sp.

AIDS-Related Opportunistic Infections↗

Myositis associated with a newly described microsporidian, Trachipleistophora hominis, in a patient with AIDS.

Microsporidia are zoonotic protozoa which were rare human pathogens prior to 1985, when Enterocytozoon bieneusi was described in human immunodeficiency virus-infected patients with chronic diarrhea. Another species, Encephalitozoon (Septata) intestinalis, is associated with diarrhea and chronic sinusitis, and approximately 25 cases have been reported in the literature. However, other microsporidial infections in human immunodeficiency virus-infected patients remain extremely rare. We report the first case of a Pleistophora sp.-like microsporidian infection presenting as a progressive severe myosotis associated with fever and weight loss. The organism was demonstrated by light microscopy and electron microscopy in corneal scrapings, skeletal muscle, and nasal discharge. Electron microscopy showed an electron-dense surface coat with "sunflare"-like projections surrounding all stages of development of meronts (two to four nuclei, dividing by binary fission), sporonts, and sporoblasts. Division of sporonts, in which sporonts separate from the thick outer coat, creating a sporophorous vesicle, is by binary fission, differentiating this organism from Pleistophora sp. The spore measures 4.0 by 2.5 microns and has a rugose exospore. A new genus and species, Trachipleistophora hominis, has been established for this parasite. The patient was treated with albendazole, sulfadiazine, and pyrimethamine, and the clinical symptoms resolved.

AIDS-Related Opportunistic Infections↗

Disseminated microsporidiosis due to Septata intestinalis in nine patients infected with the human immunodeficiency virus: response to therapy with albendazole.

Disseminated microsporidiosis due to the newly described species Septata intestinalis in nine patients infected with human immunodeficiency virus is described. All patients were male homosexuals; the mean age was 41 years (range, 35-58 years). They were all severely immunocompromised, with a mean CD4 lymphocyte count of 15/mm3 (range, 0-32/mm3). Infection by S. intestinalis was seen in duodenal biopsy specimens from all patients, and dissemination was demonstrated by the presence of microsporidial spores in urine (9 of 9 patients), sinonasal secretions and/or nasal mucosal biopsy specimens (6 of 6), and sputum (6 of 6). Seven patients were treated with albendazole (400 mg twice daily), resulting in significant dissipation or complete resolution of diarrhea for six patients and abatement of symptoms for the six patients with chronic rhinosinusitis. There was a parallel parasitological response, with clearance of S. intestinalis infection from almost all sites.

AIDS-Related Opportunistic Infections↗

Microsporidia in the small intestine of HIV-infected patients. A new diagnostic technique and a new species.

OBJECTIVES: To determine whether microsporidian infections occur in Australian patients infected with human immunodeficiency virus (HIV), to assess the incidence, and to discuss microscopic detection methods. DESIGN AND PATIENTS: 180 consecutive HIV-infected patients (109 with chronic diarrhoea and 71 with other indications) underwent upper gastrointestinal tract endoscopy and pinch biopsies of the second part of the duodenum. The biopsies were handled by a protocol: four levels, with haematoxylin and eosin stain (H&E) at each level, periodic acid Schiff reagent after diastase (DiPAS) and auramine stain at the second level, and Warthin-Starry (WS) stain and cytomegalovirus early antigen immunoperoxidase study at the third level. Electron microscopy was carried out on samples from the first 95 patients, and thereafter from selected patients. SETTING: The patients came from the HIV Medicine Unit of a teaching hospital and from the practice of a gastroenterologist. MAIN OUTCOME MEASURES: Diagnosis of microsporidia was to based on the H&E stain, with electron microscopy as the definitive test because the microsporidia are often difficult to see with H&E. Empirically, the WS stain was found to stain the microorganisms and it replaced electron microscopy during the study as the screening diagnostic test. RESULTS: Microsporidia were present in 36 of the 109 patients with diarrhoea (33%) and one of 71 patients without diarrhoea. The WS stain in all cases showed developing spores in the enterocytes and in four cases in macrophages as well. The H&E stain showed non-specific duodenitis and was not diagnostic in some cases. Electron microscopy on samples from the first 95 consecutive patients showed 100% concordance with the WS stain. In 33 cases, electron microscopy showed the multinucleated plasmodia and the spores of Enterocytozoon bieneusi and in the four cases confirmed the spores in macrophages and showed a new Encephalitozoon-like species with a septate parasitophorous vacuole. Other causes of duodenal infection were cytomegalovirus (11 cases), mycobacteria (8), cryptosporidia (8) and Giardia lamblia (5). CONCLUSION: E. bieneusi was the commonest microorganism found in our series of 180 consecutive patients. The actual prevalence of the two microsporidia species within the HIV-positive population and general community awaits further study. The WS stain provides a sensitive diagnostic test for the presence of E. bieneusi and the new Encephalitozoon-like species, avoiding the cost and potential sampling error of electron microscopy. The detailed ultrastructure and taxonomy of the new species requires further study.

AIDS-Related Opportunistic Infections↗

The effect of pulsed microwaves on passive electrical properties and interspike intervals of snail neurons.

The effects of pulsed microwaves (2.45 GHz, 10 microseconds, 100 pps, SAR: 81.5 kW/kg peak, 81.5 W/kg average) on membrane input resistance and action potential (AP) interval statistics were studied in spontaneously active ganglion neurons of land snails (Helix aspersa), at strictly constant temperature (20.8 +/- .07 degrees C worst case). Statistical comparison with sham-irradiated neurons revealed a significant increase in the mean input resistance of neurons exposed to pulsed microwaves (P < or = .05). Pulsed microwaves had no visible effect on mean AP firing rate; this observation was confirmed by analysis of interspike intervals (ISIs). Using an integrator model for spontaneously active neurons, we found the net input current to be more variable in neurons exposed to pulsed microwaves. The mean input current was not affected. The standard deviation of ISIs and the autocorrelation of the input current were marginally affected, but these changes were not consistent across neurons. Although the observed effects were less obvious than those reported in other studies, they represent evidence of a direct interaction between neurons and pulsed microwaves, in the absence of macroscopic temperature changes. The data do not suggest a single, specific mechanism for such interaction.

Animals↗

Microsporidial keratoconjunctivitis in AIDS.

Microsporidia are rare human opportunistic pathogens that are now increasingly recognised in patients with AIDS, most commonly as an ocular or enteric pathogen. This paper reports a case of microsporidial keratoconjunctivitis in a patient with AIDS which responded to treatment with dibromopropamidine isethionate ointment. The clinical and laboratory features of microsporidial ocular infection are reviewed.

AIDS-Related Opportunistic Infections↗

Carcinoma of the thyroid gland and Gardner's syndrome.

A patient known to be suffering from Gardner's syndrome who developed carcinoma of the thyroid gland is presented. A review of the literature reveals that this relationship is not fortuitous and that it has the following characteristics: female predominance (89%), youth (< 30 years in 78%), papillary form (88%), multicentricity (70%) and thyroid carcinoma preceding diagnosis of hereditary polyposis in 30%. Hereditary polyposis patients affected by familial polyposis coli are at significant risk of developing extracolonic malignant tumours, and require life-long surveillance. Palpation of the thyroid gland and possibly ultrasound examination of the neck should be part of the routine follow up of these patients.

Adult↗

The Warthin-Starry stain in the diagnosis of small intestinal microsporidiosis in HIV-infected patients.

A protocol for the handling of small intestinal biopsies from HIV-infected patients is presented. This protocol includes the Warthin-Starry stain for the detection of microsporidia. This stain has proved a reliable and sensitive diagnostic technique for microsporidial infections as it stains both Enterocytozoon bieneusi and Septata intestinalis in duodenal enterocytes. Because the stain demonstrates Septata intestinalis in lamina propria macrophages as well as enterocytes, it allows for the practical differentiation of these two microsporidial infections. The Warthin-Starry stain has also demonstrated Septata intestinalis in nasal and colonic biopsies in some of these patients. Since the completion of an earlier study, a further 40 cases of Enterocytozoon bieneusi and three cases of Septata intestinalis have been diagnosed in just over 240 consecutive duodenal biopsies from HIV positive patients presenting with diarrhoea and other gastrointestinal complaints. Other opportunistic infections include cytomegalovirus in four cases, mycobacteria in eight cases, cryptosporidia in nine cases, giardia in four cases and Isospora belli in one case. Since the ratio of these opportunistic infections has remained much the same as in the previous study of 180 consecutive duodenal biopsies, we suggest that these rates may reflect the actual prevalence of microsporidial infections in AIDS patients in Sydney, Australia.

AIDS-Related Opportunistic Infections↗

Topographic component (parallel factor) analysis of multichannel evoked potentials: practical issues in trilinear spatiotemporal decomposition.

We describe a substantive application of the trilinear topographic components/parallel factors model (TC/PARAFAC, due to Möcks/Harshman) to the decomposition of multichannel evoked potentials (MEP's). We provide practical guidelines and procedures for applying PARAFAC methodology to MEP decomposition. Specifically, we apply techniques of data preprocessing, orthogonality constraints, and validation of solutions in a complete TC analysis, for the first time using actual MEP data. The TC model is shown to be superior to the traditional bilinear principal components model in terms of data reduction, confirming the advantage of the TC model's added assumptions. The model is then shown to provide a unique spatiotemporal decomposition that is reproducible in different subject groups. The components are shown to be consistent with spatial/temporal features evident in the data, except for an artificial component resulting from latency jitter. Subject scores on this component are shown to reflect peak latencies in the data, suggesting a new aspect to statistical analyses based on subject scores. In general, the results support the conclusion that the TC model is a promising alternative to principal components for data reduction and analysis of MEP's.

Brain Mapping↗

Promoter-proximal pausing by RNA polymerase II in vitro: transcripts shorter than 20 nucleotides are not capped.

We have synthesized RNA from cloned adenovirus 2 late promoter DNA in an in vitro transcription extract under UTP-limiting conditions. Under these circumstances, almost all of the alpha-amanitin-sensitive RNA produced is shorter than 20 nucleotides; most of these short transcripts are present in four species, 6, 7, 13, and 17 nucleotides. These short RNAs are initiated at the adenovirus 2 promoter, as judged by partial sequence analysis and by the abolition of their synthesis upon cleavage of the template DNA at sites which also abolish the production of full-length transcripts. All of the short transcripts can be chased, with excess UTP, into 197-base run-off transcripts; thus, these RNAs are precursors of full-length transcripts and not synthetic "dead ends." Significantly, none of these short RNAs is capped or 2'-O-methylated. However, 79-base run-off transcripts synthesized from this promoter with nonlimiting NTP levels are fully capped.

Base Sequence↗

Nutritional regulation of degradation of aspartate transcarbamylase and of bulk protein in exponentially growing Bacillus subtilis cells.

The rate of degradation of aspartate transcarbamylase in exponentially growing Bacillus subtilis cells was determined by measurement of enzyme activity after the addition of uridine to repress further enzyme synthesis and by specific immunoprecipitation of the enzyme from cells grown in the presence of [3H]leucine. Aspartate transcarbamylase was degraded with a half-life of about 1.5 h in cells growing on a glucose-salts medium with NH4+ ions as the sole source of nitrogen. Replacement of NH4+ in this medium with a combination of the amino acids aspartate, glutamate, isoleucine, proline, and threonine reduced the degradation rate to an undetectable level. Various other amino acids and amino acid mixtures had smaller effects on the rate of degradation. The carbon source also influenced the degradation rate, but to a smaller extent than the nitrogen source. The effects of these nutritional variables on the rate of bulk protein turnover in growing cells were generally similar to their effects on degradation of aspartate transcarbamylase. Since the degradation of aspartate transcarbamylase has been shown to be 10 to 20 times faster than bulk protein turnover, the results suggest that a substantial portion of protein turnover in growing cells represents regulable, rapid degradation of a number of normal proteins, of which aspartate transcarbamylase is an example.

Amino Acids↗