Adenocarcinomain Situof the Uterine Cervix: Management and Outcome

Objective: To retrospectively review the management of adenocarcinomain situof the uterine cervix, to determine the outcome of conization versus hysterectomy, and to compare the results achieved by different methods of conization. Methods: We performed a retrospective pathology and chart review of 46 patients with cervical adenocarcinomain situfrom January 1980 to October 1994. Results: Nine patients were managed during the first half of the study period and 37 were managed in the second half. The mean age of patients was 38.4 years (range 25–72). Forty-five of 46 patients were diagnosed as a result of an abnormal Pap smear, although only 19 smears indicated adenocarcinomain situor other glandular abnormalities. Cold knife conization resulted in a 33% rate of positive margins for adenocarcinomain situcompared to 50% for large loop excision of the transformation zone (LLETZ). Among 24 conservatively managed patients with negative conization margins, there have been 2 (8.3%) recurrences of adenocarcinomain situ.Among patients not undergoing hysterectomy as definitive treatment, 1 of 18 (6%) patients undergoing cold knife conization recurred, compared to 4 of 14 (29%) managed with LLETZ, despite a 63.4-month shorter mean follow-up interval for the LLETZ patients. Conclusions: Cold knife conization is associated with a lower rate of recurrence of cervical adenocarcinomain situcompared to LLETZ. We recommend cold knife conization for patients who are not treated with hysterectomy.     

Melatonin responses to clonidine and yohimbine challenges.

Melatonin (MT) release from the pineal gland has been used as a marker for central noradrenergic function in major depression. Norepinephrine acts at both alpha and beta adrenergic receptors on the pinealocyte membrane to mediate nocturnal MT release, but in humans the contribution of each receptor class is unclear. In order to explore the effect of alpha 2 receptors on MT release, 10 female subjects were given oral challenges, in separate placebo-controlled trials, of either 10.8 mg of yohimbine, an alpha 2 adrenergic antagonist, or clonidine, a partial alpha 2 adrenergic agonist, at doses of either 200 micrograms or 300 micrograms. Post-challenge serum melatonin was measured from 18:00 h to 22:00 h in both studies as was urinary 6-sulphatoxy-melatonin (aMT6s), the major metabolite of MT (from 18:00 h to 22:00 h, and from 22:00 h to 10:00 h). Growth hormone (GH) was also assayed following the clonidine challenge, and blood pressure, pulse rate, and side effects were monitored after both challenges. Neither yohimbine nor clonidine significantly altered nocturnal serum MT levels compared to placebo. However, there was a significant increase in urinary aMT6s between 18:00 h and 22:00 h following yohimbine ingestion. Yohimbine ingestion produced significant rises in pulse rate and the urge to urinate compared to placebo. Both doses of clonidine resulted in a significant reduction in pulse rate, systolic and diastolic blood pressure, and significant increases in drowsiness and other measures of sedation following ingestion. Only clonidine 300 micrograms produced a significant elevation in GH release. This study highlights the limitations of oral neuroendocrine challenge studies.     

Validation of computerized three-dimensional reconstruction of intravascular ultrasound: measurements of absolute luminal diameter and cross-sectional area in ex vivo human coronary arteries

Computer based 3-dimensional reconstruction transforms 2-dimensional intravascular ultrasound images into a longitudinal format facilitating analysis of luminal narrowing. To validate the accuracy of current software in measuring coronary artery diameter and cross-sectional area, in arteries with atherosclerosis, we performed 3-dimensional reconstruction in 10 human pathologic coronary arterial segments of 10-25mm length. Images were obtained using a 4.8 French catheter with pullback speed of 1mm/sec acquired at 3 frames/sec onto VHS tape. The data were digitized and intraluminal 3-dimensional reconstruction performed using a voxel-based program. Pathologic sections were obtained every 3mm, and dimensions were measured with a resolution of 0.01 mm. Maximum, minimum, and 3 other representative diameters were recorded by an observer blinded to the ultrasound diameters. Average histo-pathologic diameters were reported, and specimen cross-sectional area was then calculated. Results: In 53 sections, pathological diameters ranged from 1.4-4.5mm (mean 2.7 +/- 0.68mm) while 3-dimensional reconstructed diameters were 1.9 to 3.8mm (mean 2.6 +/- 0.54mm). Pathologic and ultrasound derived 3-dimensional reconstruction diameters had an excellent correlation (r=0.86, SEE=+/-0.36). Pathology and 3-dimensional reconstruction cross-sectional area also correlated closely (r=0.88, SEE=+/-1.50). Diameters less than 2.0mm were systematically overestimated and diameters greater than 3.5mm underestimated by 3-dimensional reconstruction. Most 3 dimensional reconstruction values were within +/- 10% of pathology, but diverged at each diameter extreme, approaching +/- 20%. Thus, computerized 3-dimensional reconstruction of ultrasound images shows excellent quantification of luminal size in the 2.0-3.5mm range, suggesting important investigative and clinical applications.     

Polymorphisms of the equine major histocompatibility complex class II DRA locus

The full extent of the polymorphism of ELA-DRA in Equidae is not yet known. Given the apparent differences in DRA polymorphisms between Equidae and other species, the aims of this study were to more fully characterize ELA-DRA, determine the extent of gene polymorphism and establish the allele-frequency distribution. An allele reference panel for the second exon of ELA-DRA was established by sequence-based typing of 69 equine DNA samples consisting of various breeds of domestic horse (Equus caballus), together with donkeys (Equus asinus), Grant's zebras (Equus boehmi) and one onager (Equus hemionus). Five of the six previously reported alleles detected using single-strand conformation polymorphism were found: ELA-DRA*0101, ELA-DRA*0201, ELA-DRA*0301, ELA-DRA*0501 (Albright-Fraser DG et al. Polymorphism of DRA among equids. Immunogenetics 1996: 43: 315-7) and ELA-DRA*0601 (GenBank accession number AF5419361). In addition to the previously reported alleles, five novel ELA-DRA alleles were detected within the ELA-DRA allele reference panel. One of these was identified in E. caballus (ELA-DRA*JBH11), one in E. boehmi and E. hemionus (ELA-DRA*JBZ185) and three in E. asinus (ELA-DRA*JBD3, ELA-DRA*JBD17 and ELA-DRA*JBH45). A total of 565 equine DNA samples were screened using reference-strand-mediated conformation analysis, a double-stranded conformation-based mutation detection system that can be used to type existing ELA-DRA alleles and identify new variants. Based on our findings, at least 11 ELA-DRA alleles are now known to exist, and this level of polymorphism at the DRA locus appears to be unique to the genus Equus. Both the previously reported alleles and the new alleles displayed a species-specific distribution.     

An assessment of the delayed hypersensitivity reaction to methylated bovine serum albumin in the mouse and its use in the evaluation of drug effects on cell mediated immune reactions

A delayed hypersensitivity (DH) reaction is induced by a sensitizing intradermal injection of methylated bovine serum albumin (MBSA) into the abdomen of mice and a subsequent challenge injection of MBSA into the hind paw. Paw volume increase is measured by mercury plethysmography. The conditions for sensitization have been investigated. Sensitization with a 0.25% MBSA emulsion resulted in a small but significant swelling of the paw following the challenge injection. The magnitude of the footpad response to the challenge injection was increased if the antigen administered in the sensitizing injection was emulsified with Freund's incomplete adjuvant. Incorporation ofMycobacterium butyricum in the emulsion greatly increased the footpad response if added at doses of 0.05 and 1 mg per animal. A higher dose (4 mg), however, resulted in a lower response. The time course of development of the delayed hypersensitivity reaction has been studied. An 8-day interval between sensitization and challenge resulted in a greater delayed hypersensitivity response than a shorter (3-day) or longer (15-, 21-, 28-day) interval. Cyclophosphamide (250 mg kg^–1) administered 3 days prior to the sensitizing injection of MBSA produced a modest enhancement of the DH reaction.On the basis of these studies a protocol for conducting the DH reaction to MBSA was established and the activity of drugs on processes underlying the sensitization phase of the reaction or processes underlying the elicitation phase of the reaction have been examined. Steroid and immunosuppressant drugs were found to inhibit the DH footpad response when dosed during the sensitization whereas several specific-anti-rheumatic and immunoactive compounds were without effect. Indomethacin and sudoxicam inhibited the DH reaction if dosed during the elicitation of the reaction but other non-steroidal anti-inflammatories tested did not significantly reduce the response. The clinically used anti-rheumatic drugsd-penicillamine and levamisole did not inhibit the elicitation phase of the DH response but niridazole at 100 mg kg^–1 did reduce the inflammatory response.This paper has been presented in part as a poster presentation to the British Pharmacological Society, 4–6 January 1978.     

Effects of long working hours and the night shift on severe sleepiness among workers with 12‐hour shift systems for 5 to 7 consecutive days in the automobile factories of Korea

We investigated the effects of 12‐hour shift work for five to seven consecutive days and overtime on the prevalence of severe sleepiness in the automobile industry in Korea. [Correction added after online publication 28 Nov: Opening sentence of the summary has been rephrased for better clarity.] A total of 288 randomly selected male workers from two automobile factories were selected and investigated using questionnaires and sleep‐wake diaries in South Korea. The prevalence of severe sleepiness at work [i.e. Karolinska Sleepiness Scale (KSS) score of 7 or higher] was modeled using marginal logistic regression and included theoretical risk factors related to working hours and potential confounding factors related to socio‐economic status, work demands, and health behaviors. Factors related to working hours increased the risk for severe sleepiness at the end of the shift in the following order: the night shift [odds ratio (OR): 4.7; 95% confidence interval (CI): 3.6–6.0)], daily overtime (OR: 2.2; 95% CI: 1.7–2.9), weekly overtime (OR: 1.6; 95% CI: 1.0–2.6), and night overtime (OR: 1.6; 95% CI: 0.8–3.0). Long working hours and shift work had a significant interactive effect for severe sleepiness at work. Night shift workers who worked for 12 h or more a day were exposed to a risk of severe sleepiness that was 7.5 times greater than day shift workers who worked less than 11 h. Night shifts and long working hours were the main risk factors for severe sleepiness among automobile factory workers in Korea. Night shifts and long working hours have a high degree of interactive effects resulting in severe sleepiness at work, which highlight the need for immediate measures to address these characteristics among South Korean labor force patterns.     

Sequence variation in the 3'-untranslated region of the dopamine transporter gene and attention-deficit hyperactivity disorder (ADHD).

The dopamine transporter gene (DAT1) has been reported to be associated with attention-deficit hyperactivity disorder (ADHD) in a number of studies [Cook et al. (1995): Am J Human Genet 56(4):9993-998; Gill et al. (1997): Mol Psychiatry 2(4):311-313; Waldman et al. (1998): Am J Human Genet 63(6):1767-1776; Barr et al. (2001): Biol Psychiatry 49(4):333-339; Curran et al. (2001): Mol Psychiatry 6(4):425-428; Chen et al. (2003): Mol Psychiatry 8(4):393-396]. Specifically, the 10-repeat allele of the 40-bp variable number of tandem repeats (VNTR) polymorphism located in the 3' untranslated region (UTR) of the gene has been found to be associated with ADHD. There is evidence from in vitro studies indicating that variability in the repeat number, and sequence variation in the 3'-UTR of the DAT1 gene may influence the level of the dopamine transporter protein [Fuke et al. (2001): Pharmacogenomics J 1(2):152-156; Miller and Madras (2002): Mol Psychiatry 7(1):44-55]. In this study, we investigated whether DNA variation in the DAT1 3'UTR contributed to ADHD by genotyping DNA variants around the VNTR region in a sample of 178 ADHD families. These included a MspI polymorphism (rs27072), a DraI DNA change (T/C) reported to influence DAT1 expression levels, and a BstUI polymorphism (rs3863145) in addition to the VNTR. We also screened the VNTR region by direct resequencing to determine if there was sequence variation within the repeat units that could account for the association. Our results indicate that DAT1 is associated with ADHD in our sample but not with alleles of the VNTR polymorphism. We did not find any variation in the sequence for either the 10- or 9-repeat alleles in the probands screened nor did we observe the reported DraI (T/C) variation. Our results therefore refute the possibility of the reported DraI variation or alleles of the VNTR as the functional variants contributing to the disorder.