Assessment of a strategy for the control of respiratory diseases in children. A programme for the control of respiratory diseases in children was conceived for the State of S. Paulo, Brazil, in 1986. Its progress thereafter and the epidemiology of the diseases concerned are examined. Apart from an inquiry into the 64 existing State local health authorities, a sample of 18,255 cases of children assisted by the programme at different levels, including both in-patient and outpatient care, is analysed. Each case record included information about identification (child, doctor and health facility), reasons for calling, diagnoses made and outcome of treatment. Further data were also sought from hospitals and from State mortality records. The programme was found to be poorly implemented in the State but, where implemented, it showed itself capable of resolving problems (only 0.5% of the cases could not be handled) as also of changing ongoing trends (more than 50% reduction in hospital admission rates). Individual assessment of each item of the programme indicated its bottlenecks. Regarding the epidemiology of respiratory diseases, it is observed that the major burden to health services comes from children aged less than five, and that the most important diseases are wheezing illnesses and pneumonia. Moreover, they were found to be significantly associated (p = 0.000) so that a child in the community presenting wheezing diseases is 5 times more likely to develop pneumonia than a child with any other respiratory diagnosis. Similarly, among the under five deaths it was found that the risk for pneumonia is 3 times greater for children who died presenting wheezing diseases than it is for children with any other sort of diagnosis. In conclusion, the programme is deemed to be efficient and effective but its efficacy is marred by administrative flaws. The successful control of respiratory problems in childhood is related to a proper appreciation of the importance of wheezing diseases.


Geographical patterns of proportionate mortality for the most common causes of death in Brazil. Mortality due to chronic diseases has been increasing in all regions of Brazil with corresponding decreases in mortality from infectious diseases. The geographical variation in proportionate mortality for chronic diseases for 17 Brazilian state capitals for the year 1985 and their association with socio-economic variables and infectious disease was studied. Calculations were made of correlation coefficients of proportionate mortality for adults of 30 years or above due to ischaemic heart disease, stroke and cancer of the lung, the breast and stomach with 3 socio-economic variables, race, and mortality due to infectious disease. Linear regression analysis included as independent variables the % of illiteracy, % of whites, % of houses with piped water, mean income, age group, sex, and % of deaths caused by infectious disease. The dependent variables were the % of deaths due to each one of the chronic diseases studied by age-sex group. Chronic diseases were an important cause of death in all regions of Brazil. Ischaemic heart diseases, stroke and malignant neoplasms accounted for more than 34% of the mortality in each of the 17 capitals studied. Proportionate cause-specific mortality varied markedly among state capitals. Ranges were 6.3-19.5% for ischaemic heart diseases, 8.3-25.4% for stroke, 2.3-10.4% for infections and 12.2-21.5% for malignant neoplasm. Infectious disease mortality had the highest (p < 0.001) correlation with all the four socio-economic variables studied and ischaemic heart disease showed the second highest correlation (p < 0.05). Higher socio-economic level was related to a lower % of infectious diseases and a higher % of ischaemic heart diseases.(ABSTRACT TRUNCATED AT 250 WORDS)


[Body mass index (body mass.body height-2) as indicator of nutritional status in adults: review of the literature]. The limitations and possible uses of the body mass index (BMI) or Quételet index, calculated as the ratio of body mass (kg), and squared stature (m), as an indicator of the nutritional status of adults, are reviewed. The cut-off values for overweight (BMI > or = 25) and chronic energy deficiency (BMI < 18.5) are discussed in the light of the available international literature. Even though BMI does not assess the body composition (BC) of individuals, the case with which it is measured is a strong argument for the use of BMI in epidemiological studies associated otherwise with other anthropometric measurement until such a time as simpler field methods of the assessment of BC are developed.


[Factors associated with the non-utilization of contraceptives in adolescence]. The literature concerning the major correlates of the non-utilization contraceptives in adolescence is critically reviewed. Research findings are analyzed and discussed in terms of their contribution not only to a deeper understanding of adolescents' risk-taking sexual behavior, but also to the implementation of more effective and realistic sexual education for this age-group.


[Ectopic pregnancy--an analysis of the cases treated at the N. I. Pirogov Research Institute of Emergency Medicine]. Rate of ectopic pregnancy, a life-threatening pathology with great psycho-social significance, had reached from 4.5 per miles in 1970 up to 16.8 per miles in 1987. During a nine-year period (1980-1988) 466 women with diagnosis ectopic pregnancy were hospitalized in the Department of Urgent Gynaecology-Emergent Medical Institute "Pirogov". Married were 285 (61.16%) of them, not married--116 (24.89%) and divorced--65 (13.95%). Dull, not localized pain was observed in 300 (64.38%) of them. In 160 (34.33%) only colics were observed, dated from 15 days to 1 month before hospitalization. The classic triad of amenorrhea, colicky pains in the lower abdomen and bloody liquids, running out of genitalia. When hospitalized, 194 women (41.63%) were in very severe general condition, from these 92 in severe hemorrhagic shock. The blood loss, evaluated in the beginning of the operation was found to be: 0 ml--8 patients (1.72%); up to 300 ml--143 (30.69%); up to 600 ml--121 women (25.97%); up to 1000 ml--102 (21.88%); up to 1500 ml--36 (7.73%); up to 2000 ml--36 patients (7.73%); up to 3000 ml--17 (3.65%); more than 3000 ml--3 (0.64%). A conclusion was made that it is necessary to improve the prophylaxis, the diagnostic methods and the treatment of ectopic pregnancy, so as to save reproductive abilities of the women.


Oligospermia associated with normal testicular function and epididymal lesions or malpositions. Epididymal disease is in our experience responsible for about 40 per cent of male infertility situations. The diagnosis of epididymal oligospermia can be suspected by the presence of firm normal volume testicles, by the absence of any elevation of FSH, and eventually, by a (slightly) hardened epididymis at palpation. The definite diagnosis however will only be established by quantitative reading of the testicular biopsy. Besides in oligospermic patients clinical information isolates an eventual new group where healthy epididymis could interfere with sperm quality by its malposition.


[A program for the antenatal diagnosis and management of fetal retardation at the Obstetrical-Gynecological Clinic of the Medical University of Pleven (preliminary report)]. The goal of this preliminary report is to define the principal stages of a program for antenatal diagnosis and management in cases of IUGR that has been introduced in practice in the Department of Obstetrics and Gynaecology at the Medical University--Pleven. We present a modified scoring system after A. Wennergren and K. Karlsson (1982) that was distributed to all primary obstetrics care phases of the city and the region for screening of all pregnancies at risk for IUGR. alpha-FP levels in the amniotic fluid and in maternal blood serum are followed according to the indication. Ultrasound follow-up of 8 biometric parameters of the foetus is performed at 2 weeks intervals. The scoring system for foetal biophysical profile after A. Vintzileos (1983) in combination with the AF indexers is introduced as a routine practice. Foetal lung maturation is surveyed by quantitative obtaining of lecithin/sphingomyelin ration in the AF after Gluck (1974). Decision on way and time of delivery is made based on the complex results.


[Cardiotocographic control during labor]. The authors discuss different aspects of intrapartum foetal monitoring. The scientific basis of foetal intrapartum foetal monitoring are pointed out. The authors present three groups of cardiotocographic signs which should alarm the obstetricians: warning signs, threatening signs and clear signs for foetal distress. The authors discuss also the specific characteristics of the cardiotocographic record of premature fetus during pregnancy and labour. It is pointed out that the intrapartum foetal monitoring and the correct interpretation of the cardiotocographic record lead to better perinatal results.


Towards the definition of the fertilizing spermatozoa. The seminological work-up of sperm samples is today totally different than only 20 years ago and the new more detailed readings bring the seminologist closer to the definition of the fertilizing capacity of sperm samples. Still, however careful the descriptive semen analysis, in 10% the predictive value is wrong. In view of inadequacies of the descriptive semen analysis, attempts to improve the laboratory contributions have focussed on the development of more refined techniques concerning the functional competence of human spermatozoa: computerized study of sperm motility, zona-free hamster oocyte penetration assay, hemizona-binding assay, study of the acrosoma reaction, biochemical studies. But it must be considered that the cost of all sperm controls in order to evaluate its fertilizing capacity can become on even harder financial burden than on IVF attempt itself.


[Episiotomy repair by the method of W. B. Shute]. For about 1 year 106 episiotomy are reapproximated using figure-of-eight sutures by W. B. Shute and 623 by using conventional techniques. Early (4-6 days) and lately (6 months and more) results are compared in these 2 groups. Authors have found out benefits of Shute's technique and recommend to use it as a routine procedure in current obstetric practice.


Assisted reproductive technologies in severe male infertility. The increasing incidence of male subfertility as an indication for ART is first discussed. The tendency to use assisted reproduction techniques in such cases is attributed to the disappointing results of classical treatments. The Authors deal with two problems: the choice of the best treatment methods of the sperm and the choice of the best technique of ART for treating male infertility. The analysis of 138 couples treated for male subfertility showed that the centrifugation on discontinuous Percoll gradients (CDPG) and especially on the mini-Percoll (mini CDPG) offers the best results if compared with pellet swim up and other techniques. With reference to the technique of choice, an accurate analysis of tubal (TET and ZIFT) and uterine (IVF/ET) transfers shows that no advantage seems to be obtained with the more sophisticated and exacting tubal transfer. This final conclusion is presently evaluated on the basis of a retrospective study.


[A case of vaginal delivery--11 months after a primary cesarean section]. The state of the uterine cicatrix and more specifically its sufficiency are extremely important for the obstetrician when he has to choose between a second caesarean operation and a vaginal delivery. This decision determines the post partum and postoperative techniques and behaviour. The author reports a case of a woman who has vaginally delivered a child 11 months after a caesarean operation. This case demonstrates that approaching personally the obstetrician can decide in favour of a vaginal delivery after a previous caesarean operation even in the cases of pregnancy which has begun before the 6 month period after the caesarean operation has elapsed.


Spermiophages in human semen in normal subjects and in patients with selected diseases. In order to screen for spermiophages in human semen, a Neutral Red Test was carried out in: a) normal subjects (NORMAL); b) male partners of infertile couples (INFERT); c) subjects with selected diseases. Spermiophages were observed only in 7.5% of the normospermic subjects, while the test was positive in 26.5% of INFERT. Positivity was higher in the selected pathological groups, even compared to the INFERT group. This observed increase in spermiophage activity could be an important pathogenetic factor in some cases of male infertility.


[The age of women who deliver vaginally after a prior cesarean section]. The author has studied in detail the structure of age groups of women delivered vaginally after a previous caesarean section over a period of ten years. He supports the view that the combination of an "aged lying-in woman" (over 29) and a previous caesarean section should not be in itself an obligatory indication for a planned caesarean section as it is considered now. So, the conclusion is that the indication "aged primipara" (over 29) as an only indication for a planned caesarean section should be doubted since 70 women over the age of 29 and after a previous caesarean section delivered vaginally without any complications for the mother or fetus.


[Our experience in the diagnosis of chlamydial cervicitis with the Chlamydiasyme and Chlamyset Antigen tests]. The authors analyse in the present study the results of the examination of 309 female patients for chlamydial infection, using Chlamyset Antigen (Orion Diagnostica--Finland) and Chlamydiasyme Diagnostic Kit (Abbott Labs--USA). Chlamydial antigen was found in the endocervix of 15.2% of the examined cases. Comparing the enzyme immunomethod Chlamydiasyme with the immunofluorescent method Chlamyset Antigen the authors found 94.4% diagnostic accuracy.


[The use of low-energy lasers via action on the acupuncture points in inflammatory processes in the adnexa]. The paper deals with the effect of low-energy lasers in treating cases of subacute and chronic inflammatory processes of the adnexa through impact on biologically active points. The results in 30 patients treated with laser puncture have been observed and compared with the results in 30 women treated according to the classical methods. It has been found that the patients with subacute and chronic inflammatory processes treated with laser puncture spend in hospital 3 days less than the other patients and the pain symptoms disappear quickly. It can be concluded that the application of low-energy lasers through impact on biologically active points in subacute and chronic adnexitis can find its place in gynaecological practice.


[The effect of E2 and progesterone on the receptor status in endometrial carcinoma]. The authors study the plasma levels of E2 and progesterone in 62 women with EC. Parallelly the ER and PR in the same number of cases have been studied. The values of E2 in the plasma of ER+ are much lower than those of E2 in ER-. The quantity of E2 and progesterone influence the values of ER and PR in cancers of women in pre and postmenopausal age. In positive ER and PR patients the level of E2 and progesterone is lower in comparison of ER- and PR-.


[The posttherapy survival of patients with invasive cancer of the vulva]. The treatment results and five-years survival are studied in 157 patients with invasive vulvar cancer treated for the period of 1979-1988 in Gynaecological Clinic of the National Oncological Center Institute in Sofia. The general five years survival is 59.2%. The highest is the survival of the patients with first clinical stage--79.55%. The best treatment results are by patients treated with radical vulvectomy and regional lymph node dissection, without histological founded metastases in the extirpated lymph nodes--69.24% five years survival. The authors show some better results because of the better early diagnosis of the disease and the improved therapeutical methods.


[Extrauterine pregnancy--the clinical experience of using a new differential-diagnostic immunological test]. The authors report results of immunologic test, that has been used to help the decision in some difficult for differential diagnosis cases. Two immunologic parameters were investigated--the amount of circulating immune complexes (CIC) and the lymphocytic nuclear coefficient (LNC). The results are put on a specially designed nomogram. The combined test used in the Department of "Obstetrics and Gynaecology"--Pleven, gives excellent results. The advantages of the test are discussed and a clinical case is presented as an illustration.


[Estrogen and progesterone receptors as prognostic markers in endometrial carcinoma]. The authors examined the concentration of ER and PR in malignant endometrium and compared them to the survival of the patients with endometrial carcinoma. The results were 55.5% positive ER and 44.5% negative, PR-74.1% positive and 25.9% negative. From 54 women with EC-7 died during 3 years period of time. From these 7, only one was with elevated concentrations of both ER and PR. 5 women were with negative PR and in 3-negative both ER and PR. It's considered that negative PR are the most important prognostic signs for the survival of patients with EC.


[The lead concentration of the maternal serum in cases of missed abortion]. The neurotoxic effect of lead is well known, but the investigations of the pregnant women are still few. The aim of the present study is to establish whether there is difference in the concentration of lead in the blood of the pregnant women with missed abortion in comparison with women with normally developing pregnancy in the same gestational age. There were examined 35 patients with missed abortion and a control group of 26 women with normal pregnancy in the same gestational age. There were established statistically significant higher levels of lead (p < 0.001) in women with normal pregnancy (124.46 +/- 46.65 micrograms/l) in comparison with cases with missed abortion (183.73 +/- 24.16 micrograms/l). There were discussed the possible mechanisms, which can be responsible for the observed changes.


[The motivation for deciding to preserve or to terminate a pregnancy]. With 68 women recently confined and terminating pregnancy by wish a study was made about anxiety in its two varieties: (i) as a mental condition connected with an actual and significant for the person life situation and (ii) as a personal feature--a way of perception and interpretation of the events as threatening. Anxiety's level is measured by a psychological test adapted to the Bulgarian conditions. The high average estimation of anxiety shows that termination of pregnancy is suffered as a stress. Subject to anxiety are also family's incomes when investigating both groups--women recently confined and women going to have an abortion. Level of anxiety of women recently confined is connected with the attitude towards future which definitely bothers 67.6 per cent of them. Analysis of indices shows that anxiety in connection with the decision for keeping or terminating of pregnancy as well as with the expected future after confinement is subject to many factors. Data interpretation confirms the well-known thesis that reproductive health protection is not only of a medical and biological but of very wide interdisciplinary interest when the woman is on the brink of the important for her personally and finally for the society as well decision pro and con real pregnancy.


[Psychological impact of cancer on the family: a review]. The impact of cancer on the patient's family and caregivers is important and it can be analyzed in terms of coping strategies. It depends on the patient (age, sex, role ...), the illness (site, stage, state ...), the people (parents, siblings, caregivers ...), the time (diagnosis, treatment, death, after death rehabilitation), the prognosis. The survey of the literature underlines the methodological difficulties met in this complex field and the distress of the family and caregivers at each stage of the illness, even during long term remissions.


["Alternative psychosis" illustrated by a clinical case]. We introduce here the "alternative psychosis" concept through the case of a 27 years old man, epileptic without psychiatrical background, who presented an acute 2 weeks long psychotical episode. We also call to mind the linkage, first described during the last century between epileptical disease and schizophrenic-like reactions. In 1934, Von Meduna noticed several times an improvement of patients affected by comitial crisis, when treating them by convulsive treatment. He was then the first to introduce the use of convulsive treatment for schizophrenic psychosis. In 1953, Landolt described the "forced normalisation" which is an improvement of the EEG recording of epileptical patients during psychotical episodes. Later on, Tellenbach introduced the term of "alternative psychosis", wishing thereby to insist on the clinical missing of epileptical crisis during those acute psychotical episodes, he also sharpens their symptomatical description.


[Psychiatry in the suburbs and suburbs of psychiatry]. A psychiatric service near Paris has opened two mobile units with the help and the support of local governments. The authors review their first year's activities involving preliminary consultation with patients without, in the first instance, prescribing treatment or giving any formal diagnosis. The team's action is resolutely preventive. The care of people at risk before they become really mentally ill. Psychiatry is brought from behind the closed doors of institutions to seek the cooperation of social workers and others in bringing about a better understanding of mental illnesses in local communities.


[Dysthymic schizophrenia. Reality or fiction?]. After an historical survey about the origin and the evolution of the concept of schizo-affective psychosis (i.e. in French schizophrénie dysthymique), a review of the recent bibliography reveals that the different models of classification are a subject for discussion. Studies tend to corroborate the model of an heterogeneous group with schizophrenic patients and manic-depressive patients.


[Elements of a neurophysiologic model of the unconscious]. The author sets forth a conception of an inborn reciprocally organized neuropsychological pre-program of the personality. In tender age it orientates the experience and the behaviour unconsciously and later its activity becomes conscious to one or another degree. Elements of this pre-program are a reciprocally organized system of the fundamental drives and emotions and adjacent structures. On the basis of this conception, the author makes a neuropsychological analyse of some para-adaptive personality reactions.


[Open and effectual scepticism of François Sanchez (1551-1623). Value for psychiatry]. "Quod Nihil Scitur" (That's that nothing we know) is a philosophical open, "adogmatic" and liberal form of scepticism. And also a model of methodology and epistemology for psychopathology and the history of psychiatry.


Elective mutism as a variant of social phobia. To examine the relationship between the syndromes of elective mutism and social phobia, a case of elective mutism associated with social phobia in a 12-year-old girl is presented, and the clinical literature regarding the syndrome of elective mutism is reviewed. Elective mutism or reluctance to speak in unfamiliar social situations may be a symptom of social phobia. Social anxiety is a nearly universal characteristic of children manifesting the syndrome of elective mutism. Elective mutism may respond to treatment with medications that also are effective in the treatment of social phobia. Elective mutism may be a manifestation of social phobia rather than a separate diagnostic syndrome. Pharmacologic treatment may be effective.


Cervicogenic dysfunction in muscle contraction headache and migraine: a descriptive study. The prevalence and nature of findings of cervicogenic dysfunction is explored in subjects with muscle contraction/tension-type (MCH) headache and common migraine without aura (CM). Descriptive survey. Chiropractic outpatient research clinic. Forty-seven (47) subjects, aged 18-55 with two categories of benign headache, were studied: MCH (tension-type) n = 19 (6 males, 13 females) and CM (without aura), n = 28 (3 males, 25 females). Subjects were recruited as part of an intervention trial and, thus, form a consecutive sample of patients. The present findings were elicited as part of the initial assessment. No therapeutic intervention is reported. Standardized headache history; plain film and dynamic spinal X rays; motion palpation; and pressure algometry. For CM, the most prevalent headache locations were frontal (81%) and occipital (78%). Neck pain and upper back pain accompanied headache in 90% and 41% of subjects, respectively. For MCH, the most prevalent headache locations were occipital (87%) and frontal (81%). Neck and upper back pain accompanied headache in 100% and 27%, respectively, of all subjects. For the total group, 77% of all subjects and 89% of females exhibited a marked reduction, absence or reversal of the normal cervical lordosis. Ninety-seven percent of all subjects exhibited, on dynamic X-ray studies, at least one significant abnormality of segmental mobility from C1 to C7, while 43% exhibited abnormalities at four or more segments. Segmental motion at C0-C1 was reduced in 90% of subjects in flexion and 70% of subjects in extension. On motion palpation, 84% of CM and MCH subjects were found to have at least two major fixations from C0 to C2. On pressure algometry, 92% of CM and 85% of MCH had at least one verifiable tender point (TP) in the upper cervical region. The most common locations for TPs were mid-cervical (C2-C3), lateral occipital and suboccipital. Both MCH and CM subjects demonstrate high occurrences of: a) occipital and neck pain during headaches; b) tender points in the upper cervical region; c) greatly reduced or absent cervical curve; and d) X-ray evidence of joint dysfunction in the upper and lower cervical spine. These findings support the premise that the neck plays an important, but largely ignored role in the manifestation of adult benign headaches. A case-control study should be conducted to confirm the greater prevalence of cervicogenic dysfunction in headache as compared to nonheadache subjects.


Timolol-pilocarpine fixed-ratio combinations in the treatment of chronic open angle glaucoma. A controlled multicenter study of 48 weeks. Scandinavian Timpilo Study Group. The effect of the fixed combination of 0.5% timolol maleate with 2% pilocarpine hydrochloride given twice a day and 0.5% timolol with 4% pilocarpine given twice a day was tested in a long-term, multicenter study. A total of 360 patients with open angle glaucoma were included; they were defined as those who had intraocular pressure greater than 21 mm Hg while receiving single, topical antiglaucoma therapy. Primarily, the purpose of the study was to investigate the efficacy of 0.5% timolol-2% pilocarpine, and second, to see to what extent an increase in concentration to 0.5% timolol-4% pilocarpine would further lower intraocular pressure in those patients with an intraocular pressure of greater than 21 mm Hg while taking 0.5% timolol-2% pilocarpine. The cohort of 228 patients went through the examinations for a total of 48 weeks. A mean decrease in intraocular pressure from 24.7 +/- 2.8 to 21.0 +/- 3.8 mm Hg was observed. During the trial, approximately 33% of the patients required an increase in concentration to 0.5% timolol-4% pilocarpine after the week 8 examination. At week 12, in those using 0.5% timolol-4% pilocarpine, an additional 2.2 mm Hg lowering of intraocular pressure was observed. Side effects were minor and temporary and did not necessitate withdrawal from the study.


What is the aim of esophageal variceal sclerotherapy--prevention of rebleeding or complete obliteration of veins? Endoscopic variceal sclerotherapy (EVS) is highly effective in arresting active esophageal variceal bleeding. Subsequent repeated EVS sessions significantly reduce recurrence of bleeding; long-term survival is reported as prolonged or unaltered. In contrast, EVS for preventing first variceal bleeding (prophylactic sclerotherapy) is not recommended, even for high-risk patients, because it enhances mortality by significantly increasing the risk of bleeding in the treated group compared with controls. The risk of variceal rebleeding is maximum within 6 weeks of index bleeding; most patients who survive 6 weeks of variceal bleeding (with or without sclerotherapy) behave like patients who have never bled from varices. Thus, EVS continued beyond 6 weeks, to obliterate the veins completely, is akin to prophylactic sclerotherapy. Significant complications of EVS--its cost, discomfort to patient, and loss of doctor and patient time--should be weighed against doubtful benefit of continuing prolonged EVS beyond 6 weeks, just to completely obliterate esophageal veins. The possibility of better long-term survival with sclerotherapy limited to 6 weeks cannot be excluded, because prophylactic sclerotherapy shortens long-term survival.


Signal delivery/real ear measurement system for hearing aid selection and fitting. This report describes a signal delivery/real ear measurement system for application in the hearing aid selection and fitting process. This signal delivery/real ear measurement system provides a means for quantifying a listener's auditory characteristics (e.g., thresholds, loudness discomfort levels) in a manner that is compatible with electroacoustic measures of hearing aid performance. The signal delivery/real ear measurement system consists of a button-type hearing aid receiver coupled to the listener's personal earmold, and a probe tube microphone system to measure the signal level within the occluded ear canal. This signal delivery/real ear measurement system was used to measure detection thresholds and loudness discomfort levels in severe/profoundly hearing-impaired school-age children, with results indicating good test-retest reliability in behavioral responses. The findings presented in this report relate to the intersession electroacoustic variability associated with this instrumentation. In addition, the potential application of this or a similar system for measuring real ear to 2-cc coupler difference values is illustrated and discussed.


Serotonin, histamine and platelets in vascular disease with special reference to peripheral vascular disease. Cardiovascular disease is a major cause of death. There is evidence that this disease is predicted and its progression influenced by various factors (e.g. hyperlipidaemia). In this review, we consider aspects of platelet structure and function which may explain how this cell type contributes to the pathogenesis of vascular disease. The platelet also contains bioamines (serotonin, 5-HT; histamine) which are potent vasoactive substances. Studies involving patients with peripheral vascular disease (PVD) where abnormalities in platelet function (platelet aggregation and platelet shape change) and in bioamine status (vascular, platelet and plasma bioamine concentrations) are reviewed. We also discuss how platelet activation (in vitro) and plasma lipids influence intraplatelet bioamine status. Finally, we report in vitro evidence of the effect of two drugs prescribed to PVD patients: aspirin and naftidrofuryl. Aspirin is an ineffective inhibitor of 5-HT-induced whole blood platelet aggregation whereas naftidrofuryl is effective in the presence or absence of aspirin. By identifying and altering the factors which contribute to the pathogenesis of atherosclerosis we will be better equipped to prevent, reverse or retard this process.


How do peptides interact with lipid membranes and how does this affect their biological activity? 1. A short review is given of the chemical, physical, and pharmacological development of the idea that target cell lipid membranes may catalyze the interaction between regulatory peptides (or other pharmacologic agents) and their cell surface receptors. 2. The message-address and the membrane compartments concepts explain the observed correlations between the three-dimensional structures of peptides induced by a membrane surface and their preference for a certain receptor subtype. 3. Examples are given for opioid peptides (enkephalin, dynorphin, etc.), tachykinin peptides (substance P, neurokinin A, etc.), and melanocortin peptides (ACTH, alpha-MSH, etc.). 4. Relationships between the conformation of substance P induced by membrane association and that of a non-peptide substance P mimetic are discussed. Possible reasons for the difference between agonistic and antagonistic properties in the peptide field are revealed by this case.


Alterations in the levels and lipid composition of plasma lipoproteins (VLDL, LDL and HDL) in Brazilian patients with hepatosplenic schistosomiasis mansoni. 1. The plasma concentrations of low- and high-density lipoproteins (LDL and HDL) were significantly reduced in Brazilian patients with compensated hepatosplenic schistosomiasis mansoni (SM) when compared with healthy individuals, but very low-density lipoprotein (VLDL) levels were unchanged. 2. All three classes of lipoproteins isolated from SM plasma had an increased content of triacylglycerol and unesterified cholesterol and decreased cholesteryl ester and phospholipid. 3. The individual phospholipid composition of patient VLDL, LDL, HDL was also altered; the amount of phosphatidylcholine was increased and that of lysophosphatidylcholine decreased. 4. The saturated and monounsaturated fatty acyl content of cholesteryl esters in patient lipoproteins was also significantly increased, and diunsaturated and polyunsaturated fatty acyl content was decreased. 5. When isolated lipoproteins were examined as negatively stained preparations by electron microscopy, the morphology of SM patient LDL was normal but the HDL fraction was abnormal and showed marked heterogeneity of size with the presence of occasional discoidal particles which resembled "nascent" HDL.


Inhibition of aminopeptidase activity by aromatic and other cyclic compounds. The effect of 2-naphthylamine, p-nitroaniline, o-phenanthroline, sodium deoxycholate and hydrocortisone succinate on the activity of human urine aminopeptidase, rat kidney methionyl and arginyl aminopeptidase, soybean and Enterolobium contortisiliquum seed aminopeptidase was studied using aminoacyl-2-naphthylamide and L-Leu-p-nitroanilide as substrates. Ki values ranged from 10 microM to 2.7 mM. On the basis of Ki and Km values, and catalytic efficiency for each enzyme, it is clear that the aminopeptidases from human urine and from soybean seed should be assayed with both substrates, whereas L-Leu-p-nitroaniline is a more appropriate substrate for the rat kidney aminopeptidases. Sodium deoxycholate is a better inhibitor than hydrocortisone succinate. Non-competitive inhibition was observed in all cases except for E. contortisiliquum seed aminopeptidase.


An improved electrophoretic method for the determination of serum milk protein variants in Gyr-Holstein cows. Milk serum proteins such as alpha-lactalbumin (ALA) and beta-lactoglobulin (BLG) present biochemical polymorphism which is under the control of codominant autosomal alleles. In the present report, we propose modifications of traditional electrophoretic techniques such as increasing the running gel concentration from 5 to 10% and the addition of 5 M urea to the stacking gel, which permitted the detection of two variants (A and B) at the ALA and BLG loci. About 8 microliters of milk serum (6 mg/ml protein) and 10 microliters of total fresh milk were applied. Bovine serum albumin (BSA) and immunolactoglobulins (ILG) could also be discriminated. Total fresh milk was as useful as the purified serum milk proteins for the discrimination of ALA and BLG serum milk protein polymorphism by alkaline vertical slab polyacrylamide gel electrophoresis. However, BSA and ILG ran with caseins, which prevented their characterization in this system.


Difference in the sensitivity of junctional and longitudinal sarcoplasmic reticulum Ca(2+)-ATPase to ADP. The Ca2+ release mechanism that triggers muscle contraction is still not completely understood. We compared Ca2+ accumulation and acetyl phosphate hydrolysis by the Ca(2+)-ATPases present in the longitudinal and junctional membrane of the sarcoplasmic reticulum of rabbit skeletal muscle and found that Ca(2+)-ATPase is more sensitive to ADP inhibition when the enzyme is located on the junctional membrane than when the enzyme is located on the longitudinal membrane (K0.5 = 144 microM for the junctional enzyme vs K0.5 = 415 microM for the longitudinal enzyme). When the enzyme was solubilized in non-ionic detergent (2% v/v Triton X-100) and tested again using 2 mM AcP as substrate, the difference in ADP sensitivity observed with native preparations disappeared. We conclude that the enzyme is regulated differently depending on its localization on the membrane of the sarcoplasmic reticulum.


Effects of testosterone on growth hormone secretion and somatomedin-C generation in prepubertal growth hormone deficient male patients. 1. The role of testosterone (T) in growth was evaluated in 11 prepubertal hypopituitary males during two 15-day periods separated by a 4-week interval, i.e., before (PRE-T period) and during T ester treatment (50 mg every 5 days, 3 im doses-T period). 2. T increased growth hormone (GH) secretion, assessed by 4-h rhythm (mean +/- SEM = 1.90 +/- 0.27 vs 1.77 +/- 0.21 ng/ml; P < 0.05) and after a GHRH stimulus (3.42 +/- 0.54 vs 3.08 +/- 0.43 ng/ml; P < 0.05) as compared to the PRE-T period. 3. T also increased basal somatomedin-C (SM-C) levels (0.20 +/- 0.03 vs 0.15 +/- 0.02 U/ml; P < 0.001) and SM-C generation. After GH was administered in 4 im doses (0.01, 0.02, 0.05 and 0.1 U/kg), SM-C levels were 0.31 +/- 0.08 vs 0.24 +/- 0.07 U/ml, P < 0.001. T did not change incremental (absolute minus basal) SM-C levels (0.15 +/- 0.08 vs 0.12 +/- 0.07 U/ml; P > 0.05). 4. The results suggest that T increased plasma SM-C levels by stimulating residual GH secretion in hypopituitary males.


Abnormal proteins in the cerebrospinal fluid of a patient with Creutzfeldt-Jakob disease following administration of human pituitary growth hormone. A Brazilian case of Creutzfeldt-Jakob disease in a hypopituitary patient who had received cadaver-derived human pituitary growth hormone between 1968 and 1977 is reported. The clinical diagnosis was confirmed during his lifetime by the demonstration of two abnormal 30-kDa proteins in the cerebrospinal fluid by two-dimensional gel electrophoresis. These proteins, characteristic of Creutzfeldt-Jakob disease, present isoelectric points of 5.1 and 5.2. Furthermore, both proteins migrate as doublets, each one displaying a molecular weight variant of about 29-kDa. This is one of 16 cases of the disease associated to therapy with cadaver-derived human growth hormone and one of the few examples among such cases of confirmation of the clinical diagnosis by biochemical characterization of abnormal proteins in the cerebrospinal fluid.


Effect of purified components of Ascaris suum extracts on IgE antibody response in guinea pigs. The effect of components P530 and P29, isolated from Ascaris suum adult worm extract (ASC), on the heterologous IgE antibody response was studied in guinea pigs. Groups of 7 guinea pigs were immunized ip with 50 micrograms of ovalbumin (OA) alone or mixed with 200 micrograms of each component precipitated in an alum gel. The primary and secondary IgE antibody response was evaluated by passive cutaneous anaphylaxis reaction (PCA). Immunization of guinea pigs with P29 plus ovalbumin (OA) resulted in a significant increase in the level of serum IgE anti-OA antibodies, especially in the secondary response (almost 8-fold higher when compared with control group). This potentiated response was not observed when the animals received OA plus P530 or the crude extract. Indeed, the P530 component, as well as the crude extract, induced a depression of the anti-OA IgE antibody response (2-3 fold decrease when compared with OA-immunized animals). It was also shown that P29, but not P530 or ASC, was capable of eliciting a strong anti-ASC IgE antibody response. These results demonstrate that in guinea pigs these two Ascaris suum components have antagonistic biological effects, one inducing potentiation and the other suppression of the heterologous IgE antibody response.


Mathematical dose-response model for sodium cotransport with glucose and glutamine in the rabbit ileal mucosa. Hyperbolic and allosteric mathematical equations are used extensively to describe cumulative dose-response curves in pharmacology. In 1954, using an axiom described by Clark, Ariëns suggested the following equation for cumulative dose-response curves: Em/E-1 = Kd/[D], where Em = maximum effect, E = effect, Kd = dissociation constant of the drug-receptor complex and [D] = drug concentration. To evaluate the validity of the modified Ariëns equation on the dose-response curves with a Hill coefficient statistically different from one, we used sodium intestinal cotransport (SIC) with glutamine (Gln) and glucose (Glu). Increasing doses (10 microM to 0.1 M) of Gln and Glu were added to the mucosal side of the isolated rabbit ileum mucosal membrane mounted on Ussing chambers. Sodium cotransport was recorded by measuring short-circuit current (Isc), potential difference (PD) and tissue resistance (TR) at 20-min intervals for 2 h. The Em and ED50 for SIC with glucose were 25% and 150 microM, respectively, using the method proposed by Lineweaver-Burk. The same method could not be used to estimate the Em and ED50 in SIC with glutamine, suggesting a specific limitation of the Gln cotransport kinetic curve in the equation. Therefore, we modified the Ariëns equation to Em/E-1 = (Kd/[D])p. With p = Log (S), Log (Kd) = -pD2 and Log[D] = -pDx, we obtain E = Em/(1 + S(pD x -pD2)), where S is a hyperbolic constant.(ABSTRACT TRUNCATED AT 250 WORDS)


The N-terminal amino acid sequence is essential for foot-and-mouth disease virus replicase activity. 1. Foot-and-mouth disease virus replicase was expressed by fusing its cDNA to the OmpA signal peptide coding sequence present in the pIN-III ompA series vectors. 2. Two constructions were developed to express either a full-length or truncated enzyme lacking the 20 amino acids at the N-terminal end. Bacterial extracts expressing the recombinant proteins were submitted to SDS-PAGE and the presence of the replicase was revealed by immunoblotting. The truncated form exhibited a higher mobility and the relative positions of the proteins show that the signal peptide was removed. 3. The biological activity of these two molecules was tested using a poly(A)-dependent oligo(U)-primed poly(U)-polymerase assay. The full-length replicase is active. The aminoterminal truncated enzyme had 0.02% activity of the intact one. 4. This result indicates the importance of the twenty N-terminal amino acids for the activity of FMDV RNA-dependent RNA polymerase.


Use of plasmid profiles to differentiate strains within specific serotypes of classical enteropathogenic Escherichia coli. 1. The usefulness of plasmid profile analysis to differentiate strains of enteropathogenic Escherichia coli (EPEC) was evaluated by studying 123 strains of the most prevalent serotypes causing infant diarrhea in the city of São Paulo, Brazil, i.e., O111ab:H-, O111ab:H2 and O119:H6. 2. No common profiles were found among strains of distinct serotypes. However, within each serotype, most of the strains were grouped within a few major profiles. More than 68% of the strains of serotypes O111ab:H- and O111ab:H2 were included in 6 and 9 major profiles, respectively. In serotype O119:H6, about 48% of the strains were included in 3 major profiles. 3. This analysis suggests that only a few EPEC clones are causing infant diarrhea in São Paulo and revealed that the distribution of serotypes O111ab:H- and O111ab:H2 during the one-year study was at least partly determined by small outbreaks of the most common profiles. 4. We conclude that plasmid profile analysis is very useful to differentiate strains within specific EPEC serotypes.


The relative importance of familial, reproductive and environmental factors in biliary atresia: etiological implications and effect on patient survival. 1. The clinical records of 237 patients with extrahepatic biliary atresia (EHBA) attending King's College Hospital, London, between March 1973 and October 1985 were analyzed in terms of familial and reproductive factors, as well as of their possible effect on patient survival. 2. The male:female ratio was 0.93, and the ages of the patients' mothers and fathers were significantly higher than would have been expected from sibship data. Similarly, the number of first-born children having EHBA was less than expected. 3. In a subsample of 189 mothers, the obstetrical histories of women who had children with associated EHBA (20% of the total) showed: 1) a higher frequency of illness before and during pregnancy; 2) a higher level of pharmaceutical drug consumption during pregnancy, and 3) more fetal losses, especially in the gestation immediately preceding the patient's birth. 4. Age at death (average 15.1 +/- 13.2 months) and survival rates depend critically on surgical intervention and were not related to the presence or absence of extrahepatic malformations or to the type of atresia. 5. The present observations, taken together with those of others, indicate that problems in the reproductive process or exposure to noxious environmental agents may be etiological factors in associated EHBA.


Indirect immunofluorescence determination of the frequency of anti-H. pylori antibodies in Brazilian blood donors. 1. The accuracy of an indirect immunofluorescence (IIF) test for the serodiagnosis of Helicobacter pylori infection was evaluated in adult patients and compared with culture, preformed urease test and smears stained with carbolfuchsin. 2. We studied 80 consecutive patients submitted to gastroduodenoscopy (17 of whom were found to have duodenal ulcer) plus 57 patients with duodenal ulcer. 3. H. pylori was identified by microbiological methods in 65 of the 80 consecutive patients and in all 57 patients with duodenal ulcer. 4. Among the 74 patients with duodenal ulcer, 71 presented antibody titers > or = 1:20 and 46 of the 48 H. pylori-positive patients without duodenal ulcer presented antibody titers > or = 1:20. 5. Thirteen of the 15 H. pylori-negative patients presented antibody titers < or = 1:10. 6. The sensitivity, specificity and positive predictive value of the IIF test were 95.9%, 88.8% and 98.4%, respectively. 7. The seroprevalence of H. pylori in 380 asymptomatic Brazilian blood donors was also studied by the IIF test. The presence of IgG antibodies against H. pylori was observed in 62.1% of the individuals. The prevalence of H. pylori infection increased with age and no difference was observed between males (60.3%) and females (66.6%).


Primary isolation of a Brazilian strain of hepatitis A virus (HAF-203) and growth in a primate cell line (FRhK-4). 1. Although hepatitis A is endemic in Brazil, this is the first report describing the isolation of a Brazilian strain of hepatitis A virus (HAV). 2. Fecal specimens obtained from patients in the acute phase of hepatitis A were inoculated into fetal Rhesus kidney cell cultures (FRhK-4). Only one inoculum, denoted HAF-203, could be propagated serially. Both cell lysates and tissue culture fluids of infected cells were used as inocula and evaluated for viral antigen and RNA content by enzyme immunoassay and cDNA-RNA hybridization, respectively. 3. Cell lysates gave better yields when used as viral inocula. After three passages, viral RNA and antigen were detected in cell lysates 4 and 14 days post-infection, respectively. Using tissue culture fluid as inoculum, the incubation period was decreased from 49 to 7 days after 4 serial passages, reflecting the adaptation of HAF-203 to growth in FRhK-4 cells. 4. FRhK-4 cells can now be used for HAV antigen production for diagnostic assays and molecular characterization.


The effect of crotoxin on the release of acetylcholine and lactate dehydrogenase from rat brain cortical slices. 1. We have studied the effects of crotoxin, the neurotoxin of the South American rattlesnake Crotalus durissus terrificus, on the release of acetylcholine and lactate dehydrogenase from rat brain cortical slices. 2. Crotoxin enhances the release of [3H]-acetylcholine from cortical slices (control values 92.8 +/- 5.9 and 150.3 +/- 11.7 DPM/mg and crotoxin values 199.1 +/- 7.0 and 336.0 +/- 26.0 DPM/mg, at 60 and 120 min incubation, respectively) in parallel with the release of lactate dehydrogenase (control values 50.4 +/- 16.8 and 80.3 +/- 19.5 U/mg and crotoxin values 162.5 +/- 39.1 and 355.7 +/- 38.2 U/mg, at 60 and 120 min incubation, respectively). Both effects are markedly reduced when substituting Sr2+ for Ca2+ in the incubation medium. 3. It is concluded that the phospholipase activity of crotoxin is responsible for the observed effects.


Anxiogenic action of acute but not repeated cocaine administration in handling-habituated mice in the plus-maze test. The present study was designed to determine the putative anxiogenic effects of cocaine on the performance of mice in the elevated plus-maze test of anxiety. Male Swiss mice, weighing about 30 g, treated repeatedly with saline for 8 days and challenged with a single injection of cocaine 10 mg/kg, ip, 1 ml/100 g, on day 10, showed a clear reduction in the relative number (%) of entries and time spent on the open arms of the maze (30.5 +/- 4.5 and 29.4 +/- 5.0 vs 47.1 +/- 6.2 and 46.7 +/- 9.4, for the controls, respectively, N = 7-8 animals). These data suggest an anxiogenic action of cocaine. However, acute treatment with cocaine (5-20 mg/kg, ip) of naive mice and repeated administration of cocaine (10 mg/kg) for 8 days failed to alter the indices of anxiety in the test. These findings suggest that the emotional changes induced by the handling procedures for 8 days may be related to the occurrence of anxiogenic responses to cocaine by mice.


Electromechanical effects of urea on the isolated rat heart. 1. Langendorff-perfused isolated hearts and left ventricle papillary muscles from 3-month-old albino rats of both sexes were studied before and after a 30-min treatment with 17 mM urea added to the medium, a concentration equivalent to a plasma level of 100 mg/dl. To determine whether the effects observed after the addition of urea were produced by an increase in tonicity, the study was repeated using 17 mM sucrose. 2. Mechanical studies on the papillary muscles showed that isometric force development and its first time derivative decreased after washing out urea from the bath (F = 9.73 +/- 1.02 g/mm2 to 7.47 +/- 0.72 and dF/dt = 66.8 +/- 6.43 to 56.7 +/- 4.60 g (mm2)-1 s-1, respectively; P < 0.05). Inotropic responses to isoproterenol and increased extracellular calcium after urea treatment reached values similar to those obtained before urea treatment. Thus, the effect of isoproterenol and calcium was stronger than that obtained before urea treatment. 3. In Langendorff-perfused hearts, the spontaneous heart rate did not change after urea or sucrose treatment. Urea promoted a decrease in the left ventricle isovolumic systolic pressure (39.7 +/- 4.05 to 26.1 +/- 2.69 mmHg, P < 0.05) and a reduction of total QRS amplitude. 4. In both papillary muscles and isovolumic perfused hearts, contractile responses resulting from changes in extracellular sodium concentration were reduced after urea treatment. The increased osmolarity due to sucrose did not produce any changes in electromechanical activities. 5. Although 17 mM, which reduces isometric force and isovolumic pressure development and modifies the ECG, is well below the concentration required to modify protein conformation in vitro, the present results suggest that its action could be explained by an effect at the sarcolemmal level.


Technical problems related to the analysis of the effects of inorganic phosphate on cardiac muscle. 1. We describe how potential artifacts (due to solution composition, buffering capacity of the bathing medium, size of the skinned fiber preparation, permeability of the sarcoplasmic reticulum (SR) vesicles, and proper Kd for Ca2+ of the fluorescent indicator used to measure Ca2+ transport can be avoided in order to determine the effects of inorganic phosphate (Pi, or any other ion) on maximum Ca2+ activated force (Fmax) and Ca2+ sensitivity of skinned cardiac muscle fibers, and Ca(2+)-ATPase activity and uptake properties of isolated cardiac SR-enriched vesicles. 2. To maintain the ionic strength of the bathing medium constant when adding Pi, other ions must be removed. We found that because some salts have a depressant effect on Fmax independent of increased ionic strength (e.g. KCl) while others do not (e.g. Na-acetate), the salt used to adjust ionic strength influences the measured depressant effect of Pi on Fmax. 3. The sensitivity to Ca2+ of the contractile apparatus depends on the sum of the [Na+] and [K+] in the bathing medium. However, we found that the effect of Pi on Ca2+ sensitivity was not significantly influenced by the small changes in the sum of [Na+] and [K+] that were associated with the addition of Pi. 4. The skinned fiber preparations were approximately cylindrical bundles with diameters ranging between 100 and 250 microns. We found that the effect of Pi on Fmax was not influenced by diffusion limitations over this range of bundle diameters. 5. The pH buffering capacity of the bathing solution affects Fmax at pH 6.6. We found that the buffering effect of Pi can influence the mechanical response of skinned fibers independent of a direct effect of Pi on the contractile apparatus when the buffering capacity of the control solution is low. 6. When the Ca(2+)-ATPase of isolated SR vesicles is activated by Ca2+ and MgATP, the vesicles accumulate Ca2+. Unless the vesicles are permeabilized with a Ca2+ ionophore (ionomycin) and the pH adequately buffered, maximum ATPase activity will be underestimated, the broadness of the curve relating Ca2+ to Ca(2+)-ATPase rate overestimated, and the sensitivity to Pi overestimated. 7. The ionic milieu of isolated SR vesicles changes the apparatus dissociation constant (Kd) of Ca(2+)-Fura-2, a fluorescent dye used to quantify SR Ca2+ transport rates. In order to accurately measure the inhibitory effect of Pi on Ca2+ uptake, the influence of Pi on the Kd of fura-2 for Ca2+ must be taken into account.


Theta waves and behavioral manifestations of alertness and dreaming activity in the rat. Electro-oscillographic recordings show that when rats wake up, behavioral manifestations such as head dorsal extension and snout and eye movements (which occur in this sequence), occasionally followed by brief ear and limb twitches, are preceded by and are simultaneous with theta waves (around 6.5 Hz in frequency) in neocortical areas 3 and 17 and in the hippocampus. Heart rate increases when the animal passes from synchronized sleep to relaxed wakefulness to exploratory behavior. During desynchronized sleep theta waves, similar to those found during alertness, usually precede and are simultaneous with rostrum and eye movements and brief ear and limb twitches. Heart rate also increases during dreaming activity. A high degree of coherence (near 1.0) was found among theta bursts in different hippocampal areas in both highly attentive alertness and dreaming. It is suggested that theta waves are related to attention as part of some kinds of behavior and of the process of dream shaping and expression, both characterized by consciousness.


Theta waves in the Papez circuit during alertness and dreaming in the rat. An electro-oscillographic study of the hippocampus, thalamic anterior nuclei and cingulate and pericingulate cortical areas was performed in the rat during the wakefulness-sleep cycle. High voltage (usually from 100 to 200 microV) theta waves, oscillating at regular frequencies from 6 to 10 Hz, were found to occur simultaneously in all of these components of the Papez circuit during attentive behavior and dreaming.


In vitro activities of a recombinant foot-and-mouth disease virus replicase expressed in Escherichia coli. 1. The replicase gene of foot-and-mouth disease virus (FMDV) was expressed in Escherichia coli under the control of a tac promoter. The recombinant enzyme was purified by inclusion body precipitation, elution, and poly(U) Sepharose chromatography. 2. The enzyme exhibits poly(A)-dependent oligo(U)-primed poly(U) polymerase activity. The specific activity of the purified replicase is 1.3 x 10(5). The recombinant replicase synthesizes RNA using FMDV RNA as template, as well as heterologous RNAs, such as globin RNA and synthetic RNAs, polyadenylated or not. In all polymerization reactions, RNA products twice the size of the template are formed, both in the presence and absence of an oligo(U) primer. The enzyme is also capable of incorporating [alpha 32P]UTP in all RNAs tested except the viral template. This activity does not seem to be related to the primer independent polymerization activity. 3. The products from polymerization reactions were characterized by hybridization. In the absence of primer they consist of the template and a complementary strand covalently attached, while in the presence of primer they consist of two complementary strands synthesized de novo. 4. We propose mechanisms of RNA synthesis by the recombinant FMDV replicase in the absence and presence of primer. These mechanisms are discussed in terms of models for in vitro RNA synthesis of other picornaviruses.


Cloning of a B10 DNA puff sequence developmentally amplified and expressed in the salivary gland of Bradysia hygida. A recombinant clone carrying a 2-kb fragment was isolated from a mini-library of the B10 DNA puff of Bradysia hygida. This fragment was amplified in the salivary gland during the period of DNA puff formation. Amplification started when DNA puff anlage was formed and continued to increase, reaching a maximum of about 10-fold 28 h later. Northern blot hybridization experiments showed that this 2-kb fragment was complementary to two RNA species of about 1.3 kb and 1.1 kb, which are developmentally regulated in the salivary gland. Maximum amounts of these messages were present when the B10 puff is fully expanded.


Calcium transport by HeLa cell mitochondria. 1. We studied mitochondria of digitonin-permeabilized HeLa cells, since digitonin (60 micrograms/10(6) cells) increases plasma membrane Ca2+ permeability, in order to avoid problems such as low mitochondrial yield and the possibility of obtaining damaged or uncoupled mitochondria from tumor cells. 2. Addition of Ca2+ to digitonin-permeabilized HeLa cells gave rise to a cycle of respiratory stimulation. Ca2+ uptake was almost totally inhibited by antimycin A (6.0 micrograms/ml). 3. Ca2+ release occurred upon addition of carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP) to digitonin-permeabilized HeLa cells, under steady-state conditions. An antimycin A- and FCCP-insensitive Ca2+ uptake was also detected in this preparation when ATP was added, which reflects Ca2+ capture by other organelles. 4. The characteristics of the mitochondrial Ca2+ transport system in HeLa cells are similar to those of other previously studied tumor cells. Mitochondria from HeLa cells are resistant to the deleterious effects of massive Ca2+ loads.


A rapid test for the detection of a B-cell marker (D8/17) in rheumatic fever patients. A new, rapid blood test for the detection of B-cell markers in patients with rheumatic fever (RF) was applied to 10 patients with RF, 26 of their family members and 14 normal subjects. Three-hundred microliters of blood was obtained by finger-pricking from each subject and submitted to the immunofluorescence technique using three different antibodies, i.e., D8/17 monoclonal ascites antibody, FIRC-conjugated goat anti-mouse IgM (mu chain specific) and phycoerythrin-conjugated murine anti-HLA-DR antibody. D8/17 lymphocytes (green stain) and B-lymphocytes (red stain) were counted using a double-filter microscope. The percentage of positive cells was 20-64% (average 38.5%) in patients, siblings, fathers and mothers, 4.5% in 12 normal controls and 27.5% in the other 2 controls with a family history of RF. In 90% of the families studied, the patients had the highest percentage of positive cells. We conclude that this rapid test can be used as an aid for RF diagnosis and possibly to detect siblings who are at risk to develop the disease.


Effect of an essential fatty acid deficient diet on experimental infection with Trypanosoma cruzi in germfree and conventional mice. 1. To investigate the possible role of essential fatty acid deficiency in host cell/parasite interaction, weanling germfree (GF) and conventional (CV) CFW mice maintained on an essential fatty acid-deficient (-) or a control (+) diet for 110 days were infected with Trypanosoma cruzi. 2. Blood parasitemia indicated that the disease was milder in the animals maintained on the essential fatty acid-deficient diet than in the GF and CV controls (maximum parasitemia: GF+ 33,300, GF-26,200, CV+ 17,100 and CV- 6,400 trypomastigotes/ml blood). 3. Survival 30 days after infection was 12% for GF+, 28% for GF-, 37% for CV+ and 65% for CV- mice. 4. Linoleic and arachidonic acid levels were significantly lower in animals kept on the essential fatty acid-deficient diet (GF-: 28.0 +/- 9.3, 23.4 +/- 8.6; CV-: 37.6 +/- 5.8, 19.9 +/- 3.6) than in controls (GF+: 164.4 +/- 48.8, 162.6 +/- 45.8; CV+: 147.1 +/- 26.5, 107.5 +/- 23.6) confirming the deficiency. 5. Before the infection, succinic dehydrogenase levels were higher in liver of all CV mice (4.52 micrograms phosphate/mg fresh tissue) than in GF mice (0.84 micrograms phosphate/mg fresh tissue), whereas the opposite was true for 5'-nucleotidase levels in brain and liver, respectively (GF: 2.84 and 3.18 micrograms phosphate/mg fresh tissue; CV: 1.25 and 1.54 micrograms phosphate/mg fresh tissue). 6. The disease was milder in deficient than in control animals in both the GF and CV groups on the basis of parasitemia and survival, indicating that fatty acid-deficient mice are partially protected against Chagas' disease. The mechanism underlying this phenomenon requires further investigation.


Gangliosides inhibit serological reactions for the detection of cholera and heat-labile enterotoxins of Escherichia coli. GM1 ganglioside has been identified as the receptor for cholera toxin (CT) and heat-labile (LT) enterotoxin of Escherichia coli in many cell types. Using the radial immune hemolysis (RIH) and indirect hemagglutination (IH) tests described for the detection of these enterotoxins, a study was conducted on the 100% inhibition of these reactions by pre-incubating these enterotoxins with GM1, GD1a and GT1 gangliosides. GM1 was found to be much more efficient than the other two. With respect to the RIH test, GT1 was more efficient than GD1a as an inhibitor of enterotoxin binding. Similar results were obtained with the IH test. These data also showed that sheep red blood cells provide a good model system for the study of receptors for CT, LT and probably other enterotoxins which bind to gangliosides.


Inhibition of enteropathogenic Escherichia coli adherence to HeLa cells by immune rabbit sera. We have studied the effect of immune rabbit sera on the localized (LA) and diffuse (DA) adherence to HeLa cells of 10 enteropathogenic Escherichia coli (EPEC) strains belonging to serogroups O55, O86, O111, O119, and O142. Anti-La1 serum, obtained by rabbit immunization with an E. coli strain harboring a cloned DNA fragment from an EPEC LA plasmid, strongly inhibited the adherence of all serogroups but one (O142). Similar results were obtained with anti-LA2 serum, which is anti-O111 serum absorbed with a non-adherent O111:H- EPEC strain. In contrast, non-absorbed anti-O55 and anti-O111 sera showed an inhibitory effect mainly on the adherence of homologous strains. Except for one experiment diffuse adherence was not inhibited by any antiserum used. The inhibitory effect of immune sera on localized adherence does not seem to be correlated with plasmid curing since adherence plasmid pMS49 proved to be stable after treatment with anti-O55 and anti-O111 sera. The cross-inhibition of adherence by anti-LA sera suggests that localized adherence-related adhesins of the O55, O86, O111, and O119 strains share similar antigens.


Influence of age on the induction of oral tolerance in mice and its adoptive transfer by spleen cells. 1. Seven-week-old B6D2F1 mice were highly susceptible to the induction of oral tolerance to ovalbumin (Ova), whereas 70-week-old mice were totally refractory. 2. Immune responsiveness (secondary antibody formation) to intraperitoneal immunization to Ova was the same in 7-week- or 70-week-old B6D2F1 mice. 3. In B6D2F1 mice, the adoptive transfer of spleen cells from old donors into young recipients hindered, and, reciprocally, transfer of spleen cells from young donors into old recipients facilitated the induction of oral tolerance. 4. In BALB/c mice, which are refractory to oral tolerance to Ova, the adoptive transfer of spleen cells from neonate or young donors into old recipients failed to modify the lack of susceptibility to the induction of oral tolerance.


Differences in macrophage stimulation and leukocyte accumulation in response to intraperitoneal administration of glucose/mannose-binding plant lectins. Peritoneal macrophage stimulation (rapid spreading on glass surface and hydrogen peroxide production) and inflammatory reaction (leukocyte accumulation) obtained in C3H/HeJ mice at 8 weeks of age, after a single ip injection of 10 micrograms concanavalin A (Con A), a lectin extracted from Canavalia ensiformis, were compared with those obtained with two other glucose/mannose-binding lectins extracted from Canavalia brasiliensis (Con Br) and Dioclea grandiflora (DGL). All lectins enhanced macrophage spreading 3- to 4-fold at 24-72 h compared to control. Stimulation of hydrogen peroxide release by Con A, Con Br and DGL lasted 1, 2 and 3 days, respectively. Leukocyte cell influx at 24-72 h after lectin injection consisted mainly of mononuclear cells. Con A induced a moderate increase in the total number of peritoneal cells, whereas administration of Con Br or DGL increased the number of peritoneal cells 2- to 3-fold. The results indicate that DGL and Con Br have more pronounced effects on macrophage stimulation and inflammatory reactions than Con A.


Effect of age on antinociceptive effects of elevated plus-maze exposure. It has been suggested that anxiety may be a critical factor in certain forms of non-opioid environmental analgesia. Furthermore, age has been reported to increase the anxiety levels in rats as measured in the elevated plus-maze. In the present investigation 10 young (3 months), 10 middle-aged (14-16 months) and 10 old (28-30 months) male Wistar rats were tested by the tail withdrawal assay of nociception before (baseline), and at 0 (T1) and 10 (T2) min after a 5-min exposure to the elevated plus-maze apparatus. Only old rats presented an increase in tail withdrawal latencies after elevated plus-maze exposure, even though this effect was statistically significant only immediately after exposure to the apparatus (baseline = 2.5 +/- 0.3 s; T1 = 3.8 +/- 0.3 s; T2 = 3.3 +/- 0.4 s). The results indicate that exposure to the elevated plus-maze induces a rapidly reversed and age-dependent antinociception in rats. They are also consistent with the proposed greater sensitivity of old rats to anxiogenic effects of the plus-maze.


Anxiety-induced antinociception in the mouse. It has been suggested that exposure to the elevated plus-maze (EPM) apparatus induces antinociceptive effects in mice as measured by the tail-flick assay, which are not blocked by the opiate antagonist naltrexone. The present study performed on 3-month old male EPM-M1 albino mice (12-14 animals per group) was designed to assess a) if exposure limited to the open or to the enclosed arm of the EPM would alter this effect; b) whether or not pharmacologically induced anxiety (1.0 mg/kg pentylenetetrazole, PTZ) would also reduce nociception; c) if exposure to the EPM would alter visceral pain, as measured by the abdominal contortion test. The simultaneous exposure to both the open and enclosed arms of the EPM, but not the exposure limited to each type of arm, led to statistically significant increases in tail withdrawal latencies (TWL). Indeed, 10 min after exposure to both arms, TWL values (mean +/- SEM) were 10.31 +/- 0.87 s as compared to a baseline value of 5.46 +/- 0.53 s. The acute administration of PTZ significantly increased TWL. Conversely, EPM-induced antinociception was not detected by the abdominal contortion test. These results confirm the existence of EPM-induced antinociceptive effects demonstrated by others and show that they may be influenced by multiple determinants.


Correlations between morningness-eveningness character, sleep habits and temperature rhythm in adolescents. The correlations between morningness-eveningness character (Horne & Ostberg score), sleep habits and temperature acrophase of 62 adolescents (mean age 13 years and 6 months) were investigated on three occasions at 6-month intervals. Horne & Ostberg scores ranged from 29 to 74 with a similar distribution on the three occasions. Temperature acrophase mode occurred around 15:00 p.m. Earlier sleep onset time and wake-up time and earlier temperature acrophase were associated with morningness. The more evening type the adolescent was, the shorter was the sleep length on schools days and the longer was the sleep length on non-school days. Correlations between temperature acrophase and morningness-eveningness character were found in a subgroup of adolescents and require further confirmation. The correlations between morningness-eveningness and sleep habits were present on the three occasions investigated, indicating that these are stable correlations and that morningness-eveningness questionnaires can provide suitable information for studies of this age group.


Modulation of food and water intake by catecholamines injected into the lateral ventricle of the pigeon brain. The effects of noradrenaline, adrenaline and dopamine on food and water intake were investigated in satiated (food ad libitum) adult pigeons weighing 320-360 g (N = 10 per group). Catecholamines (80 nmol in 1 microliter of saline) were injected into the right lateral ventricle. A significant increase in food consumption during the first hour was induced only by noradrenaline (8.1 +/- 1.8 g) and adrenaline (8.0 +/- 2.7 g) vs 1.9 +/- 0.6 g for the saline control. Noradrenaline also caused a significant drop during the second hour (1.7 +/- 0.6 g vs 3.5 +/- 0.7 g for the saline control). Dopamine decreased food intake at 2 h (1.4 +/- 0.3 g vs 3.5 +/- 0.7 g for the saline control) and 3 h (1.4 +/- 0.4 g vs 3.2 +/- 0.7 g for the saline control) post-injection, with a subsequent elevation in the feeding response during the 4th h (4.9 +/- 0.9 g vs 2.6 +/- 0.7 g for the saline control). The total food ingestion for the 6-h period was similar for the control and experimental groups. Water consumption was not significantly affected by any catecholamine during any specific period but total ingestion was increased by noradrenaline (44.7 +/- 3.0 ml vs 29.6 +/- 5.1 ml for the saline control). These results suggest that catecholamines are involved in the neural control of food, but not water, intake in pigeons.


Evidence for a developmentally regulated neurotrophic activity from chick retina affecting the survival of retinal ganglion cells from newborn rats. The effect of conditioned medium from aggregates of chick embryo retinal cells was tested on the in vitro survival of retinal ganglion cells from newborn rats. Ganglion cells were identified by the detection of retrogradely transported horseradish peroxidase injected bilaterally into the superior colliculus. Culture medium conditioned with chick embryo retinae was tested on monolayers of rat retinal cells with plating densities ranging from 0.5 to 4 x 10(5) cells/cm2. In all cases the conditioned medium significantly increased the survival of ganglion cells after 2 days in culture. Conditioned media from embryonic days 8 to 16 (E8 to E16) presented neurotrophic activity, with the greatest effect occurring at E10-E12. The conditioned medium had no effect on the adhesion of rat retinal cells. The data suggest that chick retinal cells produces soluble trophic factors which can influence the survival of rat retinal ganglion cells in vitro. Furthermore, the release of this neurotrophic activity by chick retina seems to be developmentally regulated.


Further studies on the hypoxia produced by canatoxin in rats. Canatoxin, a convulsant neurotoxin from the seeds of Canavalia ensiformis, induces lipoxygenase-dependent hypoxia in rats which is blocked by hexamethonium. The purpose of the present study was to examine the relationship between canatoxin-induced hypoxia and bronchoconstriction. Since several effects of the toxin are very similar to those described for morphine and opioid-like peptides, the effects of opioid antagonists were also investigated. Pretreatment of male, adult Wistar rats (200-250 g) with cyproheptadine (80 micrograms/kg, ip, N = 6) and isoproterenol (100 micrograms/kg, ip, N = 6) partially blocked (% variation of pO2: CNTX alone: -26.67 +/- 2.56, N = 6; with cyproheptadine: -16.15 +/- 2.97*, N = 6; with isoproterenol: -17.73 +/- 1.93*, N = 6; *P < 0.05 as compared to CNTX alone) the hypoxia but no effect was observed with diphenhydramine (2 mg/kg, ip, N = 6) or atropine (2 mg/kg, ip, N = 6). The hypoxemic effect of canatoxin (100 micrograms/kg, i.v., 20 min, N = 6) was also almost completely blocked with either naloxone (1 mg/kg, sc, N = 6) or naltrexone (5 mg/kg, sc, N = 6). The results presented here provide evidence suggesting that both opioid peptides and bronchoconstriction seem to play a role in the hypoxia caused by canatoxin.


Evidence for the involvement of nitric oxide in the electrically induced relaxations of human lower esophageal sphincter and distal pylorus. The aim of the present study was to investigate in vitro the effect of NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide (NO) synthase inhibitor, on neurogenic relaxation of human lower esophageal sphincter (LES) and distal pylorus (DP) circular muscle strips induced by electrical field stimulation (EFS). Muscle strips obtained from 5 patients who underwent total gastrectomy were suspended in 10-ml organ baths containing Krebs solution for recording isometric tension. L-NAME (30 microM) reduced the amplitude of the EFS-induced relaxation by 85 +/- 9% (N = 3) in the LES and by 52 +/- 16% (N = 3) in the DP but did not affect sodium nitroprusside-induced relaxation. L-Arginine (300 microM) partially reversed the L-NAME inhibition in the LES and totally in the DP. These findings suggest a role for L-arginine-derived NO in the nerve-mediated NANC relaxation of the human LES and DP.


Lesions of the lateral hypothalamus impair the pressor response to clonidine injected into the medial septal area of conscious rats. The central injection of clonidine (an alpha 2-adrenoceptor agonist) in conscious normotensive rats produces hypertensive responses and bradycardia. The present study was performed to investigate the effect of electrolytic lesions of the lateral hypothalamus (LH) on the pressor and bradycardic responses induced by clonidine injected into the medial septal area (MSA) in conscious and unrestrained rats. Male Holtzman rats weighing 250-300 g were used. Mean arterial pressure and heart rate were recorded in sham- or bilateral LH-lesioned rats with a cerebral stainless steel cannula implanted into the MSA. The injection of clonidine (40 nmol/microliter) into the MSA of sham rats (N = 8) produced a pressor response (36 +/- 7 mmHg, P < 0.05) and bradycardia (-70 +/- 13 bpm, P < 0.05) compared to saline. Fourteen days after LH-lesion (N = 9) the pressor response was reduced (9 +/- 10 mmHg, P < 0.05) but no change was observed in the bradycardia (-107 +/- 24 bpm). These results show that LH is an important area involved in the pressor response to clonidine injected into the MSA of rats.


Effects of mercury on the mechanical and electrical activity of the Langendorff-perfused rat heart. The effects of increasing concentrations of mercury (Hg2+) chloride (0.5, 1, 2 and 10 microM) on the myocardial electromechanical activity were studied on 10 Langendorff-perfused rat hearts. Hg2+ decreased the development of isovolumic systolic pressure from 20.3 +/- 2.13 mmHg under control conditions to 6.25 +/- 1.32 mmHg at 10 microM HgCl2 (P < 0.01) (diastolic pressure = 0 mmHg). The atrial and ventricular rates also decreased at 0.5 microM, 1 microM and 2 microM HgCl2 when compared to the Hg(2+)-free solution (from 201 +/- 4 to 126 +/- 15 bpm). However, at 10 microM Hg2+ the atrial rate increased (155 +/- 19 bpm) whereas the ventricular rate did not change significantly (119 +/- 13 bpm). A delay in atrioventricular conduction occurred at 0.5 microM Hg2+ (64 +/- 4 ms in the Hg(2+)-free solution to 91 +/- 14 ms in the presence of 0.5 microM Hg2+, P < 0.05) with no further changes at higher Hg2+ concentrations. The QRS-T duration also increased as a function of increasing Hg2+ concentrations (58 +/- 5.5 ms in the Hg(2+)-free solution to 123 +/- 15 ms in the presence of 10 microM Hg2+, P < 0.01). Qualitative changes of ECG such as extrasystoles, atrial or ventricular arrhythmias and A-V blocks were also observed. The inhibitory action of Hg2+ on ATP hydrolysis and on Ca2+ and Na(+)-K+ ATPases suggested to occur in other tissues could be the mechanism responsible for the observations reported here.


Propranolol reduces myocardial hypertrophy in the right cardiac chambers after infarction in rats. The effects of chronic propranolol (Prop) therapy on the postinfarction myocardial hypertrophy of infarcted rats were studied by histological techniques. Male albino rats were submitted to left coronary artery ligation to produce infarction or to sham surgery (Con, N = 6). Infarcted rats (Inf) were divided into 2 groups receiving Prop (2.5 mg/kg, twice a day, N = 6) or saline (N = 6) for one month, respectively. Myocyte diameters were measured in longitudinally oriented sections in the four heart chambers (60 cells/chamber). Inf produced a significant increase in mean diameter of myocytes from the right atrium and ventricle and from the left atrium. In the right ventricle, myocyte diameter increased from 8.9 +/- 0.5 microns in the Con group to 12.5 +/- 0.6 microns in the Inf group (P < 0.05). Under Prop, myocyte diameter was reduced (P < 0.05) to 9.8 +/- 0.9 microns. Similar values were observed in the right atrium. In the left atrium, Prop produced only a partial reversion of the postinfarction hypertrophy. In the left ventricle, myocyte diameter was not significantly changed after Inf or Prop therapy. These data show that beta blockers reduce the myocardial hypertrophy in the right heart chambers after experimental infarcts in rats. This effect can be secondary to reduction of pulmonary hypertension or to blockade of direct effects of catecholamines on myocardial fibers or both.


The Canadian health care system: a model for American to emulate? The American health care system has the world's highest per capita costs and over 30 million citizens uninsured. The neighbouring Canadian system provides coverage for all basic medical and hospital services, at costs per capita that are about US$700 lower. Single-agency public funding allows tighter control of Canadian expenditures, and reduces administrative overheads. Hospitals are run as non-profit private corporations, funded primarily by a fixed annual allocation for operating costs. Most physicians are in private fee-for-service practice, but cannot charge more than the insured tariff negotiated between their provincial government and medical association. This approach, while attractive in its decentralization, tends to separate the funding and management of clinical services. Thus, hospital information systems lag a decade behind the USA, managed care initiatives are few, health maintenance organisations do not exist, and experimentation with alternative funding or delivery systems has been sporadic. Strengths of the system compared to the USA include: higher patient satisfaction, universal coverage, slightly better cost containment, higher hospital occupancy rates, and reduction in income-related rationing with more equitable distribution of services. Weaknesses in common with the United States are: cost escalation consistently outstripping the consumer price index with costs per capita second highest in the world, ever rising consumption of services per capita, inadequate manpower planning and physician maldistribution, poor regional co-ordination of services, inadequate quality assurance and provider frustration. Additional weaknesses include: an emerging funding crisis caused by the massive federal deficit, less innovation in management and delivery of care as compared to the USA, implicit rationing with long waiting lists for some services, and recurrent provider-government conflicts that have reduced goodwill among stakeholders. Thus, while the Canadian model has important advantages, it does not offer a panacea for American health care woes.


Screening for diabetic retinopathy: a relative cost-effectiveness analysis of alternative modalities and strategies. Diabetic retinopathy is the most common cause of blindness among adults of working age in the UK. If the disease is detected early effective treatment can be provided and this has resulted in calls for a systematic national screening programme. Using data on the screening of 3423 diabetics collected as part of an experimental programme in three UK centres, the relative cost-effectiveness of various screening options is assessed. The paper utilises direct evidence on a number of single modality screening options, including ophthalmoscopy undertaken by general practitioners or ophthalmic opticians, and non-mydriatic photography. With the objective of increasing the sensitivity of screening and using data collected in the study, options based on two further potential screening strategies are modelled and evaluated: combined screening using both ophthalmoscopy and non-mydriatic photography; and selective screening where high-risk diabetics are directly referred to an ophthalmologist and low-risk cases are either left unscreened or are screened by one of the single or combined modality screening options. Given the objective of early detection, effectiveness is assessed in terms of the sensitivity and specificity of the referral decisions of screening options. Both health service and private resource costs of the various screening options are estimated, the latter in terms of travel and the opportunity cost of time. Cost effectiveness is evaluated in terms of the expected cost per true positive case of diabetic retinopathy referred by the screening options. To narrow the choice between the options, those subject to three-way domination with respect to the three choice variables of sensitivity, specificity and expected cost per true positive are excluded. Amongst the remaining options, the choice is dependent on the trade-off between the higher specifics of unselective single modality screening options and the higher sensitivities and lower expected costs per true positive case detected of combined modality and selective screening options.


Cost savings in mass population screening for colorectal cancer resulting from the early detection and excision of adenomas. The widely-accepted hypothesis of a development sequence from colorectal adenoma to carcinoma is felt by clinicians to legitimate adenoma excision during routine colonoscopic investigation. Using published data on adenoma development, and adenoma prevalence data derived from the Nottingham colorectal cancer screening trial, the number of carcinomas prevented by early excision as a result of screening is predicted. The cost-effectiveness of early excision is then evaluated with reference to the expected treatment costs saved. These cost savings are found to represent a discount on the overall costs of mass population screening for colorectal cancer.


The morality of efficiency in health care--some uncomfortable implications. There are some general considerations which have implications for the delivery and finance of health care in all countries, not only Canada and the USA. Beginning with two propositions: that access to health care is a right of citizenship, which should not depend on individual income and wealth; and that the objective of health services is to maximise the impact on the nation's health of the resources available; the paper examines the ethical justification for pursuing efficiency in health care provision. The different meanings of efficiency are discussed in detail, and the use of quantitative indicators of health benefit, such as the QALY, placed in context. It is argued that the determination of health care resource allocations should take account of costs at both the macro planning level and the micro level of the individual doctor-patient relationship. Given the starting points the overall conclusion is that it is ethical to be efficient, since to be inefficient implies failure to achieve the ethical objective of maximising health benefits from available resources.


Discounting and health benefits.

Discounting and health benefits. This paper argues that non-monetary health benefits should not be discounted at the same rate as variables expressed in monetary terms. It argues instead that the appropriate discount rate should be at or close to zero. It explores the various influences of rising income, age and pure time preference on the relative value of current and future health states. It examines various arguments advanced to justify the current practice of discounting health benefits at the same rate as monetary costs. These include uncertainty and delay. The article concludes with an analysis of the likely impact of adopting a zero discount rate on the ranking of health interventions.


Discounting and health benefits: another perspective. This paper reviews the argument advanced by Parsonage and Neuburger that the non-monetary benefits of health programmes should be discounted at a lower rate than that used for financial flows. The conceptual issues raised in that paper are discussed and others, such as the tradability of non-monetary benefits and the link between individual and social discount rates, are introduced. The collection and assessment of more evidence is needed before Parsonage and Neuburger's proposition can be supported.


Non-culturable Aeromonas salmonicida in lake water. The survival of Aeromonas salmonicida subsp. salmonicida was investigated in lake water. During a 21-day study A. salmonicida became non-culturable in sterile lake water held at 10 degrees C. The incubation of replicate samples between 5 degrees C and 25 degrees C produced similar results. The recovery of colony-forming units of A. salmonicida from different lake water systems indicated that they survived longer in water that was naturally enriched (eutrophic) or enriched with tryptone soya broth. Flow cytometry, fluorescence light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) indicated that non-culturable cells were present. These cells could not be revived in dilutions of tryptone soya broth, whole dead fish or dissected fish tissue. Although viability could not be proven, it was shown that the morphological integrity found in viable cells was also maintained in non-culturable cells.


The effect of microcosm design on the survival of recombinant Pseudomonas putida in lake water. The survival of Pseudomonas putida marked with the xylE gene was monitored in lake-water microcosms. Various designs of microcosms were compared. These ranged from 250-ml conical flasks containing 100 ml surface lake water to 12-1 glass containers with lake water overlying sediment, continuous aeration and a supply of fresh surface lake water. The presence of a low flow-through rate was shown to have little effect on the survival of P. putida. An increase in the size of microcosm, presence of sediment and aeration had a significant effect on survival in lake water and increased the rate of decline of released cells. The implication of these results in predicting the survival of P. putida in lake water using microcosms is discussed.


Colonization of the digestive tract of germ-free mice by genetically engineered strains of Lactococcus lactis: study of recombinant DNA stability. The ability of genetically engineered Lactococcus lactis strains to become established in the digestive tract (DT) of germ-free mice was examined together with the stability of their genetic markers. Seven L. lactis strains were genetically modified by insertion of genetic markers on different replicons: chloramphenicol resistance gene cat was carried by self-transmissible plasmid pIL205, a derivative of plasmid pIP501; erythromycin resistance gene erm, originating from pAM beta 1, was inserted into non-transmissible plasmids pIL252 and pIL253 of low and high copy number respectively; erm gene from plasmid pMS1.5B was inserted into the chromosome. All strains carried a common wild-type plasmid pIL9 involved in lactose fermentation. It was observed that the DT of mice was rapidly and efficiently colonized with either the inoculated parental strain or with its derivatives or with both of them, but plasmid-free derivatives were always at dominant levels. Both plasmids pIL9 and pIL205 were lost, but the parental strains and the plasmid-lacking derivatives were at codominant levels, indicating that there is an equilibrium between plasmid loss and plasmid transfer in the DT. Strains that carried non-transmissible and low copy number plasmid pIL252 were rapidly eliminated from the DT, which in turn was colonized with the respective pIL252-less derivatives; this is probably due to the high segregational instability of pIL252.(ABSTRACT TRUNCATED AT 250 WORDS)


Isolation of astaxanthin over-producing mutants of Phaffia rhodozyma and their fermentation kinetics. Phaffia rodozyma CCRC-21346, CBS-6938, and CBS-5908 were treated with mutagenic agent NTG (N-methyl-N'-nitro-N-nitrosoguanidine) several times, then they were plated onto yeast-malt agar containing beta-ionone as a selective medium. Several isolates had increased astaxanthin content compared with the parental natural isolates. NCHU-FS301, one of the NTG treated strains, produced considerably more astaxanthin (1515.63 micrograms/g yeast) than the parent CBS-6938 (565.08 micrograms/g yeast). In studying the effects of carbon sources on the red pigment formation, it was found that glucose supported the highest total astaxanthin production (7809.3 micrograms/l). Yeast extract was the best nitrogen source in supporting the highest total astaxanthin formation (8637.5 micrograms/l). Beef extract, yeast extract, and potassium nitrate added in an equal ratio as a nitrogen source supported more pigment formation (8052.6 micrograms/l) than the rest of the mixture tested. Using kinetic parameters of specific growth rate (mu) and specific astaxanthin productivity (qp) to judge the association between growth behavior and product formation, the NCHU-FS301 showed more positive growth-associated fermentation type than the parent strain. These astaxanthin-overproducing mutants could be useful for the aquacultural industry in providing a natural source of astaxanthin.


Effect of moisture content on the microbial activity in JP-5 fuel oil. The water solubility of JP-5 fuel oil was found to be proportional to the reciprocal of absolute temperature from 0 degree C to 60 degrees C. Water in the fuel oil would become condensed once the temperature was shifted from a high temperature to low temperature. During the storage, condensed water was precipitated in the bottom of the tank. Both the static and the dynamic dehumidification processes with molecular sieve could reduce the moisture content of fuel oil to less than 5 ppm. However, pre-dried fuel oil had a mildly hydroscopic phenomenon at relatively high humidity condition. The spores of contaminated microbes could survive in the fuel oil with 5 to 80 ppm of moisture content. High moisture content of fuel oil was not favorable to the spore survival. Penicillium sp. could survive in the fuel oil longer than Cladosporium resinae.


Correlation of tumor necrosis factor alpha levels with disease activity of rheumatoid arthritis. In this study, positive correlations were found between tumor necrosis factor alpha (TNF alpha) levels in the sera of rheumatoid arthritis (RA) patients and the duration of morning stiffness, joint tenderness count, the Ritchie articular index and erythrocyte sedimentation rate. Further, higher mean grades of disease activity of RA were accompanied by correspondingly higher levels of serum and synovial TNF alpha. In the longitudinal study, when the disease activity of RA decreased after treatment, serum TNF alpha levels also decreased. This suggests that the levels of serum and synovial TNF alpha correlate positively with RA disease activity. The levels of TNF alpha in synovial fluid were significantly higher in RA and acute gouty arthritis (GA) patients than in osteoarthritis (OA) patients. This suggests that joint inflammation in inflammatory arthritis related to local production of TNF alpha in the joint cavities. Serum TNF alpha levels in RA patients were significantly higher than those in the OA patients; no statistical difference was found between acute GA and OA patients.


Evaluation of anti-HTLV-1 in physical-checkup individuals and blood donors in Taiwan. The gelatin particle agglutination (GPA) test specific for antibody detection of human T-cell lymphotropic virus type 1 (HTLV-1) was used to screen 500 blood donors and 5000 physical-checkup individuals at Veterans General Hospital-Taipei. The positive rate of physical-checkup individuals and the blood donors was 0.18% (9/5000) and 0% (0/5000) respectively. Among the 9 GPA positive specimens, eight were confirmed to be positive by western blot analysis and a prevalence rate of 0.16% (8/5000). Seven of the nine GPA positive samples were also positive by indirect fluorescent antibody test and two of them had indeterminate results. Since GPA is less expensive, relatively simple and convenient, we recommend that GPA could be used as screening test for HTLV-1 infection of blood donors, followed by western blot method as a confirmatory test in blood bank.


[Evaluation of biological toxicity of livestock fecal deordoring microbial agents]. Eight commercial livestock microbial agents were selected for the evaluation of biological toxicity. The general physical, chemical and microbial analyses of these microbial agents were performed and the acute toxicity of mice, aquatic living organisms and the germinated inhibition of vegetable seeds by these microbial agents were also estimated for the evaluation of biological toxicity. The results have shown that all livestock microbial agents have no pathogenic Salmonella and Staphylococcus aureus been detected. However, the acute biological toxicity test of water extractant of these microbial agents indicated that microbial agent A, B, C, D and M appeared various toxicity to the mice. The dosage of LC50 (48 hr) to the mice was 3 mg/ml for microbial agent A (150 mg/kg mouse, high toxicity), 90 mg/ml for microbial agent B (4.5 g/kg mouse, low-medium toxicity), 92 mg/ml for microbial agent C (4.6 g/kg mouse, low-medium toxicity), 118 mg/ml for microbial agent D (5.9 g/kg mouse, low toxicity) and nontoxicity for microbial agent M, respectively. In addition, the dosage of LC50 to aquatic organisms Daphina similis and Lemna minor, was < 0.01 mg/ml and 0.13 mg/ml for microbial agent A, 1.73 mg/ml and 10 mg/ml for microbial B, 3 mg/ml and > 20 mg/ml for microbial C, 0.56 mg/ml and 14 mg/ml for microbial agent D, respectively. All microbial agents also show various inhibitory effects to the seed germination and root extension of vegetable crops. The study has demonstrated that the biological toxicity of livestock microbial agents was following such order: microbial A > B, C > D > M. The animal has shown more sensitive than plant to the microbial agents.


Cryptococcus albidus isolated from pigeon excreta in Taiwan. Cryptococcus albidus (Saito) Skinner, found frequently in the air of El-Minia, Egypt; in New Zealand and Budapest, and isolated occasionally from clinical specimens, has now been recognized as the cause of human diseases. Since isolation of this fungus from pigeon droppings has not previously been reported, this first paper reports results of its drug-susceptibility testing to gentamicin and 5-fluorocytosine, as well as its fungal cell antigenic cross reactivity with C. neoformans as detected by a Latex-Crypto-Antigen Detection System (LCAS).


[Long term and low dose treatment with methotrexate in corticosteroid-dependent asthma--two-year clinical experience]. Ten patients with corticosteroid-dependent asthma were treated with long-term and low dose methotrexate (MTX) for its corticosteroid-sparing effect. The average age was 51.2 years (ranged 24 to 67). Three were women. Despite the use of maximal doses of bronchodilators, their daily prednisolone dosages were always more than 10 mg during an average period of 2.75 years (ranged 1 to 6 years). Following the use of oral MTX, 15mg weekly from more than 6 months (averaged 11.8 months; ranged 6 to 15 months), the average daily requirement of prednisolone decreased from 14.5 to 6.5 mg (p < 0.01). Among them, four did not need steroid and the other six had a marked subjective improvement in breathing, cough and nocturnal dyspnea. However, three of them could not have steroid dose reduced. As for adverse reactions to MTX in ten patients, two patients had nausea and vomiting, two had skin eruption, three had somnolence, and one had elevated sGOT (78 U/L). The adverse effects were all transient. Neither oral ulcer, nor leukopenia was found among them. This study suggests long-term low dose oral MTX may have a steroid-sparing effect in steroid-dependent asthmatic patients. Their adverse effects were mild and transient.