Hypertension and its correlates in the Hutterite community of Alberta.

The purpose of this study was to determine the prevalence of hypertension and its correlates in the Hutterite population of Alberta. A total of 811 subjects from 38 Hutterite colonies participated in the survey. A self- and interviewer-administered questionnaire gathered information about past medical history, lifestyle habits, and family history. Blood pressure, blood cholesterol, 2 hour postprandial blood glucose and height and weight were also measured. The prevalence of diastolic blood pressure > or = 90 mm Hg was approximately twice that reported in other Canadian surveys: 60% and 30% for the males and females respectively. Half of the participants found to be hypertensive were unaware of their condition and of those aware, 76% were uncontrolled. There were significant correlations between diastolic blood pressure and Body Mass Index (BMI), cholesterol, age and glucose. This study supports the need for the development of culturally appropriate health promotion programs related to hypertension in this population.


A double-blind placebo controlled trial of medroxyprogesterone acetate and cyproterone acetate with seven pedophiles.

Seven of ten pedophiles in hospital completed a double-blind, placebo-controlled two-dose comparison of medroxyprogesterone acetate and cyproterone acetate. Sequential measures during the 28 week study were: patient self-reports, nurses' observations, phallometry, hormone levels and side-effects. The drugs, which performed equivalently, reduced sexual thoughts and fantasies, the frequency of early morning erections on awakening, the frequency and pleasure of masturbation, and level of sexual frustration. Penile responses were also reduced but to a lesser degree and were more variable. Serum testosterone FSH and LH all declined during drug administration, but by the end of the final placebo phase had essentially returned to (or exceeded) pre-drug values. Our experience suggests that only a minority of pedophiles are likely to accept libido-reducing drugs.


Follow-up of participants in a preschool hearing screening program in child care centres.

As part of its response to the Ontario Ministry of Health recommendation that communication disorders in preschool children be identified, the Hamilton-Wentworth Department of Public Health Services performed hearing screening tests on all preschool children in child care centres. Parents of children who had failed the hearing screening test were recommended to take their child to their family physician for further assessment. Out of 1,844 children tested between September 1990 and February 1991, 35 children failed the pure-tone audiometry test administered by nurses. A telephone follow-up interview determined that 26 children with screening test failures had received some medical follow-up. Eight children had a confirmed hearing problem; of these, five were newly diagnosed with a hearing problem, all resulting from fluid build-up in the middle ear. Three children were treated, with two having improved hearing. Of the two children not treated, one was reported to have improved hearing. As with other recently reported preschool hearing screening programs, this study raises doubts about the usefulness of such programs.


Recidivism among individuals detained under a warrant of the Lieutenant-Governor living in the community.

This paper reports the findings of a follow-up study of individuals on a warrant of the Lieutenant-Governor in community placement. Of the 32 individuals who had 8.5 years on average of community access, 6.3% had subsequent convictions. A comparison is made between the risk to the community posed by those on warrants and individuals in the general population. The results suggest that those on warrants have functioned relatively well in the community.


Women's views of their main health problems.

Though there has been increased emphasis on women's health and on community participation in the development of health policy, 'ordinary women' have seldom been asked about their major health concerns. This paper reports on a survey of a stratified random sample of 356 women in Hamilton. Among their main worries regarding health were various cancers and heart disease. The health problems they had experienced in the previous six months which had bothered them most were stress, arthritis, being overweight, migraines/chronic headaches and tiredness. On the basis of these and similar data presented here, it is argued that such community surveys provide an important source of data. They identify somewhat different priorities than approaches which rely on the opinions of experts and other key informants.


Is there a shared somatic disorder? Communication of non delusional symptoms in Nigeria.

In a setting in which somatic symptoms without obvious organic causes are prevalent and in which doctors do not have very convincing explanations to offer, people in prolonged contact with such patients may appear to develop similar symptoms but claim the original patient as the source of their symptoms. At times, it is the original patient that detects the similarity and raises the question of contagion. Based on a study of four Nigerian patients, the author proposes the possible existence of a shared somatic disorder, comparable in many ways to induced psychotic disorder (DSM-III-R) and reopens the discussion on the whole concept of sharing/communication of psychiatric symptoms.


Schizophrenia and substance abuse: prevalence issues.

A review of studies of the prevalence of substance abuse among schizophrenics suggest that both demographic and environmental factors are strong determinants of the extent of substance abuse in this population as well as the type of substances used. In a mid-sized Canadian city, a sample of schizophrenic patients referred to a dual diagnoses clinic were administered the Addiction Severity Index. With age and gender included as a factor, no significant differences were found in the choice or length of use of most drugs between diagnostic groups, contrary to some other reports from large urban centres. Compared with a population of a similar age, there were fewer regular and more former drinkers among the schizophrenic patients. The choice of other substances by the sample reflected the pattern of use in the population at large. This approach to prevalence provides a more balanced perspective of the substance abuse by schizophrenics. The major targets for prevention remain the alcohol, caffeine and tobacco consumption.


The ratio of diagnosed to undiagnosed diabetes in patients 40 years and older.

A cross-sectional survey of a one in five random sample of apparently healthy adults aged 40 years and older was done in 17 family practice clinics in Newfoundland, in order to pick up previously undiagnosed diabetics, by fasting blood sugar (FBS) measurements. From a sample of 2,087 people, 1,767 were tested. Forty-eight persons had an elevated FBS on the first test, but only 19 had a sustained high FBS when a second test was done. There were 16 previously-known diabetics for each new diabetic found; in this population it was not worth screening healthy persons for diabetes. Testing should be restricted to high-risk people only. As a result of this study, the diabetes prevalence in the 40+ population rose from 12.4% to 13.1%.


Lack of an association between endemic giardiasis and a drinking water source.

Waterborne outbreaks of giardiasis have been documented in many areas of North America associated with contaminated surface water supplies. The Greater Vancouver Water District (GVWD) administers the distribution of surface waters to 1.4 million people. We wished to determine if endemic giardiasis was associated with this water supply. One hundred and eighty cases, an equal number of laboratory (enteric) controls and 94 neighbourhood (friend) controls were interviewed by questionnaire. The cases and controls were similar in age, sex and community of residence. Risk factors for giardiasis included having a child under six in the house and travel, both in B.C. but outside of the GVWD, and in Africa, the Americas, south of the U.S. or Asia. The cases drank 3.6 cups of water per day, the enteric controls 3.5 and the friend controls 3.7. These amounts are similar to those reported ingested in outbreak studies. Water consumption was not a risk for endemic giardiasis.


Review of studies of risk and protective factors for psychopathology in children.

Child psychiatry is increasingly focusing on early identification of children at risk and the importance of prevention. Our review attempts to summarize recent studies regarding risk and protective factors in children and address the areas of difficulty inherent in this type of research. Recommendations for future research study design will be discussed.


The role of stress in interpreting the dexamethasone suppression test.

Stress has been implicated as a major confounding factor in the interpretation of Dexamethasone Suppression Test (DST) results. This study was designed to examine the effects of stress on DST results. Fifty patients with high levels of acute, chronic, and environmental stress participated in the study. Each patient was given a comprehensive psychiatric and psychological assessment, a routine administration of dexamethasone, and blood tests of cortisol values. The results indicate that the three measures of stress do not appear to affect levels of cortisol suppression, however, all three measures of stress predicted depression. As expected, DST cortisol levels were related to depression. Results are discussed in terms of their implications for understanding the associations among stress, depression and DST results.


Comfort. Immigrant Hispanic cancer patients' views.

This study focuses on the meaning of comfort from the perspective of immigrant Hispanic cancer patients undergoing treatment at a large Western medical center. Patients (n = 10), in 30 ethnographical interviews conducted in Spanish by a culturally sensitive interviewer identified characteristics, needs, and sources of comfort. The characteristics of comfort were: feeling integrated, functioning and normalcy, care and nurturing, security and safety, control and "comodo." Comfort needs evolved into 6 categories: nurturing, familiar environment, safety, quality of life, normalcy, and "animo." Providers of comfort, as perceived by these Hispanic patients were: self, family, health care providers, and God. The characteristics of comfort most frequently talked about by the Hispanic participants were feeling integrated and nurtured. Integrated, a seemingly complex characteristic of comfort, reflected a sense of inner peace with oneself that went beyond the physical comfort. Function and normalcy were also perceived to be important aspects of comfort. It is important to note that the Hispanic patients on occasion used culturally relevant terms, such as comodo and animo (neither of which are readily translated into English) to convey their impressions of comfort. The study demonstrates that comfort is important in illness and cancer treatment situations, and that for these Hispanic patients it also had influence from their culture.


Factors associated with rehabilitation in head and neck cancer.

Head and neck cancer patients must adapt to changes in facial structure and dysfunctions associated with breathing, eating, sensing, and movement following treatment. The impact of social support, mode(s) of treatment, and disfigurement and dysfunction on rehabilitation outcomes was studied in a convenience sample of 51 cancer survivors. The Sickness Impact Profile, a 136-item self-report instrument that elicited data in 12 activity of daily living areas was used to measure the dependent variable, rehabilitation outcome. Perceived social support was measured with the Personal Resources Questionnaire (Part 2). Facial disfigurement was measured based on the surgical procedure; nine dysfunctions common to head and neck cancer patients were measured on ladder scales and combined to provide a total dysfunction score. Facial disfigurement was not significantly associated with rehabilitation outcomes, but perceived social support and degree of dysfunction were significantly correlated with rehabilitation. No significant differences in rehabilitation outcomes between modes of treatment were found. Findings suggest a need for further research on rehabilitation strategies which improve outcomes for dysfunctions associated with head and neck cancer.


Gaps and contract: evaluating the diffusion of new information. Part I. A description of the strategy.

Time and fiscal constraints on oncology nurses require that continuing education programs yield demonstrable benefit to the clinician, the institution, and patient outcomes. This article describes "gaps and contract," a strategy that provides nurses with a tool to transfer knowledge from theory to practice, in a measurable form. The framework for this strategy lies within the concept of innovation diffusion. This strategy was used by two universities collaborating to educate 912 oncology health professionals in 38 continuing education programs. Part II of this article will describe the measurement of this strategy.


Gaps and contract: evaluating the diffusion of new information. Part II. The measurement of the strategy.

This article is the second of a two-part series describing "gaps and contract," a strategy that provides nurses with a mechanism to transfer knowledge from theory into practice in a measurable format. Two universities under contract from the Pennsylvania state cancer plan used this strategy to evaluate the effectiveness of a cancer continuing education program. Two hundred seventy-four (274) contracts from one setting and 205 from a second setting are described. Recommendations for using this strategy to measure the efficacy of cancer continuing education programs are included.


Breast self-examination. Knowledge and behavior of Greek female health care professionals working in primary health care centers.

A comparative correlational study was conducted to examine the knowledge and frequency of breast self-examination (BSE) by Greek health care professionals. A total of 268 women working in primary health care centers constituted the sample. The women were 22-64 years of age. Subjects reported low levels of knowledge about BSE and facts related to breast cancer. Multiple regression analysis showed significant relationships between level of knowledge and profession, number of years of health-related education, and frequency of BSE. The sample's frequency of practice of BSE was limited. Only 34.7% of the participants claimed to practice BSE on a monthly basis. Multiple regression analysis revealed statistically significant correlations between BSE behavior and marital status, professional experience with breast cancer, and level of knowledge. The level of knowledge of the participants is considered to be inadequate, taking into account that they work in settings where primary health care is offered. Courses of preventive medicine and health education should be incorporated in medical and nursing curricula.


Difference between nurses' and physicians' approach to denial in oncology.

A psychiatrist and a psychiatric nurse interviewed 33 physicians and 94 nurses at a cancer center about their experience working with oncology patients, including their usual approach to "denial" seen in these patients. Most respondents viewed denial as a useful, nearly universal defense mechanism, potentially capable of interfering with treatment. Physicians described a pattern in which their patients were presented with the facts one time during diagnosis and formulation of a treatment plan, after which denial was allowed and new facts not offered unless the denial was viewed as interfering with the medical intervention. Nurses more often referred to denial as a phase, stressing the importance of honesty in dealing with patients who are prone to deny. They experienced discomfort when patients who were suffering adverse effects of treatment seemed to need greater honesty from their physicians. Discussion of these differences includes the effect of the contexts in which nurses and physicians encounter denial as well as their complementary roles in patient care, and the differential goals and values of the two professions.


Assessment of the autologous bone marrow transplant patient according to Orem's self care model.

Autologous bone marrow transplantation (ABMT) is an increasingly effective treatment option for both hematologic malignancies and solid tumors. The dose-limiting toxicity of bone marrow suppression after intensive chemotherapy and/or radiation therapy can be minimized by reinfusing the patient's stored marrow. However, the ABMT procedure involves a period of profound neutropenia before the reinfused marrow engrafts and it also carries a significant risk of major organ toxicities. Common adverse effects of the procedure include mucositis, pneumonitis, renal failure, and veno-occlusive disease of the liver. In this article, Orem's Self-Care Model is used as a framework for assessing ABMT patients with the goal of recognizing developing complications early.


Hypercholesterolemia in rats with chronic renal insufficiency not aggravated by recombinant human growth hormone.

The lipid metabolic disorders in chronic renal insufficiency (CRI) are related to increased hepatic lipid synthesis, reduced triglyceride removal coupled with insulin insensitivity and impaired lipoprotein lipase activity. Growth hormone is lipolytic, and the effects of recombinant human growth hormone (rhGH) on the hypercholesterolemia of CRI are unsettled. To test this question, we gave rhGH for 14 days at a dosage of 3 units/day intraperitoneally to two-stage, 5/6 nephrectomized, male Sprague-Dawley rats (n = 18) compared to sex- and age-matched control (n = 27) and CRI (n = 40) rats. At the end of the study, CRI rats and those treated with rhGH had a similar degree of renal impairment, as assessed by serum concentrations (mean +/- SEM) of urea nitrogen (49 +/- 3 vs. 54 +/- 4 mg/dl), creatinine (0.9 +/- 0.0 vs. 1.0 +/- 0.1 mg/dl) and cumulative food intake (311 +/- 8 vs. 290 +/- 12 g). Serum urea nitrogen (16 +/- 4 mg/dl) and creatinine (0.4 +/- 0.1 mg/dl) concentrations as well as food intake (412 +/- 9 g) of control rats were significantly (p < 0.0001) different. Serum cholesterol concentration of CRI rats treated with rhGH (87 +/- 3 mg/dl) was not higher than those of CRI rats (81 +/- 2 mg/dl, p < 0.1338) but was significantly higher than in control rats (55 +/- 3 mg/dl, p < 0.0001). CRI rats treated with rhGH showed a similar serum albumin concentration and lower serum glucose than CRI rats (0.9 +/- 0.1 vs. 0.9 +/- 0.0 g/dl and 144 +/- 4 vs. 163 +/- 3 mg/dl, p < 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)


Impaired diluting segment chloride reabsorption in patients with cystic fibrosis.

Clearance studies were performed in 12 patients with cystic fibrosis (CF) during maximum water diuresis. The fractional diluting segment reabsorption was calculated from chloride clearance (CCl) and maximum free water clearance (CH2O) as 'CH2O/CCl+CH2O'. Fractional delivery to the diluting segment and from the proximal tubules was estimated by maximum urine flow during water diuresis (Vmax/GFR) and lithium clearance (CLi/GFR), respectively. The data were compared with results obtained in young healthy subjects. To estimate free water generation beyond the medullary diluting segment, CH2O/GFR was also measured during acute furosemide administration in 5 patients. Values for the term CH2O/CCl+CH2O were significantly lower in patients with CF (79.1 +/- 5.1%) as compared with healthy controls (89.4 +/- 4.4%). This was also true for the terms Vmax/GFR (CF: 9.5 +/- 1.6%; control subjects: 13.8 +/- 2.5%) and CLi/GFR (CF: 22.2 +/- 4.1%; control subjects: 30.7 +/- 5.1%). Compared to data derived from the literature, the solute-free water generation during administration of furosemide (CH2O/GFR) was significantly reduced in patients with CF (6.8 +/- 1.5%) as compared with healthy subjects (13.3 +/- 6.1%). We conclude that reabsorption in the diluting segment is impaired in patients with CF, perhaps at a site beyond the thick ascending limb of Henle's loop. Enhanced proximal reabsorption, suggested by reductions in Vmax/GFR and CLi/GFR, appears to be a compensatory phenomenon to maintain sodium and chloride balance.


Association of microalbuminuria with slow acetylator phenotype in type 1 diabetes mellitus.

The genetically determined acetylator phenotype in diabetic children with and without increased urinary albumin excretion was investigated. Acetylator phenotype was determined according to Evans, and 24-hour albumin excretion rate (AER) was measured by immunoturbidometry in 86 children and adolescents with type 1 (insulin-dependent) diabetes mellitus and in 100 age-matched healthy controls. In diabetics, the fast acetylator phenotype was found in 36 (41.9%) patients and the slow one in 50 (58.1%); the control group had 52 (52%) fast and 48 (48%) slow acetylators. There were no significant differences in acetylator phenotypes between diabetic patients and control subjects (chi 2 = 1.0, NS). Among patients with normal albumin excretion (n = 70, mean age: 12.9 +/- 3.5 years, mean diabetes duration: 5.3 +/- 3.8 years, AER < 20 micrograms/min), 35 (50%) fast acetylators and 35 (50%) slow acetylators were found. In patients with elevated albumin excretion (n = 16, mean age: 14.0 +/- 3.2 years, mean diabetes duration: 4.9 +/- 3.0 years, AER > 20 micrograms/min), 1 (6.3%) patient was a fast acetylator and 15 (93.7%) were slow acetylators. A significant difference has been found between the two groups in the rate of fast/slow acetylators (chi 2 = 8.79, p < 0.01). The strong correlation between the slow acetylator phenotype and microalbuminuria in diabetics suggests that: (a) genetic factors may play a role in the development of diabetic nephropathy; (b) the acetylator status could be a useful tool to detect patients 'at risk' of nephropathy.


Evoked potentials before and after anemia correction with recombinant human erythropoietin in end-stage renal disease.

Subclinical involvement of the nervous system in uremic adults has been detected by modern neurophysiological techniques. Chronic anemia is one of the possible factors responsible for neural dysfunction in uremia. We evaluated neurophysiological (brainstem auditory and somatosensory evoked potential) abnormalities and their possible modification following anemia correction with recombinant human erythropoietin in 14 children with end-stage renal disease maintained by hemodialysis. Only peripheral and 8th cranial nerve electrophysiological data are significantly abnormal in our patients, and they are not acutely modified by anemia correction. These data confirm the importance of electrophysiological testing of uremic children for detecting nervous system involvement at an early stage and for monitoring the efficacy of its management.


Delayed pubertal growth spurt in glomerulopathic boys receiving alternate-day prednisone.

Statural growth was studied in 16 glomerulopathic children, 12 boys and 4 girls, treated with alternate-day prednisone for more than 2 years. Five patients were older than 12 years when started on therapy. Growth velocity was significantly slower in boys who received therapy during peripubertal age than in younger boys taking similar amounts of prednisone. In boys, pubertal growth spurt was delayed and shorter than normal, but growth continued with a velocity greater than normal for some years and kept on after 19 years of age. The ultimate height of the patients followed up after 19 years of age ranged within the normal limits of genetic height potential.


Residential summer camp for children with end-stage renal disease.

Residential summer camps exist for children with all varieties of chronic illness with the goal of improving their quality of life. This paper describes the development and implementation of a summer camp for children 9-18 years old who receive long-term peritoneal dialysis or who have received a kidney transplant. Thirty-five to forty children regularly participate in activities such as water olympics, survival hikes and campouts while continuing to receive their medical needs from trained personnel. A study to evaluate the impact of a summer camp revealed less patient hopelessness and improved self-esteem following the 1-week camping experience. Attendance at camp provided the medical staff with a unique perspective of childhood illness, while the period of respite for the parents was uniformly welcomed and may contribute to the prevention of parent burnout. It is hoped that the success of this camp and others like it will lead to the development of similar experiences for other children with chronic disease.


Efficacy of CAPD as the primary treatment for end-stage renal failure in children.

In the 6 years before this paper, all children presenting with end-stage renal failure (ESRF) in Northern Ireland were electively commenced on continuous ambulatory peritoneal dialysis (CAPD). In that period, 22 patients were treated, 16 received renal transplants and 6 remained on CAPD. Active treatment was withdrawn in 1 child due to obliterative peritonitis. There were no deaths. One incident of peritonitis was documented per 11.6 patient treatment months. Recurring peritonitis lead to catheter replacement in 3 patients. Other problems encountered were catheter exit site infections, hernia formation and in one instance, hydrothorax. Peritonitis was more common in children under 2 years of age as was undernutrition, growth retardation and developmental delay. Home CAPD can be recommended as a viable first option treatment for childhood ESRF where suitable haemodialysis facilities are not easily accessible.


Primary hyperoxaluria type 1: clinical manifestations in infancy and prenatal diagnosis.

A 9-month-old Pakistani boy of consanguineous parents presented with uraemia preceded by pyuria from 5 weeks of age. He had no history of renal calculi or macroscopic haematuria. Renal biopsy revealed severe calcium oxalate deposition in the tubuli and fibrosis of the interstitial tissue. Liver biopsy demonstrated complete absence of alanine: glyoxylate aminotransferase catalytic activity and immunoreactive protein compatible with a diagnosis of primary hyperoxaluria type 1. He died at the age of 11 months, just before liver transplantation was made possible. Fetal liver biopsy in the mother's subsequent pregnancy showed normal enzymatic activity. Early detection and early replacement of the missing enzyme by liver transplantation are considered to be crucial for the survival of severely affected infants with the acute neonatal form of primary hyperoxaluria type 1. Persistent pyuria could be an early sign of renal damage secondary to accumulation of oxalate crystals.


Sudden death in cystinosis.

We report on an 8-year-old boy with infantile cystinosis. The patient died in hospital for unknown reason after cardiopulmonary arrest although resuscitation was started immediately. As opposed to other cases [1-3], neither laboratory investigations nor autopsy in out patient gave an explanation for the sudden death.


Conformational analysis of beta-D-fructofuranosyl-(2-->6)-beta-D-glucopyranoside by molecular mechanics (MM2) calculations.

Conformational energies for models of the disaccharide beta-D-fructofuranosyl-(2-->6)-beta-D-glucopyranoside were computed by molecular mechanics using MM2(87). An initial investigation of staggered forms examined the linkage bonds characterized by the torsion angles phi, psi, and omega, and subsequently the fructose hydroxymethyl side groups, characterized by the torsion angles chi-1 and chi-6. Then, in our major search of conformational space, the torsion angles of two linkage bonds, phi and omega, were driven through 360 degrees in 20 degree increments at all staggered side group combinations. From these results, the low-energy forms were minimized without the driver restrictions to generate the global minimum structure found and herein reported. This conformer was then used to map the conformational space of phi and omega by driving only those torsion angles through 360 degrees. Both the 4(3)T (northern) conformer (Cremer-Pople puckering phase angle of phi 2 = 265 degrees) and the 3(4)T, (southern) conformer (phi 2 = 80 degrees) of the fructofuranose ring were tested for comparison, and both were shown to be significant contributors of populated forms. As these two conformers had different minima for a number of important torsion angles, experimental studies may reveal different properties than those expected solely from the preferred northern conformer.


Sulfated sialyl-oligosaccharides derived from tracheobronchial mucous glycoproteins of a patient suffering from cystic fibrosis.

Thirteen novel oligosaccharides, each possessing both a sulfate ester and a sialic acid residue, were isolated from tracheobronchial mucous glycoproteins from a patient with cystic fibrosis via cleavage by alkaline borohydride treatment, and by employing immobilized Limulus polyphemus lectin affinity chromatography, SynChroprep AX300 anion-exchange chromatography, Bio-Gel P-2 size-exclusion chromatography, and Hypersil 120A APS-2 high-performance liquid chromatography (HPLC). Proposed structures for the resulting purified sulfated sialyl-oligosaccharides were based on carbohydrate/permethylation analyses, periodate oxidation, complete sequential exoglycosidase digestion, analysis of desulfated products and, analysis by positive-ion fast-atom-bombardment mass spectrometry (FABMS). Sulfate esters on these sialyl-oligosaccharides resided on C-6 of a terminal or an internal D-galactose or 2-acetamido-2-deoxy-D-glucose residue or C-4 of a terminal D-galactose residue. The sialic acid residues were found to be either bound (2-->6)-alpha to 2-acetamido-2-deoxy-D-galactitol or (2-->3)-alpha or (2-->6)-alpha to a D-galactose residue occupying a nonreducing terminus. For this group of oligosaccharides, ranging in size from tri- to hepta-saccharides, it was also observed that a sialic acid residue and a sulfate ester did not residue on the same oligosaccharide branch when more than one branch existed. On linear unbranched sulfated sialyl-oligosaccharides, the sialic acid residue was bound to a D-galactose residue occupying a nonreducing terminus with the sulfate ester residing on an internal D-galactose or a 2-acetamido-2-deoxy-D-glucose residue. These results demonstrate that it is possible for sialic acid and a sulfate ester to exist on the same oligosaccharide and that this oligosaccharide can be as small as a trisaccharide.


The capsular antigen of Escherichia coli serotype O8:K102:H-.

The structure of the capsular antigen of E. coli O8:K102:H- was investigated by methylation analysis, beta-elimination of the methylated polysaccharide, lithium-ethylenediamine mediated degradation, and by 1D and 2D 1H and 13C NMR spectroscopy of the lithium-degraded and native polysaccharides. The capsular antigen was shown to have the following branched pentasaccharide repeating unit: [formula: see text]


The structure of the capsular polysaccharide of Escherichia coli O9:K35:H-.

The annexed structure of the capsular polysaccharide of Escherichia coli O9:K35:H- (A104a) has been determined by glycose and methylation analysis, base-catalysed degradation, and 1D and 2D NMR spectroscopy on the polysaccharide and the products obtained by bacteriophage-mediated depolymerisation. [formula: see text]


The structure of the fructan sinistrin from Urginea maritima.

The structure of sinistrin from red squill (Urginea maritima) was determined by methylation analysis and 13C NMR spectroscopy, using the fructans from Pucinella peisonis and quack-grass (Agropyron repens) as reference substances. Application of the reductive cleavage method showed that, of the beta-D-fructofuranosyl residues in sinistrin, 33% were 1-linked, 19% were 6-linked, 25% were terminal, and 19% were 1,6-linked. The average dp was 31 and, of the 3.24% of alpha-D-glucopyranosyl residues, 0.54% were terminal and 2.70% were 6-substituted. The fructan of quack grass was also highly branched with a (2-->6)-linked backbone, terminal alpha-D-glucopyranosyl residues, and a dp of approximately 45. The fructan from Pucinella peisonis was slightly branched, with a dp of approximately 10 and a (2-->6)-linked backbone.


Fructo-oligosaccharides from the stems of Triticum aestivum.

Fructo-oligosaccharides, extracted with hot water from wheat stems before flowering, were fractionated by gel-permeation chromatography on Biogel P2. The tri-/tetra- and penta-saccharide fractions were purified by HPLC and analysed by the reductive cleavage method. The trisaccharides 1-kestose and 6-kestose, the branched tetrasaccharide bifurcose, and the (2-->6)-linked pentasaccharide were identified. The fractions of higher molecular weight were also investigated and confirmed the branched-levan structure of this fructan.


Development of a water-soluble, sulfated (1-->3)-beta-D-glucan biological response modifier derived from Saccharomyces cerevisiae.

This report describes a method for the solubilization of micro-particulate (1-->3)-beta-D-glucan. Insoluble glucan is dissolved in methyl sulfoxide and urea (8 M) and partially sulfated at 100 degrees. The resulting water-soluble product is called glucan sulfate. The conversion rate is 98%, and the preparation is endotoxin free as determined by the Limulus lysate procedure. Glucan sulfate is composed of 34.06% C, 6.15% H, 50.30% O, 5.69% S and 3.23% N, and has a repeating unit empirical formula of (C6H10O5)8.3 SO3NH4+.4 H2O, suggesting that, on the average, a sulfate group is substituted on every third glucose subunit along the polymer. Molecular weight averages, polydispersity, and intrinsic viscosity were determined by aqueous high-performance size-exclusion chromatography (HPSEC). Two polymer peaks were resolved. Peak 1 (Mw = 1.25 x 10(6) g/mol) represents < 1% of the total polymer mass. Peak 2 (Mw = 1.45 x 10(4) g/mol) comprises > 99% of polymers. 13C NMR spectroscopy confirmed the beta-(1-->3) interchain linkage. In solution, glucan sulfate polymers self-associate in a triple helix. Glucan sulfate stimulates murine bone marrow proliferation following intravenous administration. The ability to prepare a immunologically active, water-soluble (1-->3)-beta-D-glucan preparation will greatly enhance the clinical utility of this class of compounds.


Mechanism of action of D-galacturonan digalacturonohydrolase of Selenomonas ruminantium on oligogalactosiduronic acids.

The mechanism of action of the specific D-galacturonan digalacturonohydrolase [poly-(1----4)-alpha-D-galactosiduronate digalacturonohydrolase, EC 3.2.1.82] of Selenomonas ruminantium was investigated by using reducing-end [1-3H]-labeled oligogalactosiduronates having degree of polymerization 3-5 as the substrates. The reaction products, incorporation and distribution of radioactivity in products, and the frequency of oligogalactosiduronate bond-cleavage were quantitatively estimated as functions of the substrate concentration. An alternative cleavage of tri(D-galactosiduronate) occurred during the enzyme reaction, indicating the participation of some bimolecular mechanism in addition to unimolecular hydrolysis in the action of the enzyme. Unimolecular hydrolysis takes place at low initial concentration of the substrate. The shifted termolecular enzyme-substrate complex formation and the subsequent galactosyluronic transfer is the predominant mechanism in degradation of tri(D-galactosiduronate) at high concentration. Tetra(D-galactosiduronate) and penta(D-galactosiduronate) are degraded by unimolecular hydrolysis at low, as well as high concentration of the substrate.


A 1H NMR database computer program for the analysis of the primary structure of complex carbohydrates.

A 1H NMR database computer program has been developed to determine the primary structure of complex carbohydrates. The database contains carbohydrate structures, their corresponding 1H NMR data, and literature references. From an input list of chemical shift values, the program generates an output list of partially or completely matching carbohydrate structures. In order to facilitate the recognition of the matching part of the selected carbohydrate structures, these structures are displayed with the matching structural elements highlighted. This new 1H NMR database, together with the search program described, now provides a fast access to the published 1H NMR data of complex carbohydrates and furnishes easy links to carbohydrate structures. The performance of the program is demonstrated by the analysis of five carbohydrate fractions prepared from a pool of horse serum glycoproteins.


A 1H and 13C NMR study of oligosaccharides from human milk. Application of the computer program CASPER.

Several oligosaccharides from human milk, containing vicinally branched residues, have been analysed with respect to induced NMR chemical shift changes that originate from the branching. Two types of branching were investigated: (i) linear oligosaccharides with a 2-linked residue, which thus becomes vicinally 1,2-disubstituted, and (ii) oligosaccharides with either 2,3- or 3,4-branching. It could be concluded that, in 13C NMR spectra of the first type, for which only moderately sized induced changes (< 2 ppm) had been observed previously, large (> 5 ppm) changes are also present. For 2,3- and 3,4-branching, changes similar to those observed earlier were found. In 1H NMR spectra, significant induced shifts for signals from anomeric, aglyconic, and H-5 protons were observed. For most trisaccharides, a unique set of values for the chemical shift differences was found, thus making it suitable to use them for characterisation of substitution patterns in the analysis with the computer program CASPER.


Fmoc-protected, glycosylated asparagines potentially useful as reagents in the solid-phase synthesis of N-glycopeptides.

1-Amino 1-deoxy derivatives of unprotected O-beta-D-galactopyranosyl-(1-->3)-2-acetamido-2-deoxy-beta-D-glucopyrano se, 2-acetamido-2-deoxy-D-galactose, D-galactose, lactose, D-fucose, D-mannose, and 2-deoxy-D-arabino-hexose were prepared and acylated with N-fluorenylmethoxycarbonylaspartic acid alpha-tert-butyl ester. The anomeric configuration of the N-glycosyl bond (including that of the mannose derivative) in each of the purified compounds was shown to be beta. The probable stability of the N-glycosyl and glycosidic bonds during the conditions of solid-phase peptide synthesis was investigated by treatment of the glycosylated asparagine derivatives with different concentrations of trifluoroacetic acid. Based on their stability, we found that Fmoc-Asn(sugar)-OH derivatives are excellent candidates for automated synthesis of biologically active glycopeptides.


Stereoselective total synthesis of the glycosyl phosphatidylinositol (GPI) anchor of Trypanosoma brucei.

The total synthesis of O-(O-[6-O-(2-aminoethylphosphono)-alpha-D-mannopyranosyl]-(1-->2)- O-alpha- D-mannopyranosyl-(1-->6)-O-[O-alpha-D-galactopyranosyl-(1-->6)-alpha-D- galactopyranosyl-(1-->3)]-O-alpha-D-mannopyranosyl-(1-->4)-2-amino-2-deo xy- alpha-D-glucopyranosyl)-(1-->6)-[1-O-(1,2-dimyristoyl-sn-glycero-3-phosp hono) - 1D-myo-inositol], the GPI anchor of Trypanosoma brucei was achieved for the first time. The core structure of the GPI molecule, the glycoheptaosyl part, was constructed in a highly stereocontrolled manner from O-[O-(2,4-di-O-benzyl-alpha-D-mannopyranosyl-(1-->4)-2-azido-3,6-di-O-be nzyl- 2-deoxy-D-glucopyranosyl]-(1-->6)-2,3,4,5-tetra-O-benzyl-1-O-(4- methoxybenzyl)-D-myo-inositol, O-(2,3,4,6-tetra-O-benzyl-alpha-D-galactopyranosyl)-(1-->6)-2,3,4-tri-O- benzyl-D-galactopyranosyl fluoride, 2-O-acetyl-3,4,6-tri-O-benzyl-alpha-D-mannopyranosyl chloride, and 6-O-acetyl-2,3,4-tri-O-benzyl-alpha-D-mannopyranosyl fluoride. The introduction of two phosphodiester functions was efficiently achieved using the H-phosphonate method.


Tobacco smoking, alcohol consumption, and laryngeal cancer in Madrid.

The associations between cigarette smoking and alcohol consumption and laryngeal cancer were examined in a case-control study carried out between 1982 and 1985 in Madrid. The analysis was based on 50 histologically confirmed male cases and 103 age- and sex-matched controls (45 hospitalized and 58 from the general population). A dose-response effect was observed for cigarette smoking, with an odds ratio (OR) of 4.33 (95% confidence interval of 1.22 to 15.41) for smokers of 30 or more cigarettes per day, when compared with those smoking less than 10 cigarettes per day. The risk clearly rose in line with the length of the smoking habit. In addition, for smokers of black tobacco, the risk was more than double that for smokers of blond tobacco, irrespective of the depth of inhalation. ORs for alcohol consumption rose significantly in accordance with the average grams intake per week and the overall lifetime consumption, but not with years of drinking. The time trends of risks for duration of alcohol consumption suggest the existence of phenomena related to individual susceptibility. A dose response effect was observed in supraglottal and glottal tumors. The effect of the joint exposure to both tobacco and alcohol fit to a multiplicative model.


Calculated risk of breast cancer following mantle irradiation determined by measured dose.

Tumor registry data indicate a two- to fourfold increased incidence of breast cancer following mantle irradiation, but cumulative risk is unknown. Radiation exposure to the breasts underlying the mantle block ranges from 4 to 40 Gy and is dependent on relative positions of the breasts and mantle block. Unshielded outer breast quadrants near axillary nodal regions receive 36 to 40 Gy, while central breast quadrants under the lung blocks receive approximately 4 Gy as determined by dose volume histogram analysis. Relative dose risk analysis for breast cancer following mantle irradiation was performed and indicated an overall excess risk of 1.5 for the upper outer quadrant (total dose 40 Gy), 1.3 for the upper and lower inner, and central quadrants (total dose 15 to 20 Gy), and 1.2 for the lower outer quadrant (total dose 4 Gy). Linear and cell-kill carcinogenesis models demonstrated similar relative risk assessments in the low-dose regions, defined as < 15 Gy. Predicted risk for breast cancer in the high-dose regions (> or = 15 Gy) varied considerably according to the model evaluated. The linear model predicted a three to ten times greater risk above baseline breast cancer incidence for the high-dose regions. In contrast, the cell-kill model predicted no excess cases of breast cancer, assuming cell death at these higher dose levels. The greatest relative predicted risk is observed in women < 20 years of age at the time of irradiation; however, women older than 20 years continue to have a 50% higher than baseline risk for subsequent breast cancer development. All women treated for Hodgkin's lymphoma should undergo dose volume histogram evaluation. Prospective clinical and mammographic evaluations should be performed in all female patients following mantle irradiation to better define the risk for secondary breast carcinogenesis.


Risk of leukemia among participants of gastric mass screening survey in Japan: a population-based, case-control study.

To assess the risks of leukemia for individuals participating in a gastric mass screening program using barium photofluorographic examination in Japan, a population-based, case-control study was carried out. The histories of participation in the last 10 years of 79 leukemia cases diagnosed between 1978 and 1985 and twofold controls matched for sex, age (+/- 5 years), and residence were investigated using the records of the gastric mass screening program. The odds ratio of any type of leukemia for those participating at least once in the last 10 years was 1.09 (95% confidence interval, 0.61-1.96) and that of myeloid type was 0.88 (95% confidence interval, 0.38-2.27). Even when the histories of the preceding 5 years were ignored, no significant excess risks were found among patients with any type and myeloid type of leukemia. These results suggest that there is insufficient evidence to support the positive association between the incidence of leukemia and participation in gastric mass screening. The limitations and implications of this analysis are discussed.


Clinical characteristics of patients with epidermoid carcinoma of the upper aerodigestive tract who develop second malignant tumors.

A total of 1294 patients with primary head and neck (H&N) cancer of a single site was diagnosed during the years 1970 to 1979 at the Memorial Sloan-Kettering Cancer Center, 163 subsequently developed a second malignant tumor (SMT). In 50% of the cases, the second tumor was in the H&N, and in 30% in the lung and esophagus; 41 patients developed a third tumor. Again, in 50% of these cases, the tumor was in the H&N and in 17% in the lung and esophagus. The stage of disease of all the second primary tumors was more advanced at diagnosis, mainly as a result of the large number of patients with lung and esophagus cancer. Patients who had a second tumor in the H&N were diagnosed in an earlier stage of disease than patients with a single H&N tumor. The survival of patients with localized second H&N cancer was worse than for those with a primary, localized single tumor.


Cancer-associated SCM-recognition, immunedefense suppression, and serine protease protection peptide. Part I. Isolation, amino acid sequence, homology, and origin.

The isolation of a cancer-associated, SCM-recognition, immunedefense-suppressing, and serine protease-protecting (CRISPP) peptide from the blood plasma of cancer patients is described. The amino acid sequences were determined on preparations from 12 different cancers. The peptide is composed in 9 cancers of 29 and in 3 cancers of 35 amino acid residues with molecular weights of 3410 and 4007 Da, respectively. A consensus, synthetic 29 amino acid CRISPP peptide (CRISPPs) has the same cancer SCM-recognition (CR) activity and SCM-response modifying effects as the natural peptide. The "cancer SCM-recognition epitope" of the CRISPP peptide was determined. Anti-CRISPPs antibodies were raised and used in immunoassays to confirm the presence of the CRISPP peptides in cancer blood plasmas, in supernatants of cancer cell growth media and in cultured human cancer cells. The amino terminal end sequences of peptides isolated from growth media of cultured breast and colon cancer cells corresponded to amino acid sequences of CRISPP peptides isolated from cancer blood plasmas of subjects with the respective cancers. The CRISPP peptides are between 83 to 100% homologous to the alpha 1-protease inhibitor amino acid sequence located at the carboxy terminal end between residues 358 and 393. The genetic origin of the CRISPP peptides and their selective advantage to cancer cell survival are discussed.


Strategic approaches to cancer control research in NCI-funded research bases.

Although the Community Clinical Oncology Program (CCOP) of the National Cancer Institute (NCI) has always encouraged research on cancer prevention and control issues, the clinical cooperative groups and cancer centers that serve as CCOP "research bases" were not required to conduct cancer control research until June 1987. This article draws upon case studies of six cooperative groups and two cancer centers to examine how CCOP research bases have defined cancer control research to be compatible with their missions, restructured roles and relationships to facilitate protocol development and implementation, and routine cancer control research to make prevention and control studies an accepted component of their scientific programs. Three "deliberate" or "emergent" strategies are found to be associated with the earlier resolution of problems and more rapid assimilation of cancer control studies into research base agendas. These strategies include (1) proactive planning, such as the early formation of a Cancer Control Research Committee (CCRC) and the identification of "idea champions" to promote cancer prevention and control research; (2) a staged approach to defining and introducing cancer control research; and (3) the creation of multiple "bridging" mechanisms to link cancer prevention and control studies with ongoing research activities. Research bases were most successful in building investigator commitment to cancer control research when they framed research questions within an accepted scientific paradigm, sought out and supported investigators with an interest in cancer prevention and control questions, created CCRCs with broadly representative memberships, and widely publicized ideas for prevention and control studies.


Barriers to cervical cytology screening in native women in British Columbia.

The British Columbia (BC) Cervical Cytology Screening Programme (CCSP), implemented since 1955, has resulted in a 75% decrease in both the mortality from and incidence of invasive squamous cervical carcinoma. However, despite this effect, the Native Indian population still present an overall mortality rate four times higher than that of the non-Native population. A demonstration project was initiated in four Native Indian Band communities of BC to determine the reasons underlying these findings. The participation patterns to the CCSP were investigated and revealed that the overall percentage of participation to the CCSP among Native Indian women was 30% lower than that of the non-Native population. Reasons for the under-participation to the CCSP were explored. A total of 36 women, 9 in each of the 4 communities, including current users, ex-users and never users, were interviewed. Reasons for not participating in the CCSP were due mainly to (1) the lack of knowledge about the Pap test and about its importance; (2) feelings of embarrassment and shamefulness; (3) lack of continuity of care due to the high turnover of physicians in the Native communities. Based on the study findings, a pilot CCSP clinic will be implemented in each community. In addition to taking cervical smears, this pilot project will include education sessions, notification about the results of the test, and recall for annual check up.


Clinical and virological aspects of HIV2 infection in rhesus monkeys.

To establish an animal model of AIDS, two different "wild" or "adapted" HIV2 Rod and Eho strains were cultivated on monkey cells from different species (baboons, cynomolgus, Rhesus monkeys). Five different available strains were then injected both by intravenous (i.v.) and intracerebral (i.c.) route into ten Rhesus monkeys. Seven animals seroconverted between days 13 and 230. Reverse transcriptase activity in the lymphocyte culture supernatants was detectable in six of the seven animals that seroconverted, and in one animal that remained seronegative. Lymphopenia and a decrease in the CD4+ cell counts were observed in eight animals. One animal, inoculated with HIV2-Rod "wild type," developed a severe cachexia, with dyspnea, and associated neurological symptoms 150 days after inoculation. This animal was sacrificed on day 220. Pathological examination showed typical lesions of actinomycetes infection in the lungs and in the meninges. Another monkey had significant weight loss associated with lymphadenopathies and pancytopenia. These results suggest that in vivo replication of HIV2 in Rhesus monkeys may induce clinical symptoms of immune deficiency. This method is reproducible and may provide a good model for AIDS.


The modeling of breast cancer deaths in Vermont.

Methodologic issues in modeling breast cancer deaths were investigated through modeling the number of breast cancer deaths in Vermont for the period 1975 to 1988. Age-specific incidence rates, case fatality rates, and secular trends of these rates were necessary to represent the observed trend in breast cancer deaths over this time period. Additional information, such as mortality from other causes, stage distribution, or screening history, was not necessary. Incidence and survival information were obtained from a statewide population-based breast cancer registry in Vermont and the Surveillance, Epidemiology and End Results (SEER) program. SEER incidence rates overestimated the actual number of deaths for long-term survival rates. Case fatality rates and secular trends in incidence and survival reported by SEER were appropriate. These results can be applied to the planning of community intervention studies by providing the essential expected numbers of deaths in control communities.


[Hemodialysis in diabetics].

In the submitted review the author deals with the problem of haemodialysis in diabetics. He summarizes some basic information on the state of haemodialysis treatment of diabetics in advanced countries with regard to the situation in Czechoslovakia, the survival of patients and causes of mortality, complications, the standard of rehabilitation and quality of life. The author gives a rough outline of the strategy of care of diabetics in the pre-dialysis stage of diabetic nephropathy and in a chronic dialysis programme. The author emphasizes that haemodialysis means a significant prolongation of life in patients with advanced diabetic nephropathy. In an integrated dialysis transplantation programme it is possible to achieve in these patients prolonged satisfactory quality of life after successful transplantation.


[The Prague Diabetes Registry. Computer utilization: basic information on a group of diabetics and patients with impaired glucose tolerance].

In 1989 the authors founded in Prague a register of all diabetics and patients with IGT. The basic data on each patient are in a computer and are systematically supplemented. The mean age of the investigated diabetics is 68.8 years in women and 64.7 years in men. The prevalence of DM and IGT in Prague women is higher (4.8%) than in men (4.3%). Practically half the Prague group suffers from diabetes for less than 5 years and only 4.6% of the diabetics are on the records for more than 20 years. DM type 1 was manifested in 54.7% of the patients at an age under 29 years and the majority of type 2 diabetics in Prague are in the age bracket of 60 to 69 years. The incidence of DM in 1988 to 1990 remains equal and amounts to cca 5%.


[The effect of fish oil on the secretion and effect of insulin in patients with type II diabetes].

The metabolic effect of 3-week dietary supplementation with a fish oil concentrate was examined in not markedly obese, not hypertriglyceridemic men with non-insulin-dependent diabetes mellitus (NIDDM) treated with hypoglycemic agents. Ten patients were given 15 ml/d of fish oil (Martens Oil, Norway) equivalent to 3.1 g of n-3 fatty acid (FA) per day, and compared to 10 diabetics treated with placebo (15 ml/d saline). While fish oil leads to expected increase in the ratio of n-3 to n-6 FA intake, it does not alter fasting and mixed meal stimulated blood glucose, plasma insulin and C-peptide concentrations. There were no changes in insulin action estimated by the metabolic clearance rates of glucose at plasma insulin levels of about 100 microU/ml and 1400 microU/ml during hyperinsulinemic isoglycemic clamp, and no changes were seen in insulin binding to erythrocytes. Even though our short-term study does not warrant authoritative conclusions, no adverse effects of low-dose fish oil on glucose homeostasis have been found in not markedly obese NIDDM patients treated with oral hypoglycemics.


[Diagnosis of conditions following myocardial infarct using complex analysis of electrical cardiac fields].

In 22 patients with ischaemic heart disease and conditions after infarction and angina pectoris a coronarographic examination was made as well as other auxiliary examinations incl. a complex analysis of the electrical cardiac field (KAESP) (23), using a Cardiac apparatus (manufacturer ZPA Cakovice). Using the KAESP method fibroses in the heart muscle were found in all 22 patients, while a classical electrocardiogram revealed them only in 12 patients (54.5%). Post-infarction fibrous changes on the inferior cardiac wall were detected by ECG in 10 patients, KAESP revealed this localization of changes in 17 patients. The difference was particularly marked as regards localization on the anterior cardiac wall, according to ECG it was in 2 patients, according to KAESP in 17 patients. The authors investigated also on isopotential repolarization maps focal changes caused by cardiac ischaemia associated with organic affection of the appropriate coronary artery as revealed by coronarography. Identical sites were proved in 18 patients by the two methods, i. e. in 81.8%. In the discussion the authors analyse the causes which influence the accuracy of assessment of the coronary artery in KAESP. In KAESP in addition to isopotential maps also other maps were used such as isointegral, iso-areal, asynchronic potential maxima and minima, isochronic maps, maps of negative isodivergencies, profile sections etc. (20).


Headache in ischemic cerebrovascular disease. Part II: Mechanisms and predictive value.

In part I, the prevalence and clinical characteristics of headache in ischemic cerebrovascular disease were reviewed. In this part, we describe the potential mechanisms of head pain and the value of the headache symptom as a predictor of stroke type, location and vascular territory.


Visual field loss after attacks of migraine with aura.

Visual fields were mapped with kinetic are perimetry in 23 migraine with aura subjects and, for comparison, in 20 migraine without aura subjects and in 21 non-headache controls. Central vision on the Amsler eye chart and visual perception threshold on a computer task were also investigated. Measures were obtained at least seven days after an episode of migraine. In addition, 10 of the migraine with aura subjects and 10 migraine without aura subjects were studied the day after an attack. The day after migraine with aura, visual sensitivity in the periphery of the visual fields was depressed, central vision was blurred, and visual perception threshold was elevated. These visual disturbances had resolved 7 to 10 days later. With the exception of a minor increase in visual perception threshold, vision was normal after attacks of migraine without aura. Residual effects of the migraine aura could mediate the subclinical visual disturbances which persist for at least one day after attacks of migraine with aura.


Benign coital headache.

We studied the natural history of patients with a diagnosis of benign coital headache who presented to a private neurological clinic between the years 1978 and 1991. Thirty-two patients (24M, 8F) were invited to participate and 26 patients (83%) responded. The period of follow-up ranged from six months to 14 years (median 6 years). Thirteen patients (50%) had recurrent attacks of coital headache epochs separated by intervals of up to 10 years. Eleven of these patients suffered a concomitant primary headache whereas this was present in only one of those patients without recurrent attacks of coital headache (p < 0.001). In all but one patient, who had a transient blurred vision, the headache was not accompanied by nausea, vomiting, visual disturbances, sensory/motor disturbances, or unconsciousness. We concluded that benign coital headache can be clearly distinguished from headaches due to cerebral aneurysm or arteriovenous malformation rupture. The presence of a concomitant primary headache syndrome is a risk-factor for recurrence of coital headache.


Non-migraine headache across the menstrual cycle in women with and without premenstrual syndrome.

Fluctuation of estrogen levels across the menstrual cycle influences migraine headache. In this study, 53 women documented prospectively the incidence and severity of headache daily for an average of three menstrual cycles. Seven of the women met the criteria established by the International Headache Society for migraine with or without aura, while the remaining 46 women failed to do so. Chi-square analysis revealed that, overall, the incidence of non-migraine headache was dependent on day of the cycle (chi 2 [1,66] = 247.7, p < 0.001), with more headaches occurring during the perimenstrual phase. The 46 women without migraine were further classified according to NIMH criteria into PMS (n = 26) and non-PMS groups (n = 20). An association between headache and menstrual cycle phase was noted for both groups (p < 0.001), although the incidence of severe headache was greater for the PMS women, during both the perimenstrual and intermenstrual phases. Both groups experienced an increase in severe headaches during the perimenstrual phase. The PMS women peaked on the day prior to menstruation, while the non-PMS women peaked on the first day of menstruation. There did not appear to be an overall difference in the reporting of mild headache across the cycle between women with or without PMS.(ABSTRACT TRUNCATED AT 250 WORDS)


Cardiovascular reflexes in patients with vascular headache.

We have investigated the autonomic function of 75 patients with migraine by examining cardiovascular reflex function. The results were compared with those of 78 healthy volunteers. Measurements were made between attacks. Patients with migraine showed a smaller heart-rate response to deep breathing but a greater heart-rate response and higher blood pressure to standing when compared to controls. Migraine patients had a higher percentage of established sympathetic lesions (51% vs 17%) and severe (25% vs 5%) or atypical (24% vs 11.5%) global autonomic dysfunction. No significant differences were found among patients with migraine with aura, migraine without aura, and migraine with prolonged aura. Our findings indicate that patients with migraine have sympathetic hypofunction.


Evaluation of chronic daily headache--comparison to criteria for chronic tension-type headache.

The purpose of this study was to evaluate the adequacy of the International Headache Society (IHS) criteria for chronic tension-type headache and, if appropriate, suggest modifications of the IHS classification. We evaluated 100 consecutive patients with chronic daily headache. Approximately two-thirds of our patients fulfilled the criteria for chronic tension-type headache. Most of the patients who failed to meet the criteria did so because they had more than one migrainous feature. Approximately 50% of patients took excessive amounts of analgesic medication. We conclude that the IHS criteria should be modified to include chronic daily headache evolving from migraine; subtypes with and without medication overuse should be distinguished.


Presentation of a new instrument: the diagnostic headache diary.

A new instrument, the Diagnostic Headache Diary, based on the operational diagnostic criteria of the International Headache Society (IHS), was tested in 61 migraine patients from a headache research clinic using the clinical diagnosis (IHS criteria) for comparison. All patients kept the diary for one to eight months. The clinical and diary diagnosis of migraine with and without aura was the same in, respectively, 72 and 87% of the patients. Nausea, photophobia and phonophobia tended to be more pronounced at the clinical interview. The diary identified 20 more cases of episodic tension-type headache and 15 fewer cases of chronic tension-type headache than the clinical interview. Two blinded observers always made the same IHS diagnoses when interpreting the diagnostic headache diary. A combination of a clinical interview and the diagnostic headache diary gives a qualitatively and quantitatively more precise diagnosis than a clinical interview alone.


Headache following carotid endarterectomy: a prospective study.

The occurrence of headache in the 28 days following surgery was studied in 50 consecutive patients (14F and 36M, mean aged 70 years) who underwent carotid endarterectomy for atheromatous carotid stenosis. Thirty-one patients (62%) reported headache. Headache occurred in the first five days after surgery in 87% of cases. Its characteristics and temporal profile were highly variable but it was mostly bilateral (74%), mild or moderate (78%), requiring no treatment (77%). No correlation was found between the occurrence of headache and degree of stenosis, intraoperative characteristics and past history of headache. In none of our patients was severe ipsilateral headache, cerebral hyperperfusion syndrome, or cluster-like hemicrania encountered and only five patients met the IHS criteria for post-endarterectomy headache. Post-endarterectomy headache is frequent when specifically looked for and is therefore not a single entity. The present IHS criteria are unsatisfactory and should be modified accordingly.


Transcranial Doppler ultrasonic features in chronic tension-type headache.

We studied vascular features in patients suffering from chronic headache of the tension-type by means of transcranial Doppler ultrasound (TCD). Blood flow velocities of the basal cerebral arteries and the submandibular extracranial part of the internal carotid artery were compared between 20 chronic tension-type headache sufferers and the same number of age- and sex-matched control subjects. There were no significant differences of ultrasonic features between the groups. Changes in blood flow velocities do not seem to be involved in the pathogenetic mechanisms of chronic tension-type headache.


Shortlasting, unilateral, neuralgiform headache attacks with conjunctival injection and tearing (SUNCT syndrome): V. Orbital phlebography.

We studied six patients with SUNCT, a unilateral headache syndrome with shortlasting attacks and ipsilateral autonomic phenomena with orbital phlebography and MRI. All but one orbital phlebogram showed abnormalities on the headache side (in one patient bilaterally), involving the superior ophthalmic vein or/and the cavernous sinus. No systematic changes were demonstrated on MRI. The phlebography findings were similar to those observed in the Tolosa-Hunt syndrome and in cluster headache.


High frequency mutagenesis by a DNA methyltransferase.

HpaII methylase (M. HpaII), an example of a DNA (cytosine-5)-methyltransferase, was found to induce directly a high frequency of C-->U transition mutations in double-stranded DNA. A mutant pSV2-neo plasmid, constructed with an inactivating T-->C transition mutation creating a CCGG site, was incubated with M. HpaII in the absence of S-adenosylmethionine (SAM). This caused an approximately 10(4)-fold increase in the rate of reversion when the mutant neo plasmid was transformed into bacteria lacking uracil-DNA glycosylase. The mutation frequency was very sensitive to SAM concentration and was reduced to background when the concentration of the methyl donor exceeded 300 nM. The data support current models for the formation of a covalent complex between the methyltransferase and cytosine. They also suggest that the occurrence of mutational hot spots at CpG sites may not always be due to spontaneous deamination of 5-methylcytosine, but might also be initiated by enzymatic deamination of cytosine and proceed through a C-->U-->T pathway.


The mammalian ultraviolet response is triggered by activation of Src tyrosine kinases.

Exposure of mammalian cells to DNA-damaging agents induces the ultraviolet (UV) response, involving transcription factor AP-1, composed of Jun and Fos proteins. We investigated the mechanism by which UV irradiation induces the c-jun gene. The earliest detectable step was activation of Src tyrosine kinases, followed by activation of Ha-Ras and Raf-1. The response to UV was blocked by tyrosine kinase inhibitors and dominant negative mutants of v-src, Ha-ras, and raf-1. This signaling cascade leads to increased phosphorylation of c-Jun on two serine residues that potentiate its activity. These results strongly suggest that the UV response is initiated at or near the plasma membrane rather than the nucleus. The response may be elicited by oxidative stress, because it is inhibited by elevation of intracellular glutathione. Using tyrosine kinase inhibitors, we demonstrate that the UV response has a protective function.


Activation of an inducible c-FosER fusion protein causes loss of epithelial polarity and triggers epithelial-fibroblastoid cell conversion.

As a novel approach to studying the modulation of the polarized epithelial phenotype, we have expressed c-Fos and c-Myc estrogen receptor fusion proteins (c-FosER and c-MycER) in mammary epithelial cells. The hybrid proteins could be activated by estrogen for defined time periods and after the cells had achieved their fully polarized organization. Activation of c-MycER deregulated proliferation but did not affect epithelial polarity. Short-term activation of c-FosER induced the reversible loss of morphological and functional cell polarity. In contrast, long-term stimulation of c-FosER caused the cells to depolarize irreversibly, to invade collagen gels, and to undergo epithelial-fibroblastoid cell conversion. Our data suggest that Fos proteins are important in modulating the epithelial phenotype both in normal tissue development and in invasive processes.


Lysosome recruitment and fusion are early events required for trypanosome invasion of mammalian cells.

Trypanosoma cruzi invades most nucleated cells by a mechanism distinct from classical phagocytosis. Although parasites enter at the lysosome-poor peripheral cell margins, lysosomal markers are immediately incorporated into the parasitophorous vacuole. No accumulation of polymerized actin was detected around recently internalized parasites, and disruption of microfilaments significantly facilitated invasion. Lysosomes were observed to aggregate at the sites of trypanosome attachment and to fuse with the vacuole at early stages of its formation. Experimentally induced, microtubule-dependent movement of lysosomes from the perinuclear area to the cell periphery enhanced entry. Conditions that deplete cells of peripheral lysosomes or interfere with lysosomal fusion capacity inhibited invasion. These observations reveal a novel mechanism for cell invasion:recruitment of lysosomes for fusion at the site of parasite internalization.


Endocytosis in yeast: evidence for the involvement of a small GTP-binding protein (Ypt7p).

From the budding yeast S. cerevisiae, we have cloned a gene, YPT7, that encodes a GTP-binding protein belonging to the Ypt family of ras-related proteins. The 208 amino acid protein shares identical effector domain and C-terminal sequences with the mammalian Rab7 protein. YPT7 gene disruption did not impair cellular growth at temperatures ranging from 17 degrees C to 37 degrees C. ypt7 null mutants are characterized by highly fragmented vacuoles and differential defects of vacuolar protein transport and maturation. The uptake of alpha factor pheromone by wild-type and Ypt7p-deficient cells was found to be indistinguishable, but in mutant cells lacking Ypt7p, degradation of the endocytosed pheromone was severely inhibited. Our findings suggest a role of Ypt7p in protein transport between endosome-like compartments.


YDJ1p facilitates polypeptide translocation across different intracellular membranes by a conserved mechanism.

The role of S. cerevisiae YDJ1 protein (YDJ1p) in polypeptide translocation across membranes has been examined. A conditional ydj1 mutant strain (ydj1-151TS) is defective for import of several polypeptides into mitochondria and alpha factor into the endoplasmic reticulum at 37 degrees C. These defects are suppressed by E. coli dnaJ or overexpression of S. cerevisiae SIS1 proteins. A different ydj1 mutant, which cannot be farnesylated (ydj1-S406), displays similar transport defects to the ydj1-151 strain. Furthermore, the ability of purified ydj1-151p to stimulate the ATPase activity of hsp70SSA1 was greatly diminished compared with the wild-type protein. Together, these data suggest that YDJ1p functions in polypeptide translocation in a conserved manner, probably acting at organelle membranes and in association with hsp70 proteins.


The cytoplasmic carboxy-terminal amino acid specifies cleavage of membrane TGF alpha into soluble growth factor.

Membrane-anchored transforming growth factor alpha (proTGF alpha) belongs to a group of transmembrane proteins whose extracellular domains are selectively cleaved and released into the medium. We demonstrate that the carboxy-terminal valine in the cytoplasmic tail of proTGF alpha is required for cleavage of the growth factor ectodomain in response to various activators. This cleavage process occurs outside Golgi or lysosomal locations, affects cell surface proTGF alpha, and requires little or no membrane traffic. We propose that cleavage and release of proTGF alpha ectodomain involve a specialized proteolytic system and depend on the recognition of a simple and specific determinant located in the proTGF alpha cytoplasmic tail.


Molecular characterization of the mouse agouti locus.

The agouti (a) locus acts within the microenvironment of the hair follicle to regulate coat color pigmentation in the mouse. We have characterized a gene encoding a novel 131 amino acid protein that we propose is the one gene associated with the agouti locus. This gene is normally expressed in a manner consistent with a locus function, and, more importantly, its structure and expression are affected by a number of representative alleles in the agouti dominance hierarchy. In addition, we found that the pleiotropic effects associated with the lethal yellow (Ay) mutation, which include pronounced obesity, diabetes, and the development of neoplasms, are accompanied by deregulated overexpression of the agouti gene in numerous tissues of the adult animal.


TAP1 mutant mice are deficient in antigen presentation, surface class I molecules, and CD4-8+ T cells.

The transporter associated with the antigen processing 1 (TAP1) gene encodes a subunit for a transporter, presumed to be involved in the delivery of peptides across the endoplasmic reticulum membrane to class I molecules. We have generated mice with a disrupted TAP1 gene using embryonic stem cell technology. TAP1-deficient mice are defective in the stable assembly and intracellular transport of class I molecules and consequently show severely reduced levels of surface class I molecules. These properties are strikingly similar to those described for the TAP2 mutant cell line RMA-S. Cells from the TAP1-deficient mice are unable to present cytosolic antigens to class I-restricted cytotoxic T cells. As predicted from the near absence of class I surface expression, TAP1-deficient mice lack CD4-8+ T cells.


The GCN4 basic region leucine zipper binds DNA as a dimer of uninterrupted alpha helices: crystal structure of the protein-DNA complex.

The yeast transcriptional activator GCN4 is 1 of over 30 identified eukaryotic proteins containing the basic region leucine zipper (bZIP) DNA-binding motif. We have determined the crystal structure of the GCN4 bZIP element complexed with DNA at 2.9 A resolution. The bZIP dimer is a pair of continuous alpha helices that form a parallel coiled coil over their carboxy-terminal 30 residues and gradually diverge toward their amino termini to pass through the major groove of the DNA-binding site. The coiled-coil dimerization interface is oriented almost perpendicular to the DNA axis, giving the complex the appearance of the letter T. There are no kinks or sharp bends in either bZIP monomer. Numerous contacts to DNA bases and phosphate oxygens are made by basic region residues that are conserved in the bZIP protein family. The details of the bZIP dimer interaction with DNA can explain recognition of the AP-1 site by the GCN4 protein.


Enhancement of the growth of human osteosarcoma cells by human interferon-gamma.

It is well known that interferons inhibit cell growth. However, we found that human interferon-gamma (HuIFN-gamma) enhanced the growth of human osteosarcoma cells, HOS-Y1 cells, in a dose-dependent manner. This enhancing effect was found only under the following conditions: when the cells were precultured for 2 or 3 days and then treated with HuIFN-gamma for 2, 3, or 4 days, and when the cells were seeded at a density of 1,000 or 2,000 cells/well. The degree of enhancement of cell growth was maximum when the cells were precultured at a density of 1,000 cells/well for 3 days and then treated with HuIFN-gamma for 2 days. The enhancing effect of HuIFN-gamma disappeared in the presence of anti-HuIFN-gamma antibody. In addition, it was found that the conditioned medium from HOS-Y1 cells enhanced the growth of HOS-Y1 cells, and that the conditioned medium from HOS-Y1 cells cultured with HuIFN-gamma enhanced the cell growth more than that from cells cultured without HuIFN-gamma. Epidermal growth factor (EGF), acidic fibroblast growth factor (aFGF), basic fibroblast growth factor (bFGF), and transforming growth factor-beta 1(TGF-beta 1) did not enhance the growth of HOS-Y1 cells. These results suggest that HuIFN-gamma enhanced the cell growth by augmenting the production of unknown growth factor(s) in HOS-Y1 cells via an autocrine mechanism.


Activation of apoalkaline phosphatase by serum albumin with Zn2+ in rat hepatoma cells.

Reuber rat hepatoma cells (R-Y121B) cultured at 0.5% serum accumulated apoalkaline phosphatase in intact cells. When R-Y121B cells were cultured in the presence of bovine serum albumin, alkaline phosphatase activity increased in the cells, and the associated increase in enzyme activity differed amongst bovine serum albumin preparations. The treatment of bovine serum albumin with activated charcoal not only enhanced the effect of serum albumin on alkaline phosphatase activity, but also cancelled the differences due to different preparations of serum albumin. In contrast, no effect from serum albumin was observed in the increase of alkaline phosphatase activity in R-Y121B cell homogenates incubated at 37 degrees C. The activated-charcoal treatment of bovine serum albumin increased the amount of Zn2+ bound to the protein. When R-Y121B cells were cultured with bovine serum albumin, the concentration of Zn2+ in the cytosol fraction slightly increased. However, the effect of serum albumin on Zn2+ concentration in the cytosol fractions was independent of charcoal treatment. It was concluded that serum albumin with Zn2+ induces the activation of apoalkaline phosphatase due to Zn2+ binding.


Ultrastructure of detergent-resistant cytoskeletons in the noncortical domain of sea urchin eggs as revealed by the quick-freeze deep-etch technique.

The ultrastructure of detergent-resistant cytoskeletons in the noncortical cytoplasm of sea urchin eggs was studied by quick-freeze, deep-etch electron microscopy. Two different cytoskeletal organizations were identified in the detergent-treated sea urchin eggs. They were distinguished by the presence or the absence of long actin filaments and probably correspond to the cortex and the noncortical cytoplasm, respectively. The non-cortical cytoplasm was composed of a complex network (designated here as the ground network) of filaments 6 to 13 nm in diameter, that interconnected aggregates of small globular materials, yolk granules and a meshwork of uniform filaments (8-9 nm in diameter). The 6 to 13 nm filaments comprising the ground network were branched and associated with filaments of the same or other sizes, resulting in the formation of an extremely complex network. The meshwork of 8-9 nm filaments was homogeneous in composition and constitutes a novel structure which has not been previously described. The 8-9 nm filaments were connected to one another at their ends, forming a meshwork of polygons. Meshworks, ranging up to 3 microns in diameter, were distributed throughout the non-cortical cytoplasm of the egg. Similar cytoplasmic structures were also observed in fertilized eggs.


Immunocytochemical study of recombinant terminal deoxynucleotidyl transferase (TdT) synthesized by baculovirus-infected insect cells.

The ability to overproduce terminal transferase through recombinant DNA technology should provide alternate means for generating sufficient quantities for structural and mechanistic study of this creative DNA polymerase. In this work we have investigated, at electron microscope level, the morphological modification and ultrastructural localization of synthesized human terminal transferase occurring in Sf-9 cells during recombinant baculovirus infection time. The results obtained showed that TdT is localized and stored only at the cytoplasmic level; the nucleus did not show any specific site able to link the neosynthesized TdT. The amount of the enzyme, estimate by immunostaining analysis, increased with the viral infection time. Morphological changes occurring during viral infection consist mainly of variations of cellular surface, different size and shape of cytoplasmic organelles and modification of nuclear components.


Effects of fibronectin-type V collagen recombinant fusion protein on cell adhesion and cell proliferation.

An expression vector pTF7520-Col-V-In, which encodes a fusion protein of the cell-binding domain of fibronectin (C277) and the insulin- and heparin-binding domain of the alpha 1 chain of human type V collagen, was constructed. E. coli transfected with this plasmid synthesized a 50-kDa fusion protein. This fusion protein, C277-V, was purified from the crude extract by a single step heparin HPLC. Similar amounts of insulin bound to purified C277-V and to the alpha 1 chain of type V collagen as judged by the binding of peroxidase-conjugated insulin. Cell-adhesive activity of C277-V was lower than that of the original fibronectin fragment C274, but similar numbers of cells adhered to both protein substrates when the culture dishes were coated with 1 mM of each protein. Insulin bound to the C277-V substratum stimulated the growth of mouse mammary tumor MTD cells in serum-free culture medium.


Expression of a 66-kD heat shock protein associated with the process of cyst formation of a true slime mold, Physarum polycephalum.

Under unfavorable conditions for growth, haploid myxoamoebae of Physarum polycephalum retracted their pseudopodia and changed their cell shape into disk-like form, after which they constructed the cell walls to form microcysts. These morphological changes of haploid cells were associated with changes in intracellular distribution of actin filaments. Staining with phalloidin showed that actin filaments were almost uniformly distributed throughout the cytoplasm of the myxoamoebae. When these cells were transferred to a cyst-inducing medium, the actin structures changed into short rods or dots, after which the rods/dots disappeared in the microcysts. An incubation of the myxoamoebae in the cyst-inducing medium caused the synthesis of several proteins, among which a 66-kD protein was most prominently induced. The morphological changes and the induction of the 66-kD protein was pronounced at elevated temperatures, e.g. 40 degrees C. The 66-kD protein was not induced, however, when plasmodia of the same species were incubated at 40 degrees C. We found that the 66-kD protein was co-precipitated with polymerized actin and bound to ATP-agarose. A double staining of the disk-shaped cells with anti-66-kD protein antibody and phalloidin revealed superimposable localization of the 66-kD protein and actin filaments in the short rods or dots. Although the induction of the 66-kD protein was enhanced at high temperatures, the protein was immunologically unrelated to the common heat shock proteins, HSP70 and HSP90, those are highly conserved during evolution. These results indicate that the 66-kD protein is a novel heat shock protein which is specifically expressed during cyst formation.


Penetration of Toxoplasma gondii into host cells induces changes in the distribution of the mitochondria and the endoplasmic reticulum.

Fluorescence microscopy, using dyes which specifically label mitochondria, endoplasmic reticulum and the Golgi complex, and transmission electron microscopy, were used to analyze the changes which occur in the organization of these structures during interaction of Toxoplasma gondii with host cells. In uninfected cells the mitochondria are long filamentous structures which radiate from the nuclear region toward the cell periphery. After parasite penetration they become shorter and tend to concentrate around the parasite-containing vacuole (parasitophorous vacuole) located in the cytoplasm of the host cell. The mitochondria of extracellular parasites, but not of those located within the parasitophorous vacuole, were also stained by rhodamine 123. Labeling with DiOC6, which binds to elements of the endoplasmic reticulum, in association with transmission electron microscopy, revealed a concentration of this structure around the parasitophorous vacuole. The membrane lining this vacuole was also stained, suggesting that components of the endoplasmic reticulum are also incorporated into this membrane. The Golgi complex, as revealed by staining with NBD-ceramide and electron microscopy, maintains its perinuclear position throughout the evolution of the intracellular parasitism.


Relationship between direction of rolling and yawing of golden hamster and sea urchin spermatozoa.

As a first step towards understanding the function and mechanism of spiral movement of spermatozoa swimming through a medium, the direction of rolling (rotational movement of the spermatozoa around their long axis) and that of yawing (circular motion of spermatozoa upon the surface of a glass microscope slide and coverslip) were examined for golden hamster and sea urchin spermatozoa. Most golden hamster spermatozoa yawed clockwise over the upper surface of a glass slide when viewed from above, whereas in most sea urchin spermatozoa yawing was counterclockwise. Under the lower surface of a coverslip, the direction of yaw of golden hamster or of sea urchin spermatozoa was reversed. Most golden hamster spermatozoa rolled counterclockwise as seen from the anterior end, whereas all examined sea urchin spermatozoa rolled clockwise relative to the observer. On the basis of quantitative analysis of the proportion of spermatozoa rolling (or yawing) clockwise to those rolling (or yawing) counterclockwise, a close relationship between the direction of rolling motion and that of yawing motion was shown for both golden hamster and sea urchin spermatozoa.


[Gestrinone in the therapy of sterility due to endometriosis].

Thirty-two infertile women with laparoscopic diagnosis of asymptomatic pelvic endometriosis were treated with gestrinone 2.5 mg twice weekly for 6 months. Subsequently, a second-look laparoscopy was performed in all of them. All of the patients were followed for at least 12 months after the end of treatment, during which time they attempted to conceive. Ten (31.25%) from all 32 treated women became pregnant. The highest incidence of pregnancy was in the stage I (12.5%) and II (12.5%) (rAFS) of endometriosis, a half of them in stage III (6.25%) and no pregnancy was recorded in the stage IV found prior to the beginning of therapy. There was no statistically significant (chi 2-test) relationship between the reduction of laparoscopic findings (defined as a visual extermination or as a decrease in staging of endometriosis) and pregnancy. In 19 (59.4%) from 32 treated women the stage of endometriosis remained unchanged, in 9 (28.3%) it was reduced by one up to three stages and in 4 (12.5%) the endometriosis was visually eliminated. In no case the staging of endometriosis was worsened during therapy. Gestrinone was well tolerated. In no one of patients it was necessary to interrupt the treatment untimely.


[Lynestrenol and allylestrenol in the therapy of postmenopausal hot flushes].

In a parallel randomized placebo controlled clinical trial the authors tested two synthetic gestagens--allylestrenol and lynestrenol--in the treatment of postmenopausal flushes. Both preparations were administered during a six-week period in rapidly declining doses. Allylestrenol was administered in initial doses of 30 mg/d, after five days the doses were reduced so that after 15 days a daily dose of 5 mg was reached. The initial dose of lynestrenol was 10 mg/d with a similar gradual decline to 1.25 mg per day. The trial comprised 42 women with menopausal flushes after a natural or artificial menopause (castration). Both preparations improved the subjective condition of the patients and reduced the gonadotropic production significantly better than placebo (p < 0.05). Subjective relief after lynestrenol was recorded after the second week of therapy, in allylestrenol only after the sixth week when the effects of the two preparations were equal and significantly better than after placebo. Suppression of gonadotropin production was similar after both preparation but with a more rapid onset after lynestrenol and a more prolonged effect after allylestrenol even after significant reduction of the doses. No serious undesirable effects were recorded, no changes in indicators of liver functions or serum lipids incl. the HDL/LDL ratio. Endometrial bleeding after administration of hormones was not more frequent than after placebo. In the discussion the authors analyze some aspect of treatment of the climacteric syndrome by means of these hormones. Allylestrenol in particular is an interesting gestagen due to its inherent oestrogenic effect in the absence of an androgenic effect.


[Anti-sperm antibodies in women and men in infertile marriages].

The authors examined the serum of 76 women and 33 men from infertile marriages for the presence of antispermatozoal antibodies. Seropositivity was revealed in 19 women (25%) and in 12 men (36%). In control groups of healthy men and women seropositivity was substantially less frequent. In women the difference was statistically significant, but not in men. The ELISA method which was applied for examination appears to be a useful tool for screening of immunological factors of infertility, in particular in couples who where infertile for prolonged periods and where no unequivocal cause of infertility was found. Only more detailed comprehensive examination can explain immunologically conditioned cases of infertility and lead to selection of an optimal therapeutic strategy.


[Immunoreactivity of Mycoplasma hominis PG21 antigens and human sperm antigens].

The authors selected from the WHO bank for reproductive immunology ten sera containing concurrently antispermatozoal antibodies and antibodies against Mycoplasma hominis PG21. By absorption examination of selective sera with a different concentration (10(9)/ml, 3.10(8)/ml, 10(8)/ml) of human spermatozoa and a rinsed sonicate of Mycoplasma hominis PG21 in dilutions of 1:20, 1:80, 1:320 the authors did not find a crossed immunoreactivity between sperm antigens and mycoplasmic antigens.


Mechanism of resistance to sulphite in Saccharomyces cerevisiae.

Growth inhibition and cell killing caused by sulphite were reduced in seven Saccharomyces cerevisiae sulphite-resistant independent mutants, compared to their parental strains. Genetic analysis showed that in the seven mutants resistance was inherited as a single-gene dominant mutation and that all the analyzed mutations were allelic, thus identifying a major gene responsible for sulphite resistance in S. cerevisiae. Two of the mutants, MBS20-9 and MBS30, were further characterized. 35S-sulphite uptake experiments showed that the ability to accumulate sulphite was markedly reduced in the two resistant strains. No difference between resistant and sensitive strains with respect to glyceraldehyde-3-phosphate dehydrogenase sensitivity to sulphite, or to intracellular glutathione content, were revealed. In contrast, the extracellular acetaldehyde concentration was higher in the resistant mutants, both in the presence and in the absence of sulphite.


Genetic diversity of the yeast Candida utilis.

The electrophoretic karyotypes and some mtDNA restriction fragment patterns of 13 strains of Candida utilis and one strain of Hansenula jadinii were compared. PFGE separations revealed remarkable chromosome length polymorphisms between two groups of strains suggesting that perhaps they do not belong to the same species. However, all strains had the same or similar EcoRI, HindIII and BamHI mtDNA restriction patterns. The mtDNA genomes had an average size range of 55 kb. These results support the supposition that C. utilis is a yeast with a highly variable electrophoretic karyotype as already known for another imperfect yeast species, Candida albicans.


Sequence analysis of the glyceraldehyde-3-phosphate dehydrogenase genes from the basidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus.

GPD genes encoding glyceraldehyde-3-phosphate dehydrogenase were isolated from the homobasidiomycetes Schizophyllum commune, Phanerochaete chrysosporium and Agaricus bisporus. All three species contain one transcriptionally active GPD gene, but A. bisporus also contains an inactive GPD gene (tandemly linked to the active gene). These genes contain 5-9 introns located at conserved positions, differing (except in one case) from intron positions in ascomycetous GPD genes. The predicted amino-acid sequences of the proteins encoded by the three active GPD genes are highly homologous. A comparison with protein sequences from filamentous ascomycetes shows a clear distinction, whereas the GPD genes from ascomycetous yeasts are quite distinct from both the filamentous ascomycetes and basidiomycetes. Promoter regions of ascomycetous GPD genes do not correspond to those of the GPD genes of basidiomycetes which may (partly) explain poor expression in basidiomycetes of introduced genes driven by an ascomycete GPD promoter.


The 5S ribosomal RNA gene is linked to large and small subunit ribosomal RNA genes in the oomycetes, Phytophthora vignae, P. cinnamomi, P. megasperma f.sp. glycinea and Saprolegnia ferax.

Southern hybridization and polymerase chain reaction data indicate that the 5S ribosomal RNA gene is linked to the ribosomal RNA gene repeat unit in the oomycetes, Phytophthora vignae, P. cinnamomi, P. megasperma f.sp. glycinea and Saprolegnia ferax, and is apparently transcribed in the same direction as the large and small subunit ribosomal RNA genes. The polymerase chain reaction has been used to amplify all components of the entire ribosomal RNA gene repeat unit for each of these oomycetes. The total size of all amplified products is identical to the size of the ribosomal RNA gene repeat unit, as determined by Southern analysis.


Mitochondrial DNA loss by yeast reentry-mutant cells conditionally unable to proliferate from stationary phase.

Double-mutant cells of the budding yeast Saccharomyces cerevisiae harboring the gcs1-1 and sed1-1 mutations are conditionally defective (cold-sensitive) only for reentry into the mitotic cycle from stationary phase. If already proliferating at the permissive temperature (29 degrees C), these reentry-mutant cells continue to proliferate when transferred to the restrictive temperature of 14 degrees C, but under these conditions reentry-mutant cells lose mitochondrial DNA (mtDNA). In addition, upon exhaustion of the nutrient supply at 14 degrees C, these reentry-mutant cells entered stationary phase at a decreased cell concentration and did not accumulate the reserve carbohydrates trehalose and glycogen. Both of these deficiencies were due to the loss of mtDNA, as shown by the responses of wild-type cells also lacking mtDNA. Mitochondrial status did not affect other aspects of the reentry-mutant phenotype. Although mitochondrial activity and the accumulation of carbohydrate reserves are typical features of cells in stationary phase, the reentry-mutant phenotype reveals that neither entry into nor exit from stationary phase need involve mitochondrial function.


Evolutionary analysis of the plastid-encoded gene for the alpha subunit of the DNA-dependent RNA polymerase of Pyrenomonas salina (Cryptophyceae).

The nucleotide sequence of the gene coding for the plastid-encoded alpha subunit of DNA-dependent RNA polymerase from the cryptomonad alga Pyrenomonas salina was determined. The deduced amino-acid sequence, corresponding to a 35.2 kDa polypeptide, was compared to homologues from other organisms. Evolutionary relationships were analyzed in detail by the parsimony method together with bootstrap analysis. The deduced phylogenetic tree shows that the cryptomonad gene is the most ancient type of known plastid-encoded RNA polymerase.


The influence of the Ustilago maydis REC1 gene on plasmid-chromosome recombination.

The REC1 gene of U. maydis has an important but ill-defined role in DNA recombination and repair. We have examined its role in plasmid-chromosome recombination. Plasmid DNA was linearized at various locations with respect to the cloned U. maydis PYR3 gene and introduced into cells by transformation. Chromosomal integration and repair by an homologous cross-over with plasmid containing a double-strand break or gap in the PYR3 gene was markedly reduced in the absence of the REC1 gene product. Homologous replacement of the chromosomal pyr3-1 allele by a single copy of wild-type sequences from plasmid cut outside PYR3 was not found in the absence of the REC1 product. Instead, novel transformants generated in its absence suggests that ligation plays a role in their generation.


The linear mitochondrial plasmid pAL2-1 of a long-lived Podospora anserina mutant is an invertron encoding a DNA and RNA polymerase.

The molecular characterization of an additional DNA species (pAL2-1) which was identified previously in a long-lived extrachromosomal mutant (AL2) of Podospora anserina revealed that this element is a mitochondrial linear plasmid. pAL2-1 is absent from the corresponding wild-type strain, has a size of 8395 bp and contains perfect long terminal inverted repeats (TIRs) of 975 bp. Exonuclease digestion experiments indicated that proteins are covalently bound at the 5' termini of the plasmid. Two long, non-overlapping open reading frames, ORF1 (3,594 bp) and ORF2 (2847 bp), have been identified, which are located on opposite strands and potentially encode a DNA and an RNA polymerase, respectively. The ORF1-encoded polypeptide contains three conserved regions which may be responsible for a 3'-5' exonuclease activity and the typical consensus sequences for DNA polymerases of the D type. In addition, an amino-acid sequence motif (YSRLRT), recently shown to be conserved in terminal proteins from various bacteriophages, has been identified in the amino-terminal part of the putative protein. According to these properties, this first linear plasmid identified in P. anserina shares all characteristics with invertrons, a group of linear mobile genetic elements.