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PubMed1993

High rates of survival and fertilization of mouse and hamster oocytes after vitrification in dimethylsulphoxide.

A high proportion (> 70%) of mouse and hamster oocytes exposed for 3-5 min to 1.5 M dimethylsulfoxide (DMSO) and washed briefly in 3.9 M DMSO before vitrification in 6.0 M DMSO appeared morphologically normal on recovery. Significantly fewer (< 46%) mouse oocytes appeared normal when the time of exposure to 1.5 M DMSO was reduced to 1 min or less. The rate of fertilization in vitro of vitrified oocytes was reduced compared to the rate for untreated controls (mouse: 79% vs. 94%; hamster: 73% vs. 87%). After removal of the zona pellucida, fertilization was similar in vitrified and control hamster oocytes inseminated with hamster (> 90%) or human (21% vs. 23%) sperm. Sperm nuclear decondensation and pronuclear formation appeared to be delayed in the cytoplasm of vitrified hamster oocytes. Seventy-nine percent of 2-cell-stage mouse embryos derived from vitrified oocytes implanted after transfer to pseudopregnant recipients, but only 40% developed to normal fetuses compared to 61% of controls. The reason for this high rate of postimplantation loss is unknown.

Animals↗
PubMed1995

Cytogenetical analysis and developmental potential of vitrified mouse oocytes.

Mature mouse oocytes were cryopreserved by vitrification in 6 M dimethyl sulfoxide (VS). After warming they were either artificially activated with strontium (Sr2+), and the incidence of chromosome non-disjunction was assessed at first cleavage metaphase; or they were fertilized in vitro, and postimplantation survival was examined at Day 15 of gestation. Similar proportions of vitrified and freshly collected oocytes were activated with Sr2+ (75% and 82%, respectively). The majority of activated oocytes extruded the second polar body and formed a single pronucleus ( > 90%). When the exposure time to VS was increased from 90 to 110 sec without cooling, a significant proportion of activated oocytes arrested at the pronuclear stage (30%), and chromosome condensation did not occur. The frequency of aneuploidy in vitrified and control oocytes was similar, but when exposure to VS without cooling was extended, aneuploidy and second polar body retention were significantly higher than those of controls (p < 0.05). The rates of fertilization of vitrified (85%) and control oocytes (92%) did not differ. After transfer, similar proportions of vitrified and control embryos implanted (68-80%) and formed normal fetuses (38-49%). We conclude that vitrification in 6 M dimethyl sulfoxide is a simple and safe procedure for the preservation of mouse oocytes provided that the time of exposure to the cryoprotectant is carefully controlled.

Aneuploidy↗
PubMed1989

Comparison of oral transmucosal fentanyl citrate and an oral solution of meperidine, diazepam, and atropine for premedication in children.

The safety and efficacy of premedication with oral transmucosal fentanyl citrate (OTFC) was compared with that of an orally administered solution of meperidine, diazepam, and atropine and no premedication in 59 children about to undergo elective operations. The patients were randomly assigned to receive no premedication (n = 19); 0.25 ml/kg of the oral solution (containing meperidine, 1.5 mg/kg, diazepam, 0.2 mg/kg, and atropine, 0.02 mg/kg, n = 20); or OTFC (15-20 micrograms/kg, n = 20). Children had activity (sedation) and anxiety scores, vital signs (including systolic and diastolic arterial blood pressures and heart and respiratory rates) and pulse oximetry determined oxygen saturation measured before and at 10-min intervals after premedication until they were taken to the operating room. Quality of induction and recovery was evaluated using scoring schedules; recovery times were measured and side effects noted. OTFC was readily accepted and provided significant reductions in preoperative activity (sedation) and anxiety starting after 30 min. After OTFC, sedation and anxiolysis were significantly greater than in children having no premedication but similar to children having the oral solution for premedication. Vital signs and oxygen saturations remained unchanged preoperatively in all groups. Induction and recovery evaluations and recovery times were similar in the three groups, although children having OTFC had the lowest requirements for narcotics in the recovery room. OTFC caused an 80% incidence of mild preoperative facial pruritus and a higher overall incidence of postoperative vomiting (37%) than premedication with the oral solution (5%) or no premedication (18%).(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMedDate not supplied

The truth and consequences of enzymatic detergents.

Enzymatic detergents are widely used to clean gastrointestinal endoscopes before high-level disinfection. Although proper use of enzymatic detergents may appear straightforward, common misuses include failure to dilute the enzymatic detergent, overdilution of enzymatic detergent, use of expired enzymatic detergent, inadequate exposure time, and failure to rinse the enzymatic detergent from the instrument being cleaned. Each type of misuse has its own rationale. In these studies, the authors show by high-performance liquid chromatography that the type of enzymatic detergent and the dilution and rinsing of enzymatic detergents affect the amount of residual high-level disinfectant (ortho-phthalaldehyde) left on test segments of flexible endoscope insertion tubes. The authors also qualitatively demonstrate that proteinaceous material, which is stained a dark color by ortho-phthalaldehyde (OPA), remains on colonoscopes that have been cleaned with improperly diluted or rinsed enzymatic detergents. These findings emphasize the importance of diluting and using enzymatic detergents exactly as directed by their manufacturers to reduce bioburden and residual amounts of high-level disinfectant on flexible endoscopes.

Disinfectants↗
PubMed2002

Improving medical students' comfort with, and skill in handling, gender-related inquiries during the residency selection process.

OBJECTIVE: Previous work has identified that medical students experience instances of gender discrimination and sexual harassment during the residency selection process (i.e., "audition" elective rotations, residency interviews, and career counseling).(1,2) Examples of such instances include male students who report being counseled away from obstetrics-gynecology and a female student who reported being asked about her sexual orientation because of an expressed interest in breast disease. Students also report confusion about what constitutes an appropriate question or inappropriate gender-related comments. A common example is questioning about plans for child care during residency. We developed an education intervention focused on gender- and family-related issues that may arise during the residency selection process in an attempt to improve our students' ability to (1) recognize appropriate and inappropriate questions and situations and (2) handle such situations in a professional manner. DESCRIPTION: Our educational intervention is a voluntary, intense, active learning situation composed of four parts. First, data outlining the problem-including personal stories of students-are presented. Second, a faculty member with dual experience as a residency training director and dean for student affairs presents examples of appropriate and inappropriate questions (both types of questions can be uncomfortable if not contemplated in advance), potential ways to approach these questions, and a thoughtful approach for the student to take when integrating his or her experiences during the residency selection process with ultimate decisions about career and ranked programs. During the third part of the workshop, students draw two cases of 15 potential situations from a hat. They work in pairs, first to identify whether the case outlines an appropriate or inappropriate situation and second to develop and practice their responses. The session concludes with open discussion about the various scenarios and responses. DISCUSSION: This workshop has been well received by our medical students. In post-workshop surveys, students universally reported that the session was useful. Ninety percent of attendees felt better able to recognize appropriate and inappropriate questions and situations during the residency selection process. Seventy-seven percent of students reported that they felt better able to handle inappropriate situations during the residency selection process in a professional manner. Student comments reflected great satisfaction with the interactive nature of the exercise, interest in expanding the amount of time dedicated to the workshop, and a desire for clear rules about what can/cannot be said. Faculty offering the workshop reported satisfaction doing the workshop and noted that the greatest challenge in presenting the material is balancing the discussion about reasonable responses to inappropriate behaviors with the students' interest in performing well on all interviews in order to maximize their options. Interest in the program has resulted in the authors' being invited to share the workshop at an upcoming GME retreat for the institution's program directors. Based on these results we plan to continue the workshop in an expanded version. In addition, we plan to use this format of the workshop as a springboard for similar sessions for medical students interested in confronting issues of discrimination and harassment in the educational setting.

Female↗
PubMed1996

Content of brain aluminum is not elevated in Alzheimer disease.

Several studies have reported that the bulk aluminum (Al) concentration is increased in the brain in Alzheimer disease (AD), while other studies have failed to demonstrate an increase. Most of these investigations have had one or more methodological deficiencies, including lack of adequate neuropathological assessment; failure to age-match the control samples; small sample sizes, lacking statistical power; and geographical heterogeneity in the AD and control populations. The present population-based study of 92 clinically and histopathologically diagnosed AD patients and normal elderly nursing home residents was designed to avoid these potential biases. When a subsample of AD cases with the most severe brain pathology was compared with controls having no or minimal pathology, no statistically significant differences were found in the bulk aluminum concentration measured by graphite furnace atomic absorption spectrometry in frontal cortex (1.8 +/- 0.7 vs. 1.7 +/- 0.7 micrograms/g dry wt), temporal cortex (1.4 +/- 0.3 vs. 1.5 +/- 0.5 micrograms/g dry wt), liver (2.0 +/- 1.3 vs. 2.0 +/- 1.2 micrograms/g dry wt), or head of femur (2.4 +/- 1.6 vs. 2.2 +/- 1.0 micrograms/g ash wt). Within the whole series of 92 cases, there was no difference in the bulk aluminum concentration of the frontal cortex between individuals diagnosed as definite, probable, and possible cases of AD using the CERAD (Consortium to Establish a Registry for Alzheimer's Disease) criteria. The density of senile plaques and neurofibrillary tangles in frontal and temporal cortex showed no correlation with the bulk aluminum concentration. Logistic regression analyses, which controlled for age and sex, did not influence outcome for any of the comparisons. The data show conclusively that in AD, bulk aluminum concentration is not increased in two cortical brain regions that are selectively vulnerable to the neuropathological changes associated with this disorder.

Aged↗
PubMed2000

Association analysis of beta2 adrenoceptor polymorphisms with hypertension in a Black African population.

OBJECTIVE: To determine whether or not beta2 adrenoceptor polymorphism is a risk factor for the development of hypertension in a Black South African population. BACKGROUND: Attenuated vasodilator responses to endogenous catecholamines may contribute to the aetiology of hypertension. Downregulation of beta2 adrenoreceptors (beta2AR) following stimulation with agonists is determined in part by variation at the beta2AR gene locus. The Glu27 beta2AR genotype results in attenuated downregulation compared with the wild-type Gln27 receptor, whereas Gly16 exhibits enhanced down-regulation compared to Arg16. Possible racial differences in the prevalence of the beta2AR polymorphisms may be an explanation for the blunted responses to isoprenaline and the increased prevalence of hypertension in Black African populations. METHODS: One hundred and ninety-two unrelated hypertensives and 123 normotensives of Black South African origin were studied. Hypertensives were recruited from hospital hypertension clinics in the province of Gauteng and if on treatment, had a 2-4 week washout period before 24-h ambulatory blood pressure assessment Normotensive controls were recruited from the same community. RESULTS: There was no significant association between either the Arg-Gly16 polymorphism or the Gln-Glu27 polymorphism and hypertension status. Furthermore, in the hypertensives, no significant association was seen between beta2AR genotype at either site and clinical blood pressure, 24-h blood pressure or left ventricular mass. A significant association was seen between Arg16 homozygotes and lower body mass index in hypertensives (P = 0.007) although this was not a primary end point. Interestingly, the Glu27 polymorphism was much rarer in this population (allelic frequency 17%) compared to a Caucasian population. CONCLUSION: These data suggest that beta2AR polymorphism is not a risk factor for hypertension per se in this defined population. The possibility that the decreased prevalence of Glu27 in black South African populations explains blunted vasodilator responses to isoprenaline requires further study.

Adrenergic beta-Agonists↗
PubMed1987

Sweets, chocolate, and atypical depressive traits.

An original questionnaire, the Foods and Moods Inventory (FMI) was used to investigate appetite for sweets and chocolate and its relationship to dysphoric mood. The FMI was administered to a group of subjects with an identified interest in chocolate (chocolate group, N = 73), a comparison sample (comparison group, N = 172), and a sample of former alcoholics (N = 22). Those who reported "self-medicating" with sweets or chocolate were more likely to have personality traits associated with hysteroid dysphoria, an atypical depressive syndrome. In addition, the tendency to eat compulsively, in general, and appetite for sweets and chocolate, in particular, were significantly greater among women.

Adult↗
PubMed1982

Loperamide modifies Escherichia coli, heat-stable enterotoxin-induced intestinal secretion.

We have shown previously that loperamide, an opiate analogue, inhibits cholera-toxin- and prostaglandin E2-induced secretion in the rat small intestine. In these studies loperamide modified secretion induced by partially purified Escherichia coli, heat-stable enterotoxin in infant mice. The drug was effective whether administered before or after established secretion. These experiments provide further suggestive evidence that loperamide has a broad spectrum of antisecretory activity.

Animals↗
PubMed1983

Plasmid-mediated adhesion in enteropathogenic Escherichia coli.

A survey of classical serotype enteropathogenic Escherichia coli has been made with respect to their plasmid profile and ability to adhere to HEp-2 cells. Thirty-one of the 32 strains examined contained a 50-70 Md plasmid, and many exhibited HEp-2 adherence. Strain E2348 (0127:H6), which causes diarrhea in volunteers and is HEp-2-adhesive, was chosen for further study. The large 55 Md plasmid in E2348, pMAR2, has been marked with a transposon coding for ampicillin resistance. E2348 that has been cured of pMAR2 loses the ability to adhere to HEp-2 cells, while HB101, a nonadherent E. coli K12, acquires HEp-2 adhesiveness after gaining this plasmid. Plasmid presence was also shown to correlate with in vivo adhesion to intestine, using the colostrum-deprived piglet model.

Animals↗
PubMed1985

Widespread cutaneous Crohn's disease presenting in childhood.

Cutaneous lesions containing Crohn's granulomata are rare and tend to occur in submammary or abdominal wall skin folds when they are to be found (1). Crohn's disease of the skin in nonflexural locations has been reported (2,3) although not in children. We report a 5-year-old girl with extensive colonic Crohn's disease, who at presentation was noted to have a generalised erythematous papular rash which histologically showed typical noncaseating granulomata.

Cecum↗
PubMedDate not supplied

An electron microscopic investigation of time-related changes in the intestine of neonatal mice infected with murine rotavirus.

Seven-day-old mice were infected orally with murine rotavirus (EDIM) and regions of the gut examined at 24 h intervals up to 7 days by electron microscopy. Structural changes were correlated with data on viral antigen production, thymidine kinase activity, and clinical signs of diarrhea. No pathological changes were detected in the colon. Infection and structural damage were confined to the small intestine, with middle regions showing the most pronounced changes. Constriction of villus bases, edema of the lamina propria, and vacuolation of enterocytes occurred at 24 h postinfection (PI), i.e., before evidence of major virus replication. Transient villus atrophy occurred at 48 h PI. Recovery of villus length was evident by 72 h PI accompanied by evidence of marked enterocyte replication at villus bases. Many enterocytes were damaged with little evidence for the presence of virus particles. By 96 h PI, villi had almost recovered from infection although some enterocytes were still damaged; no virus particles were detected in these cells. A second phase of villus damage and edema of the lamina propria occurred at 120 h PI; the pathology resembled that at 24-48 h PI. By 144 to 168 h PI, recovery of the mucosa from infection was virtually complete. We suggest that many of the pathological features following rotavirus infection result from rotavirus-induced ischemia of villi and that diarrhea results from malabsorption of fluid by damaged villi and hypersecretion of ions released from increased numbers of dividing cells at villus-crypt borders.

Animals↗
PubMedDate not supplied

Intestinal enzyme profiles in normal and rotavirus-infected mice.

To investigate further the pathophysiology of rotavirus-induced diarrhea, changes in specific activities of eight relevant intestinal enzymes [alkaline phosphatase, thymidine kinase, lactase, maltase, sucrase, Na+,K+-adenosine triphosphatase (ATPase), adenylate and guanylate cyclases] were measured following infection of suckling mice with murine rotavirus (epizootic diarrhea of infant mouse strain) and compared with age-matched control mice. The concentration of lactose within the lumen of the gastrointestinal tract during infection was also measured. During the course of infection, activities of alkaline phosphatase and lactase decreased, whilst the activity of thymidine kinase increased. Precocious maturation profiles of sucrase and maltase enzymes were observed. No significant changes were detected in the activities of Na+,K+-ATPase or the adenylate and guanylate cyclases. These results are discussed in relation to existing and novel hypotheses on the pathogenesis of rotavirus-induced diarrhea.

Alkaline Phosphatase↗
PubMed1990

An in vitro model to study aspects of the pathophysiology of murine rotavirus-induced diarrhoea.

An in vitro system is described and validated for studying transport of solutes and water in both uninfected and rotavirus-infected neonatal mouse intestine. Control intestine exhibited stable water absorption for periods of up to 40 min. Water absorption was temperature-dependent. Na-dependent, and enhanced by glucose-containing perfusion solutions. Theophylline induced net secretion of water by control intestinal tissue. Water transport by rotavirus-infected lower small intestine was significantly depressed as compared to control levels, and rotavirus-infected middle small intestine exhibited net secretion of water. Upper small intestine and colon from infected animals did not differ significantly from control tissues in their ability to transport water. Water secretion by infected middle small intestine was reversed to absorption by glucose-containing solutions.

Animals↗
PubMed1990

X-ray microanalysis of rotavirus-infected mouse intestine: a new concept of diarrhoeal secretion.

Neonatal mice were infected at 7 days of age with rotavirus [epizootic diarrhea of infant mice (EDIM) virus] and killed at 24-h intervals postinfection (PI). Cytoplasmic concentrations of Na, Mg, P, S, Cl, K, and Ca intestinal epithelial cells from infected and age-matched control animals were measured by x-ray microanalysis. In villus tip cells, Ca concentration increased at 24-96 h PI; Na concentration increased at 24-72 h PI; Ca and Na concentrations were near normal by 168 h PI. K concentration decreased 24-72 h PI, and Cl concentration decreased 48-96 h PI. In crypt cells, changes were observed without a discernible pattern: at 96 h PI, Na, Mg, S, and Cl concentrations increased and K concentration decreased; at 120 h PI, the concentrations of all elements except Na and Ca increased. In villus base cells, the mean concentrations of all elements except Ca peaked at 48-72 h PI and at 120 h PI. Na and Cl concentrations increased dramatically in some cells from 48 h PI onward. All the above concentration values were obtained from freeze-dried specimens and expressed in millimoles per kilogram of dry weight. Conversion of a limited number of data, pertaining to villus base cells, from dry weight to wet weight was possible. This conversion revealed that villus base cells in infected animals were more hydrated than corresponding cells from control animals. Also, the Na and Cl concentrations in mmol/kg H2O were significantly higher in villus base cells from infected animals than in those from corresponding controls: 137 +/- 7 versus 38 +/- 4 (Na) and 121 +/- 5 versus 89 +/- 6 (Cl). Wet weight concentrations of other elements were either the same (Mg) or lower (P, S, and K) after infection with virus. From these studies, a new concept of the pathophysiology of rotavirus-induced diarrhoeal secretion is proposed: stimulation of villus base cells to rapid cell division is accompanied by transient accumulation of Na and Cl; excess NaCl is secreted into the lumen, which is the driving force for fluid loss.

Animals↗