Building comparative gene expression databases for the mouse preimplantation embryo using a pipeline approach to UniGene.

To understand early mammalian development there is a need to compare profiles of gene expression from different stages of the preimplantation mouse embryo. We describe here a method that uses gene expression data held in the UniGene database of the National Institutes of Health (NIH). The full mouse UniGene database (build #151) contains 43,104 gene clusters generated from approximately 4.1 million sequences. The Expressed Sequence Tags (EST) used to build UniGene are derived from cDNA libraries that are archived separately in the database of Expressed Sequence Tags (dbEST) database, with their own catalogue numbers. The mouse dbEST database contains 32 non-normalized dbEST libraries constructed from preimplantation stages (unfertilized oocyte, fertilized oocyte, 2-, 4-, 8- and 16-cell embryo and blastocyst). These libraries contain 219,852 EST sequences mapping to 15,731 UniGene clusters. We have developed a computational pipeline approach that imports and aggregates inventories of gene expression contained in these dbEST libraries. It uses these data to build an annotated web-based database of preimplantation gene expression with an in-built capacity for comparison of expression profiles. Comparison of gene expression profiles obtained for each developmental stage show statistically significant changes in gene expression during preimplantation development. These in silico-generated profiles were validated using RT-PCR.

Using expressed sequence tag databases to identify ovarian genes of interest.

GenBank contains 4879 expressed sequence tags (EST) derived from four non-normalized human ovarian cDNA libraries. Of these EST, 2646 are contributors to UniGene clusters and have UniGene numbers. The EST map to 1206 distinct UniGenes. A gene expression profile was established for the human ovary by identifying the abundance of each UniGene cluster and its corresponding annotation. The most highly expressed transcripts were for proteins associated with protein synthesis (ribosomal proteins, elongation factors, thymosins, etc.). However, there are also transcripts for genes of unknown function that are ovary-specific. This ovarian gene expression profile provides useful data for the design of DNA microarrays targeted at ovarian function and highlights novel sequences that warrant further investigation.

A case of multiple splenic abscesses managed non-operatively.

We report the case of a previously fit 22-year-old man who developed multiple splenic abscesses of unclear aetiology and was successfully treated with percutaneous drainage of the largest abscess and antibiotic therapy.

Metastasising dermatofibroma or dermatofibroma-like dermatofibrosarcoma protuberans?

Dermatofibroma is a relatively common cutaneous fibrohistiocytic tumour with a marked tendency to recur locally. It is almost invariably benign. In a review of the world literature there was one report of two patients with dermatofibroma with histologically documented metastases. In both of these there was local recurrence and subsequent metastasis to lymph nodes and haematogenous spread to the lungs. We present a case of cellular dermatofibroma of the lower limb with local recurrence and metastasis to the abdominal wall, posterior chest wall and both lungs 30 months after excision of the primary lesion. The clinical and pathological features of the case are presented to emphasis the need for follow up of patients with dermatofibroma which present any unusual histological features.

The Scottish Royal Colleges: a prospective survey of continuing medical education undertaken by their examiners.

OBJECTIVE: To measure the amount and nature of Continuing Medical Education (CME) activity undertaken by the examiners of the three Scottish Royal Colleges. The data obtained to be compared with recommendations for formal CME participation published by the medical Royal Colleges and Faculties. DESIGN: All examiners in active clinical practice prospectively surveyed over a 12 month period by completing a monthly return from a specially designed loose-leaf diary. RESULTS: 75% of the examiners who submitted all 12 monthly diary pages completed at least 100 hours CME during the year while 95% achieved 50 hours or more. This time was distributed approximately equally between hospital-based activities and activities for which study leave might have been taken. Although the mean total number of hours of CME undertaken by consultants based in district general hospitals was also over 100 hours in the year, this was significantly lower than the figure for teaching hospital consultants. The type of CME activity was described as general (16%), specialty (49%) and subspecialty (35%). The examiners surveyed found that specialist society and international specialist meetings were perceived to be the most valuable of those attended. CONCLUSIONS: The current recommendations for implementing formal CME systems for consultants are in line with the current activity levels of this group and should safeguard existing educational activity rather than impose new standards. This study suggests that the profession should not be unduly anxious about the introduction of a structured CME system although it is likely that some form of obligatory scheme will be required to achieve a more uniform response.

Evidence of electromechanical dissociation of the internal anal sphincter in idiopathic fecal incontinence.

PURPOSE: This study was designed to evaluate the relationship between internal sphincter electromyographic frequency and ambulatory anal pressures in order to clarify the pathophysiology of internal anal sphincter dysfunction in fecal incontinence. METHODS: Seventy-two patients of median age 55 years (range, 24-75; 63 females) with neurogenic fecal incontinence and 33 normal subjects of median age 48.5 years (range, 25-74; 21 females) underwent fine-wire anal sphincter electromyography and anal manometry. RESULTS: The median internal anal sphincter electromyographic frequency was incontinent 0.25 Hz (0.2-0.34) and the control was 0.44 Hz (0.36-0.55; P < 0.03). Ambulatory resting pressures were incontinent median 54 cm of H2O (34-68 cm of H2O) and control 94 cm of H2O (72-102; P < 0.01). Internal sphincter electromyographic frequency correlated with anal resting pressures in both groups (P < 0.002). Internal sphincter electromyographic silence not attributable to electrode movement or the rectoanal inhibitory reflex, lasting 0.5 to 4 minutes occurred in all but two of the incontinent patients. The anal pressure during this period did not significantly change (P > 0.1). No recruitment of the external sphincter or puborectalis was noted during these episodes. Such electromechanical dissociation was not seen in the control group. The frequency of transient internal sphincter relaxation was 4 (ranges 2-6) per hour in controls and 8 (ranges, 6-12) per hour in incontinent patients (P < 0.01). Rectal pressures did not exceed midanal pressures in any of the controls but did in all of the incontinent patients on at least one occasion per hour in the incontinent group. CONCLUSION: Internal anal sphincter activity exhibits electromechanical dissociation and relaxes abnormally in incontinent patients.

Rectoanal inhibition and incontinence in patients with rectal prolapse.

Thirty-five patients with complete rectal prolapse, 32 with neurogenic faecal incontinence and 33 controls underwent ambulatory recording using a computerized anal electromyographic and anorectal manometry system. Median resting anal pressures were 34 cmH2O in patients with prolapse, 51 cmH2O in those with neurogenic faecal incontinence and 94 cmH2O in controls. Median basal rectal pressures were 18, 21 and 21 cmH2O respectively. High-pressure rectal waves of median amplitude 71 cmH2O lasting 30-150 s and associated with inhibition of the electromyographic activity of the internal and sphincter and a fall in anal pressures were seen in all patients with prolapse but not in controls or those with neurogenic incontinence. These waves were abolished following successful resection rectopexy. Recovery of continence occurs by abolition of high-pressure rectal waves, which produce maximal inhibition of sphincter activity before operation.

Sustained internal sphincter hypertonia in patients with chronic anal fissure.

PURPOSE: This study was designed to determine whether functional variations of internal sphincter activity occur in order to differentiate between patients with anal fissures from those with hemorrhoids. METHODS: Thirty patients with chronic anal fissure (median age, 28 years; 12 females), 22 patients with hemorrhoids (median age, 37 years; 7 females), and 33 control volunteers (median age, 48.5 years; 21 females) underwent ambulatory anal sphincter fine-needle electromyography and anorectal manometry. RESULTS: The median internal sphincter electromyography frequency was similar: fissure group, 0.49 Hz; hemorrhoid group, 0.46 Hz (P > 0.05), and control group, 0.44 Hz (P > 0.05). Median anal resting pressures were similar in the fissure group (132 cm. H2O) and the hemorrhoids group (116 cm of H2O) (P > 0.05), but significantly greater than those in the control group (94 cm. H2O) (P < 0.05). The median number of transient relaxations of the internal and sphincter with an associated rise in rectal pressure and fall in anal pressure was 1 (range, 0-4) per hour in the fissure group, 6 (range, 4-7) per hour in the hemorrhoid group, and 4 range, 3-6) per hour in the control group. Six patients with fissures were reassessed following lateral internal sphincterotomy. Median and pressure was 102 cm of H2O (P > 0.1 vs. controls) and the number of internal sphincter relaxations increased to 4 per hour (P < 0.01 vs. preoperative number). CONCLUSIONS: Internal anal sphincter relaxation occurs on fewer occasions in patients with chronic anal fissures that have failed to heal in comparison to patients with hemorrhoids and normal controls. This evidence further supports the hypothesis that internal sphincter hypertonia may be relevant to the pathogenesis of this disorder.

Restoration of continence following rectopexy for rectal prolapse and recovery of the internal anal sphincter electromyogram.

Twenty-two patients with full-thickness rectal prolapse underwent ambulatory fine wire electromyography of the internal and sphincter (IAS), external and sphincter and puborectalis, together with anorectal manometry, using a computerized system. Examinations were performed both before and 3 to 4 months after rectopexy. The median (interquartile range (i.q.r.)) preoperative IAS electromyogram (EMG) frequency was 0.18 (0.05-0.31) Hz and the median (i.q.r.) preoperative resting anal pressure was 28 (15-64) cmH2O. An improvement in the IAS EMG frequency, median (i.q.r.) 0.29 (0.19-0.38) Hz (P less than 0.03), and resting anal pressure, median (i.q.r.) 41 (20-72) cmH2O (P less than 0.05), was recorded after operation, but these variables remained significantly lower than those found in normal controls: median (i.q.r.) IAS EMG frequency 0.44 (0.36-0.48) Hz and median (i.q.r.) resting anal pressure 92 (74-98) cmH2O. We suggest that repair of the prolapse allows the IAS to recover by removing the cause of persistent rectoanal inhibition.

Combined treatment with BCG and chemotherapy for metastatic gastrointestinal cancer.

The survival of patients who have cancers of the gastrointestinal tract is poor even after surgical resection of all gross disease. In an initial consecutive series of patients with metastatic or recurrent cancers, an attempt to improve survival with combined nonspecific immunostimulation by the intraperitoneal administration of BCG followed by oral BCG cycled with chemotherapy was studied. The results suggested that a population of patients with gastric and colonic cancers showed improved survival with this treatment. A randomized trial of combined chemoimmunotherapy versus chemotherapy alone was begun on November 1, 1974, for all cancers of the stomach and pancreas and all colorectal cancers beyond stage C by Dukes' classification. The initial results with patients entered by December 31, 1975, and followed until March 1, 1976, are reported.

Adjuvant immunostimulation in malignant melanoma with oral Bacille Calmette-Guérin.

Results of the administration of oral bacille Calmette-Guérin (BCG) as an adjunct to standard treatment in 62 patients with malignant melanoma indicate that this drug is of value in preventing distant spread of the disease when it is limited to one region and in its early stages. BCG increases survival in patients with visceral metastases. In those in whom these metastases are surgically resectable, it inhibits the development of further metastases. Oral BCG treatment does not affect the course of the disease in patients with massive hepatic or intracranial metastases.

Immunostimulation with intraperitoneally administered bacille Calmette Guérin for advanced malignant tumors of the gastrointestinal tract.

Thirty-five patients with advanced malignant tumors of the stomach, pancreas, biliary tract or colon were treated intraperitoneally with bacille Calmette Guérin and, subsequently, were given orally booster doses of bacille Calmette Guérin and one of two schedules of cytotoxic drugs. Patients with all types of tumors responded, but this was most marked in patients with metastases to the liver from primary carcinoma of the colon.

Canine small bowel transplantation. A study of the immunological responses.

A canine small bowel allograft model was used to determine the effects of radiation to the graft in modifying the immunological effects of the passenger leukocytes. When untreated allografts were transplanted, death of the recipient animals occurred at a mean of nine days. The allograft was well-preserved and showed no signs of rejection. The reasons for attributing death to graft-versus-host (GVH) disease are discussed. When allografts were treated with 150 rads prior to transplantation, allograft rejection occurred, with death of the recipient animals at a mean of 9.2 days. This was the only group in which cell-mediated immunity developed. When allografts were treated with 50 rads, prolonged survival of the recipients to a mean of 28 days was noted. It is postulated that in this group a balance was struck between the allograft rejection reaction and GVH disease, with prolongation of allograft survival.

Oral bacille Calmette Guérin immunostimulation in malignant melanoma.

Forty-seven patients representing all stages of malignant melanoma have been treated orally with bacille Calmette Guérin as an adjunct to standard treatment regimens. This is a preliminary study, but it appears that orally administered bacille Calmette Guérin in localized disease may delay the development of local recurrence and distant spread. In patients with disseminated melanoma but without intracranial metastases, the survival time is increased from 4.0 to 12.7 months. It has no effect on the progress of disease in patients with intracranial metastases or in those with hepatic metastases from intraocular melanoma.