Polymerase chain reaction for detection of Mycobacterium tuberculosis. Rapid diagnosis of tuberculosis is essential, and therefore we use a polymerase chain reaction. In this report, we describe two cases of tuberculous lymphadenitis in childhood. Although histopathological findings were not specific for tuberculosis in both cases, distinct positive bands were amplified. For DNA diagnosis of tuberculosis, a lysis method of extracting chromosomal DNA from lipid-rich cell walls of mycobacteria is of critical importance. We made use of a simple lysozyme-proteinase K treatment for biopsied tissues. Although this extraction procedure was less efficient than those reported previously, it was considered sufficient for detecting mycobacterial DNA with the use of a highly sensitive polymerase chain reaction. We conclude that DNA amplification in combination with lysozyme lysis can be used routinely in clinical laboratories as a rapid and sensitive test for the diagnosis of tuberculosis.


Evaluation of captopril versus reserpine and frusemide in treating hypertensive children with acute post-streptococcal glomerulonephritis. We have evaluated the effect of oral captopril versus a combination of oral reserpine and frusemide in the treatment of 20 children with post-streptococcal glomerulonephritis (APGN) with hypertension. Captopril produced a significantly greater reduction in systolic and diastolic blood pressure in both the standing and supine positions than reserpine + frusemide at 0.75, 1, 1.25, 8, 24, 48 and 72 h after initiating treatment. Neither postural hypotension nor reflex tachycardia accompanied the therapeutic effect of captopril. Blood urea, serum creatinine and creatinine clearance did not change significantly after therapy in either study group. Three days after initiating treatment, the 24 h urinary catecholamine output increased significantly in children who received captopril but did not change in children treated with frusemide and reserpine.


High density lipoprotein cholesterol and total cholesterol in children with asthma and allergic rhinitis. Serum high density lipoprotein cholesterol and total cholesterol were studied in 15 children with allergic rhinitis, 45 asymptomatic asthmatic children and 16 children with acute asthmatic attacks. The latter were also studied in their asymptomatic phase. Two control groups of children with minor anatomical defects and those with acute pneumonia with respiratory distress were also studied. The serum concentrations of high density lipoprotein cholesterol were significantly higher (p less than 0.001) and those of total cholesterol lower (p less than 0.001) in children with respiratory allergy when compared to controls. The concentration of high density lipoprotein cholesterol increased and that of total cholesterol decreased during acute asthma.


A follow-up study of the diet of Finnish diabetic adolescents. Changes with age and time in energy-adjusted food consumption and nutrient intake of 74 diabetic subjects initially aged 12-17 years were studied. Food consumption was measured by the 48-h recall method. During the three-year follow-up (from 1985 to 1988), the proportion of carbohydrate of total energy intake decreased from 49% to 47%, that of fat increased from 33% to 36% and that of protein decreased slightly. The densities of fibre and several vitamins decreased in the diet of the diabetic adolescents. These unfavourable changes in the diet of diabetic adolescents took place with increasing age and duration of diabetes, while virtually no changes with time were detected.


Collagen type II in Langer-Saldino achondrogenesis: absence of major abnormalities in a less severe case. Collagen extracted either from cartilage or synthesized in vitro was analyzed to identify possible molecular defects in the cartilaginous matrix of a male fetus suffering from a mild form of type II achondrogenesis (Langer-Saldino). The tissue architecture of the patient's cartilage was markedly altered and showed numerous fibrous vascular canals which were focally stained by antibodies against collagens I and III. Collagen II was present, although heterogenously distributed throughout the cartilaginous matrix. Upon electrophoretic separation, however, the patient's femoral head cartilage showed the presence of collagens II, IX and XI only, which was similar to an age-matched control. The hydroxyproline/hydroxylysine ratio of collagen II of the patient was not significantly different from that of the control. Likewise, the compositions of collagens synthesized by cultured chondrocytes as well as fibroblasts were similar in the patient and the control. The results provide strong evidence that, in the present mild case of Langer-Saldino achondrogenesis, collagen II is expressed and regularly hydroxylated at its lysyl residues. This may indicate that cartilage components other than collagen II may be responsible for the altered tissue organization observed. Along with previous observations, our data suggest that the degree of biochemical matrix alterations may be related to the severity of the clinical phenotype.


Oral zinc as initial therapy in Wilson's disease: two years of continuous treatment in a 10-year-old child. Two years of continuous therapy promoted a significant overall amelioration in a 10-year-old boy affected by an hepatic form of Wilson's disease in which zinc sulphate was the sole therapy. In particular, liver function returned to normal and hepatic histology also improved. The parameters characterizing copper metabolism were kept under good control, and a decrease in copper concentration was found in both erythrocytes and liver. The copper balance study performed during the 25th month of treatment showed that oral zinc was still efficiently inhibiting the intestinal absorption of copper. No side effects have been reported so far.


Rett syndrome: the late infantile regression period--a retrospective analysis of 91 cases. The late infantile regression period of Rett syndrome was analysed in a consecutively collected series of females, born 1945-87 (median 17 1/2 years), fulfilling the diagnostic criteria for classic Rett syndrome. For general information, data from 91 girls and women were used (group A), while more detailed analysis was based on the youngest 20, all born during the last decade (group B). Median ages at onset of developmental stagnation (stage I) were 11 (5-24) and 10 (5-18) months for groups A and B, respectively. Loss of acquired skills (stage II) began at 19 (12-36) and 17 (13-25) months, and lasted for 19 (2-53) and 13 (2-32) months. Onset of the period of loss was distinctly marked in 43%, dramatic in 16% and insidious in 41%. Gross motor delay and aberrant behaviour raised the first suspicions of disease, while a more specific pattern comprising a triad of manifestations, i.e. contact/communication, hand use/skill, babble/words, characterized the loss of acquired skills. Although development invariability came to a definite break at a crucial stage of maturation, an "awakening" (stage III) and return of interest to act and interact astonishingly followed the period of withdrawal. Apraxia was found in all, but some use of the hands could be enticed in engaging and non-demanding situations. The observed preference for use of the left hand (9/11), compared to the right (2/11), suggested a reflection of an underlying dysfunction/disease process, also displayed as spike activity in left central leads in EEGs from 13/20 girls.


Influence of surgical treatment of scoliosis in children with spina bifida on ambulation and motoric skills. The influence on motoric skills of surgical treatment of scoliosis in 14 children with myelomeningocele was studied. Fifty percent of the children had increased flexion contractures of the hips at follow-up, and all but one patient had impaired motor ability. There was no influence on activities of daily life, but 57% of children had lost some of their ambulation capacity. Postoperative physiotherapy is highly advisable. Intensive attempts to treat these children conservatively to prevent scoliosis progression is suggested.


Congenital dislocation of the hip: a prospective study comparing ultrasound and clinical examination. Screening for congenital dislocation of the hip by standard clinical methods and ultrasound was compared prospectively in 1503 newborns (1291 girls and 212 boys). A total of 82.8% of the hips (78.8% of the infants) had well developed acetabulae, 14.5% (17.2% of the infants) were shallow (immature) and 2.7% (4.1% of the infants) were dysplastic. Within 1-3 months 96.7% of the infants with clinically stable, but immature hips normalized, while 3.3% did not improve or worsened, and therapy was therefore initiated. Indications for treatment included dysplastic and/or clinically unstable hips, and a higher number of newborns were treated during the study period than in 1982-85 (31.2 vs 19.7 per 1000, p = 0.0002). Thirty-seven percent of the patients had both clinical and ultrasound findings compatible with congenital dislocation of the hip, while the decision to treat was based on clinical findings alone in 25.0% and on ultrasound findings in 38.0%. Infants born during the study period of nine months had a low incidence of late congenital dislocation of the hip compared with our earlier reported results from 1982-85 (0.9 vs 3.5 per 1000, p = 0.012). The study demonstrated major discrepancies between clinical and ultrasound evaluation of hips in the newborn, and the role of ultrasound in the screening for congenital dislocation of the hip requires further evaluation.


Transverse myelitis following mumps in children. Although acute transverse myelitis is a rare complication of mumps, it is relatively well documented. We describe a child who developed mumps associated acute transverse myelitis and who subsequently recovered completely. To our knowledge, only 13 cases have been reported in children. This case is compared with 13 previously reported patients.


Precision of gestational age assessment in the neonate. The precision of the Ballard scale for assessing gestational age (GA) was evaluated in a consecutive sample of 227 preterm and/or low-birth-weight neonates. Each newborn was rated independently by two neonatologists and the difference in GA estimation between them was computed. The estimated precision was not high, the 95% tolerance interval estimate being as large as 7.4 weeks. The precision of the neurologic and physical parts of the scale was poorer than that of the complete scale (95% of differences less than 10.5 and 9.2 weeks respectively), and more influenced by the type of delivery. These findings are not unexpected from statistical theory, and cast doubts on the use of only the physical part of the Ballard scale in assessing GA, since greater accuracy could be accompanied by reduced precision.


Exposure assessment for electric and magnetic fields. Exposure assessment for extremely low frequency (ELF) electric and magnetic fields (EMF) is discussed. It is suggested that such assessments can be designed by attempting to mimic the receptor's experience of interest. Present efforts are, however, hampered by the lack of clearly defined human health effects, or even important interaction mechanisms, which might be used to define the appropriate exposure measure, the experience of interest. The state of the art of ELF EMF exposure assessment is described, including the use of surrogates, models, and available instrumentation. Data indicate that magnetic field levels in the home are of the order of 0.1 microT and that nonoccupational total exposures may best be predicted by levels in the bedroom. Occupational exposures may be orders of magnitude higher where high currents are present.


[Problems in clinical research].

[Problems in clinical research]. In recent years, the requirements of society towards the quality of research have increased. Especially in the perspective of European integration, ethical and conceptual problems have to be revisited. Clinical research is alimented by technical development and the results of fundamental and pharmacological research. The obligation of converting the most recent knowledge into conventional treatment is the driving force of development. Clinical research is fraught with multiple risks, mainly the risks incurred by patients enrolled in clinical studies, and the risks of drawing erroneous conclusions from badly designed studies. This is the reason why the organization of clinical studies must satisfy the highest quality standards. Prospective randomized double-blind studies are doubtless the best design although they usually involve the difficult problem of patient information. As concerns publications, the picture may be distorted. Thus all the studies should be published exhaustively. Clinical research is thus a great challenge from both the intellectual and the ethical points of view.


[Lymphoscintigraphy with (123)I-marked epidermal growth factor in cervix cancer]. Several malignant neoplasms express high levels of epidermal growth factor receptors. The aim of this study was to assess whether 123I-labeled epidermal growth factor can concentrate in lymph node metastases of squamous cell carcinomas of the cervix. 14 patients with advanced cervical cancer were selected because of their high probability of lymph node metastases. Planar scintigrams were recorded from the lower and upper abdomen following subcutaneous injection of 123I-labeled epidermal growth factor into the web space of each foot. Scintigraphic images were interpreted without knowledge of computerized tomography scan (n = 13) and ultrasound (n = 9) results from the pelvic lymph nodes. In 2 patients, histological verification was performed by diagnostic biopsy of pelvic lymph nodes. Nodal involvement was confirmed by computerized tomography for 4 of the 11 positive scans and by ultrasound for 2. In 11 out of 14 patients an increased uptake of 123I-labeled epidermal growth factor could also be seen in the primary tumour. Our findings suggest that targeting of cervical cancer lymph node metastases can be achieved by in vivo binding of 123I-labeled epidermal growth factor with receptors on tumour cell surfaces.


Validation of a terrestrial food chain model. An increasingly important topic in risk assessment is the estimation of human exposure to environmental pollutants through pathways other than inhalation. The Environmental Protection Agency (EPA) has recently developed a computerized methodology (EPA, 1990) to estimate indirect exposure to toxic pollutants from Municipal Waste Combuster emissions. This methodology estimates health risks from exposure to toxic pollutants from the terrestrial food chain (TFC), soil ingestion, drinking water ingestion, fish ingestion, and dermal absorption via soil and water. Of these, one of the most difficult to estimate is exposure through the food chain. This paper estimates the accuracy of the EPA methodology for estimating food chain contamination. To our knowledge, no data exist on measured concentrations of pollutants in food grown around Municipal Waste Incinerators, and few field-scale studies have been performed on the uptake of pollutants in the food chain. Therefore, to evaluate the EPA methodology, we compare actual measurements of background contaminant levels in food with estimates made using EPA's computerized methodology. Background levels of contaminants in air, water, and soil were used as input to the EPA food chain model to predict background levels of contaminants in food. These predicted values were then compared with the measured background contaminant levels. Comparisons were performed for dioxin, pentachlorophenol, polychlorinated biphenyls, benzene, benzo(a)pyrene, mercury, and lead.


Regional differences in dietary intake of environmental pollutants in Japan. In order to study regional differences in the dietary intake of environmental pollutants in Japan, we measured the daily intake ingested via food (the main route for pollutant intake) and analyzed the data by multivariate analysis. Daily mean samples were collected by the duplicate method from 25 women per area in 5 cities in both northern and southern Japan in July, 1987. Cadmium, lead, benzo(a)pyrene, hexachlorobenzene, DDT, chlordane isomers, and the total amount of the isomers were determined for typical pollutants. The data were statistically analyzed by multivariate analysis. Regional differences in the daily intake of environmental pollutants were studied by discriminant analysis. The eigenvalues of the functions 1 and 2, which accounted for 84.8% of variance, were 0.647 and 0.221, respectively. The function 1 was related to regional differences in the intake of environmental pollutants in the northern and southern regions of Japan. The statistical distance calculated from the intake of environmental pollutants correlated well with the geographical distance. The coefficients of the canonical discriminant function 1 were large in the order of lead (Pb) and the total amount of chlordane isomers (T-CLD). The regional difference was assumed to be due to the intake of Pb in the north and the intake of T-CLD in the south. Other pollutants, however, did not contribute to the observed regional difference. The study shows that performing multivariate analysis by computer provides a useful means for processing a large amount of data and can provide important information for data analysis in environmental health.


The measurement of salivary cadmium by electrothermal atomic absorption spectrophotometry and its use as a biological indicator of occupational exposure. We have examined the potential use of salivary cadmium measurements for the biological monitoring of occupational cadmium exposure, paying particular attention to the contamination risks associated with such measurements. We have developed a method for the direct determination of cadmium in saliva by graphite furnace atomic absorption spectrophotometry, which minimizes the risk of contamination during sample preparation and analysis. The limit of detection is 0.6 nmoll-1, which is sufficiently sensitive to discriminate between unexposed and occupationally exposed individuals. The method has been employed to measure cadmium levels in saliva samples collected by two different methods from an unexposed population, a group of ex-workers previously exposed to cadmium (Group 1), and two groups of currently exposed workers (Groups 2 and 3). Salivary cadmium levels were significantly raised in both of the groups of currently exposed individuals (group 2 median (Md) = 17 nmoll-1, group 3 Md = 70 nmoll-1, p greater than 0.0001), and in past workers with previous long-term exposure (Group 1 Md = 2.5 nmoll-1, p greater than 0.001) when compared with an unexposed population. The results suggest that the measurement of salivary cadmium may reflect recent exposure to the metal. However, considerable care must be taken in collecting samples because a risk of contamination during sampling is apparent with procedures commonly used for saliva collection, and for this reason the applicability of such measurements for biological monitoring is limited.


[Early diastolic incisures in uteroplacental blood vessels. Experiences in patients with uneventful pregnancy and labor]. The frequency of diastolic notching in uteroplacental arteries was examined in a collective of 510 patients with uneventful pregnancies and deliveries. Notching appeared to be more frequent in the uterine than arcuate artery. It was seen to increase as pregnancy progressed and was markedly higher in the third compared with the second trimester. Uteroplacental notching occurred more frequently unilaterally and mainly affected contralateral sites of the placenta. Bilateral and retroplacental notching was the exception in normal pregnancies.


[Cytologic cancer prevention with smears of the uterine cervix and endocervical canal in pregnancy]. This study was performed to evaluate the importance of obtaining prepartum Papanicolaou smears routinely. 1,417 patients receiving pregnancy care had normal prenatal Pap smears. 88 (5.85%) had abnormal prepartum Pap smears. Thirteen (14.8%) of the abnormal smears showed squamous dysplasia, 2 (2.3%) showed an adenocarcinoma or squamous invasive carcinoma. The results of this study support the practice of performing Pap smears during prenatal care.


Wind velocity effects on sampling rate of NO2 badge. The effects of wind velocity on a sampling rate of a nitrogen dioxide (NO2) diffusive badge were experimentally determined using a turntable. The use of a turntable permits the collection of the large amounts of data that are needed for statistically reliable results at several wind velocities in one experiment. The regression model for the wind effect determined by these experiments was closely correlated with data previously gathered from experiments using wind tunnels. Experiments at two different relative humidities, 35% and 60%, were performed and analyzed by a simple least square regression model. A multi-regression model containing two independent variables, wind velocity and relative humidity, also was developed. The multi-regression model was useful at relative humidity between 20% and 60% and wind velocity between 0 and 7 meter per second (m/sec).


Management of sudden bereavement in the accident and emergency department. To assess facilities available for the suddenly bereaved in accident and emergency departments and variations in care of bereaved relatives. Postal questionnaire survey. England and Wales. All 98 accident and emergency departments treating over 50,000 patients a year, 78 of which replied. Number of departments with specific facilities, staff training, and procedures for dealing with bereavement. 60 hospitals had a specific room for bereaved relatives; the remainder used multipurpose rooms. In 49 hospitals relatives were taken to the room by a nurse with sole responsibility for caring for them. In 40 hospitals the nurse stayed with the relatives and 66 updated relatives on the patient's condition. Facilities for viewing the body privately were poor, and relatives often had to ask to be left alone. 25 departments gave no written information on bereavement and only four routinely followed up relatives. Further training was requested by staff in 44 departments. Although facilities could be improved, immediate care of relatives is good. Care over subsequent weeks and preparation for this period is invariably inadequate.


Audit of a tuberculosis contact tracing clinic. To determine the efficiency of tuberculosis contact tracing in South Glamorgan 1987-9. Review of records of contact tracing clinic and of data from the Mycobacterium Reference Unit. The clinic's practice was compared with 1983 British Thoracic Society's recommendations. Health authority tuberculosis control programme. Proportion of contacts screened, follow up attendance rates, number of secondary cases detected, and quality of record keeping. 101 index patients and 611 contacts were identified. 596 (97.5%) contacts were screened, of whom 139 should not have been. Of 356 contacts requiring a Heaf test, 237 were tested, seven refused the test, and 112 had chest radiography without a Heaf test. 95 contacts were unnecessarily tested. 87 contacts had chest radiography unnecessarily and seven should have had radiography but did not. 34 contacts were given follow up appointments inappropriately and seven were overlooked for follow up. Tuberculosis was diagnosed in five asymptomatic contacts, all at initial screening and all close contacts of index patients with pulmonary disease. Inadequacy of data, non-adherence to contact tracing guidelines, and failure to define the term highly infectious index case resulted in many contacts being unnecessarily screened or followed up. The efficiency of tracing contacts would be improved by specifying smear results and ethnic origin of the index case on the notification form, clearly classifying contacts as close or causal, and clearly defining the term highly infectious.


Must diabetes be a fatal disease in Africa? Study of costs of treatment. To estimate the costs of diagnosis and treatment of diabetes in Tanzania. Costs estimated from the reported and recorded experience of patients with newly presenting diabetes in 1989-90 and of diabetic patients first seen in 1981-2. Muhimbili Medical Centre, Dar es Salaam. 464 patients (315 men and 149 women). 262 patients diagnosed during 1 September 1989-31 August 1990 (group 1) and 202 during 1 June 1981-31 August 1982 (group 2). The average annual direct cost of diabetes care in 1989-90 was $287 for a patient requiring insulin and $103 for a patient not requiring insulin. Purchase of insulin accounted for 68.2% ($156) of the average annual outpatient costs for patients requiring insulin. For patients not requiring insulin the cost of oral hypoglycaemic drugs and treatment of chronic complications and infections accounted for 42.5% ($29.3) and 48.8% ($33.7) of costs respectively. Cost of outpatient care of diabetic patients for the whole of Tanzania was estimated at $2.7m, *75,128 (32.2%) of which was for insulin. Doctors' and nurses' costs accounted for 0.2% of total costs of outpatient care. The annual direct inpatient care costs were estimated at $1.25m. Around 0.2% of the Tanzanian population aged 15 years and over used the equivalent of 8% of the total government health expenditure, which was $47,4088,382. Diabetes places a severe strain on the limited resources of developing countries. If African patients with diabetes have to pay for their treatment most will be unable to do so and will die.


Hyponatraemia and death or permanent brain damage in healthy children. To determine if hyponatraemia causes permanent brain damage in healthy children and, if so, if the disorder is primarily limited to females, as occurs in adults. Prospective clinical case study of 16 affected children and a review of 24,412 consecutive surgical admissions at one medical centre. 16 children (nine male, seven female; age 7 (SD 5) years) with generally minor illness were electively hospitalised for primary care. Consultation was obtained for the combination of respiratory arrest with symptomatic hyponatraemia (serum sodium concentration less than or equal to 128 mmol/l). Presence, gender distribution, and classification of permanent brain damage in children with symptomatic hyponatraemia in both prospective and retrospective studies. By retrospective evaluation the incidence of postoperative hyponatraemia among 24,412 patients was 0.34% (83 cases) and mortality of those afflicted was 8.4% (seven deaths). In the prospective population the serum sodium concentration on admission was 138 (SD 2) mmol/l. From three to 120 inpatient hours after hypotonic fluid administration patients developed progressive lethargy, headache, nausea, and emesis with an explosive onset of respiratory arrest. At the time serum sodium concentration was 115 (7) mmol/l and arterial oxygen tension 6 (1.5) kPa. The hyponatraemia was primarily caused by extrarenal loss of electrolytes with replacement by hypotonic fluids. All 16 patients had cerebral oedema detected at either radiological or postmortem examination. All 15 patients not treated for their hyponatraemia in a timely manner either died or were permanently incapacitated by brain damage. The only patient treated in a timely manner was alive but mentally retarded. Symptomatic hyponatraemia can result in high morbidity in children of both genders, which is due in large part to inadequate brain adaptation and lack of timely treatment.


Visual problems in the elderly population and implications for services. To determine the prevalence of visual disability and common eye disease among elderly people in inner London. Cross sectional random sample survey. Inner London health centre. Random sample of people aged 65 and over taken from practice's computerised age-sex register. Presenting binocular Snellen 6 m distance acuity and best monocular 3 m Sonksen-Silver acuity to classify prevalence of blindness by World Health Organisation criteria (less than 3/60 in better eye) and American criteria for legal blindness (better eye equal to 6/60 or less) and of low vision by WHO criteria (best acuity 6/18) and visual impairment by American criteria (less than 6/12 or 20/40 but greater than 6/60 or 20/200 in better eye). Principal cause of visual loss by diagnosis, referral indication by cause to hospital eye service, and proportion of cases known to primary care. 207 of 288 (72%) eligible people were examined. 17 (8%) housebound subjects were examined at home. The prevalence of blindness was 1% by WHO criteria and 3.9% by American criteria. The prevalence of low vision (WHO criteria) was 7.7%. The prevalence of visual impairment (American criteria) was 10.6%. Cataract accounted for 75% of cases of low vision. Only eight out of 16 patients with low vision were known by their general practitioner to have an eye problem. 56 subjects (27%) would probably have benefited from refraction. Comparisons with studies in the United States and Finland suggested higher rates in this sample, mainly due to the prevalence of disabling cataract. There seems to be a considerable amount of undetected ocular disease in elderly people in the community.


Effect of unexpected demolition announcement on health of residents. To examine the impact of an unexpected announcement of the demolition of a housing estate on the health of the area's residents. Study of general practitioner consultation rates of the estate's residents after the announcement compared with those of other areas and with those of the previous year. General practices in Runcorn, Cheshire. 17,000 patients on lists of the two group practices serving the estate and surrounding area. Relative weekly consultation rates with general practitioners. The mean adjusted odds ratio for consultation was 1.12 (SD 0.12) when demolition was expected and 0.877 (0.05) when it was not (t = 5.94, p less than 0.001). The difference remained after the adjustment for the fall in the estate's population was removed (t = 3.7, p less than 0.01). Announcement of the estate's demolition adversely affected residents' health.


Meningococcal infections: reducing the case fatality rate by giving penicillin before admission to hospital. To determine whether parenteral penicillin given before admission to hospital reduces the case fatality rate in patients with meningococcal disease. Retrospective analysis of 46 consecutive patients admitted to hospital with meningococcal disease from January 1986 to March 1991. District general hospital. Hospital case fatality rate. None of the 13 patients given parenteral penicillin by the referring doctor before admission died, compared with eight deaths (24%) in 33 patients admitted without such treatment. Parenteral penicillin given before admission probably contributed to a reduction in the case fatality rate from meningococcal disease, and primary care physicians should be encouraged to give such treatment immediately on suspicion of the diagnosis before transferring the patient to hospital. Public health physicians are well placed to inform and alert general practitioners of the potential benefit of this action.


Early treatment with parenteral penicillin in meningococcal disease. To measure the effect of parenteral antibiotics given before admission to hospital on mortality and on bacteriological investigations in meningococcal disease. Retrospective review of hospital notes and laboratory and public health medicine department records. Three health districts in south west England. Patients with meningococcal disease in Gloucester district presenting between 1 January 1982 and 31 December 1991 (n = 190); patients with meningococcal disease in Plymouth (n = 118) and Bath (n = 73) districts presenting between 1 January 1988 and 31 December 1991 (total = 381). Number of deaths from meningococcal disease. Parenteral antibiotic given by general practitioners was associated with a substantial reduction in mortality (from 9% to 5%; relative risk 0.6, 95% confidence interval 0.2 to 1.5); patients with a rash were more likely to be given parenteral antibiotics, and mortality was further reduced (from 12% to 5%; 0.5, 0.2 to 1.4). In a district where such treatment was regularly encouraged its use increased from 5% to 40% of cases over 10 years (p = 0.00001). Treatment with parenteral antibiotics before admission made isolation of meningococci from blood and cerebrospinal fluid less likely but did not affect nasopharyngeal cultures. General practitioners should carry benzylpenicillin in their emergency bags at all times and should administer it promptly, preferably intravenously, whenever meningococcal disease is suspected, unless the patient has had an anaphylactic reaction to penicillin. Specimens for culture should include a nasopharyngeal swab.


Follow up of patients presenting with fatigue to an infectious diseases clinic. To determine the symptomatic and functional status during follow up of patients referred to hospital with unexplained fatigue and to identify patient variables associated with persistent functional impairment. Follow up by postal questionnaire six weeks to four years (median 1 year) after initial clinical assessment of patients referred to hospital during 1984-8. Infectious diseases outpatient clinic in a teaching hospital. 200 consecutive patients with fatigue of uncertain cause for at least six weeks; 177 fulfilled the inclusion criteria. Findings at initial assessment; current symptoms, beliefs about the cause of illness, coping behaviours emotional disorder, social variables including membership of self help organizations, and degrees of recovery and functional impairment from questionnaire responses. 144 (81%) patients returned completed questionnaires. Initial assessment did not indicate the cause of fatigue, other than preceding infection. The proportion of patients with functional impairment was significantly smaller with longer follow up (33% (11/33) at two to four years, 73% (29/40) at six weeks to six months; chi 2 for trend = 12.5, df = 1; p less than 0.05). Functional impairment was significantly associated with belief in a viral cause of the illness (odds ratio = 3.9; 95% confidence interval 1.5 to 9.9), limiting exercise (3.2; 1.5 to 6.6), avoiding alcohol (4.5; 1.8 to 11.3), changing or leaving employment (3.1; 1.4 to 6.9), belonging to a self help organization (7.8; 2.5 to 23.9), and current emotional disorder (4.4; 2.0 to 9.3). Short term prognosis for recovery of function was poor but improved with time. Most patients had made a functional recovery by two years after initial clinic attendance. Impaired functioning was more likely with certain patient characteristics. Prospective studies are required to clarify whether these associations are the consequences of a more disabling illness or indicate factors contributing to impaired function.


Association between postpartum thyroid dysfunction and thyroid antibodies and depression. To define the relation between mood and autoimmune thyroid dysfunction during the eight months after delivery. Double blind comparison of the psychiatric status of women positive and negative for thyroid antibodies. Clinical examination and blood sampling for free triiodothyronine and thyroxine, thyroid stimulating hormone, and thyroid antibody concentrations at four weekly intervals. Psychiatric assessment at six, eight, 12, 20, and 28 weeks post partum. Outpatient department of district hospital. 145 antibody positive women and 229 antibody negative women delivering between August 1987 and December 1989. Thyroid status. Number of cases of mental ill health by the general health questionnaire, research diagnostic criteria, Hamilton 17 item depression scale, hospital anxiety and depression scale, and Edinburgh postnatal depression scale. Six weeks after delivery the general health questionnaire showed 62 (43%) antibody positive women and 65 (28%) antibody negative women had mental ill health (chi 2 = 8.18, p less than 0.005). Follow up of 110 antibody positive and 132 antibody negative women showed significantly greater depression by research diagnostic criteria in antibody positive women (47%) than antibody negative women (32%) regardless of thyroid dysfunction. Antibody positive women showed higher mean scores for depression on the Hamilton (6.01 v 3.89, p = 0.0002), Edinburgh (7.45 v 5.92, p = 0.031), and hospital depression scales (4.95 v 3.79, p = 0.003). Depressive symptoms are associated with positive thyroid antibody status in the postpartum period.


Effect of reduced temperatures and brefeldin A on prolactin secretion and on subcellular distribution of the secretory product and membrane antigens in GH3 pituitary cells. The effects of reduced temperatures (20, 15 or 10 degrees C) and brefeldin A (BFA) on prolactin (PRL) secretion in the GH3 rat pituitary cell line have been compared. Both treatments inhibit PRL release to different extents. Ultrastructural immunocytochemistry reveals that, depending on the treatment, PRL is blocked at different steps during its intracellular transit. The temperatures of 20 and 15 degrees C block the PRL transport at one face of the Golgi stacks whereas both the temperature of 10 degrees C and BFA treatment induce an arrest of PRL at the level of the rough endoplasmic reticulum (RER) cisternae. Moreover, exposure to 10 degrees C or BFA induces an accumulation of a specific Golgi membrane antigen in the dilated RER structures. However, although disorganized and no longer definable under BFA treatment, the Golgi apparatus remains visible at 10 degrees C. These two last treatments cause also an increase in the number of partly rough, partly smooth tubular structures tentatively called 'paired cisternae'.


Distribution of intramembranous particles and filipin-sterol complexes in the spermatid and spermatozoon of Culex quinquefasciatus (Culicidae). A freeze-fracture study was carried out on spermatid and spermatozoon of the mosquito Culex quinquefasciatus. In the spermatid plasma membrane few and randomly distributed intramembranous particles were observed. In the spermatozoon the density of intramembranous particles was higher on the P- than on the E-fracture face of the plasma membrane. Two populations of particles were observed. Large particles (about 15 nm in diameter) are regularly arranged in double rows as a zipper-line, longitudinally oriented in relation to the main cell axis. These strands of particles were observed in the posterior head region, mainly associated with the E-fracture face. Filipin was used to analyse the presence and distribution of cholesterol in thin sections and freeze-fracture replicas. Filipin-sterol complexes were not homogeneously distributed throughout the spermatozoon plasma membrane. They were more abundant on the P-fracture face of the membrane lining the nuclear region. The results obtained show that Culex spermatozoon differs from those of other species in that its plasma membrane exhibits only a membrane domain, the zipper-line, localized in the postacrosomal region.


Lymphocyte calmodulin and its participation in the stimulation of T lymphocytes by mitogenic lectins. Calmodulin was purified from human tonsillar lymphocytes utilizing calcium-dependent binding of calmodulin to fluphenazine-Sepharose. The molecular weight and phosphodiesterase activation of the lymphocyte calmodulin were very similar to those of purified bovine brain calmodulin. Trifluoperazine (TFP), a calmodulin inhibitor, suppressed lymphocyte stimulation as assessed by 3H-thymidine incorporation into DNA of lectin-stimulated lymphocytes. TFP had no effect on the early 45Ca2+ uptake induced by mitogenic lectins, although this latter was inhibited by verapamil which also suppressed the 3H-thymidine incorporation. The results are in keeping with the interpretation that the inhibition of T cell stimulation by TFP was not due to suppression of Ca2+ uptake, but due to inactivation of Ca(2+)-calmodulin complex which might be formed subsequent to Ca2+ entry into the cell.


Differential temperature sensitivity of cultured cells from cartilaginous or bone origin. The effects of long-term exposure to hyperthermia were studied on several cell cultures of cartilaginous or bone origin after a 4-day treatment at 40 degrees C. Chondrocytes proliferation, as well as mitochondrial activity were not modified by these culture conditions (40 degrees C) but protein content and cell volume were increased. In contrast, the proliferative capacity of osteoblasts, MC3T3.E1 a and ROS 17/2.8 was decreased and their protein content, cell volume and mitochondrial activity were increased. Chondrocytes appeared to be thermoresistant, and osteoblastic cells thermosensitive. Furthermore, temperature sensitivity was greater for the continuous established osteoblastic cell line MC3T3.E1 and for the cancerous established osteoblastic cell line ROS 17/2.8 than for chondrocytes.


Initiation and stability of reentry in two coupled excitable fibers. Reentry in the heart is the repeated excitation of the same tissue by a single excitation wave; it is responsible for several types of cardiac arrhythmia. The simplest model which permits the phenomenon of reentry is two laterally coupled excitable fibers; in this paper we examine such a model in order to establish a basis for the understanding of the fundamental physical processes underlying the process of reentry. Two versions of the FitzHugh-Nagumo equations are used to develop complementary numerical and analytical results for the coupled fiber model. On the basis of numerical studies, regions of qualitatively different behaviour are mapped in the parameter space of excitation threshold and coupling strength between the fibers, and the effect of the rate of recovery is explored. Some of these regions are also obtained analytically, in good agreement with the numerical results. Finally, the results are discussed in the light of recent work on the role of the anisotropy of cardiac tissue in the initiation of reentrant activity in the heart.


Modelling the electrotonic structure of starburst amacrine cells in the rabbit retina: a functional interpretation of dendritic morphology. A detailed morphometric analysis of a Lucifer yellow-filled Cb amacrine cell was undertaken to provide raw data for the construction of a neuronal cable model. The cable model was employed to determine whether distal input-output regions of dendrites were electrically isolated from the soma and each other. Calculations of steady state electrotonic current spread suggested reasonable electrical communication between cell body and dendrites. In particular, the centripetal voltage attenuation revealed that a synaptic signal introduced at the distal end of the equivalent dendrite could spread passively along the dendrite and reach the soma with little loss in amplitude. A functional interpretation of this result could favour a postsynaptic rather than a presynaptic scheme for the operation of directional selectivity in the rabbit retina. On the other hand, dendrites of starburst amacrine cells process information electrotonically with a bias towards the centrifugal direction and for a restricted range of membrane resistance values the voltage attenuation in the centripetal direction suggests that the action of these dendrites can be confined locally. A functional interpretation of this result favours a presynaptic version of Vaney's cotransmission model which attempts to explain how the neural network of starburst amacrine cells might account for directionally selective responses observed in the rabbit retina.


Flow-induced deformation from pressurized cavities in absorbing porous tissues. The behaviour of a cavity during an injection of fluid into biological tissue is considered. High cavity pressure drives fluid into the neighbouring tissue where it is absorbed by capillaries and lymphatics. The tissue is modelled as a nonlinear deformable porous medium with the injected fluid absorbed by the tissue at a rate proportional to the local pressure. A model with a spherical cavity in an infinite medium is used to find the pressure and displacement of the tissue as a function of time and radial distance. Analytical and numerical solutions for a step change in cavity pressure show that the flow induces a radial compression in the medium together with an annular expansion, the net result being an overall expansion of the medium. Thus any flow induced deformation of the material will aid in the absorption of fluid.


The role of cytokines in graft-versus-host reactions and disease. CD4+ and CD8+ T lymphocytes are responsible for the initiation of graft-versus-host disease (GVHD) which limits the efficacy of human allogeneic bone marrow transplantation. Activated T cells are a rich source of cytokines in vitro, and many of these same molecules are also found in graft-versus-host reactions (GVHR) and GVHD. However, T cells are not the only source of these soluble mediators. Cytokines control the actions of many cell types both in vitro and in vivo, so these soluble factors can easily influence the actions of both the engrafted hematopoietic cells and the host's cells. The complex manner in which cells and cytokines interact in GVHR and GVHD is reviewed here. We speculate that cytokine cascades play major roles in many aspects of GVHR and GVHD.


The role of conditioning regimens in homing of transplanted hemopoietic cells. Evidence is reviewed to indicate that the conditioning regimens given to prepare the patient for bone marrow transplantation have two unintended but desirable consequences: (a) they eliminate endogenous progenitor cells that may otherwise be in competition with grafted hemopoietic progenitor cells, leaving all homing sites open for transplanted cells, and (b) they induce membrane alterations in marrow endothelium, thus effectively removing the endothelial barrier which otherwise the grafted cells must negotiate to enter the hemopoietic compartment of the marrow. Hence the conditioning regimens may also facilitate the homing of transplanted cells.


Proliferation rates in epidermis of patients with graft-versus-host disease, non-specific inflammation and normal skin. There is evidence that epidermal stem cells or their early progeny may be the targets in graft-versus-host disease (GVHD). The early progeny should be a proliferating group of daughter cells more concentrated in areas just above or adjacent to stem cell regions. We have, therefore, compared proliferative rates of keratinocytes within relevant subregions of the epidermis using the AgNOR stain on biopsies from patients with GVHD, non-specific inflammatory infiltrates (NSI) and normal skin. We concurrently evaluated T cell infiltration using immunohistology on paraffin-embedded tissue with UCHL-1 antibody. Fifty-one bone marrow transplant patients were evaluated in each of three temporal groups (days 7-20 post-transplant, days 20-40 post-transplant, and days 80-100 post-transplant). In each of these groups, 5-9 patients with a histological diagnosis of GVHD were compared to similar numbers of patients with a histological diagnosis of non-specific dermatitis.(ABSTRACT TRUNCATED AT 250 WORDS)


Autologous bone marrow transplantation in 22 adult patients with lymphoblastic lymphoma responsive to conventional dose chemotherapy. In order to clarify the role of autologous bone marrow transplantation in adult lymphoblastic lymphoma (LBL) patients we have conducted a retrospective multi-institutional Spanish survey. Twenty-two adult patients, 20 males, two females, age 14-52 years (median 25.5) with LBL were treated with high-dose chemoradiotherapy and autologous bone marrow support. Fourteen cases were transplanted in first complete remission (CR1) and eight with other chemosensitive status (four later CR, two partial remissions and two sensitive relapses). From the 14 cases transplanted in CR1, four had previous bone marrow involvement and one meningeal infiltration; eight cases were Ann-Arbor stage IV and fulfilled accepted high-risk criteria for relapse. The conditioning regimen consisted of cyclophosphamide (60 mg/kg x 2) and total body irradiation (9-12 Gy) in 16 cases and high dose chemotherapy in six. The procedure-related mortality was 9% (7% in CR1 patients). The actuarial 2-year overall survival for CR1 patients was 85% at a median follow-up of 19 months. Disease-free survival (DFS) was 77%. In patients with less favourable disease status the 2-year overall and DFS were 73% and 50% respectively. In this study the DFS in CR1 patients was not influenced by bone marrow involvement or high-risk criteria predictive for relapse. These results support the effectiveness of this procedure, mainly for patients in CR1.


Marrow transplantation for treatment of multisystem progressive Langerhans cell histiocytosis. Langerhans cell histiocytosis (LCH) embraces disorders characterized by the presence of marrow-derived abnormal Langerhans cells. A small number of patients with multisystem disease, vital organ dysfunction and rapid disease progression are considered to have a poor prognosis despite various treatments. Antiproliferative and immunosuppressive therapy, in combination with marrow transplantation, could be the appropriate treatment for these poor-prognostic patients. Four patients with progressive LCH were treated with high dose chemotherapy and fractionated total body irradiation followed by either an allogeneic (n = 2) or autologous (n = 2) marrow graft. Two of them are alive and have been disease free for almost 2 and 4 years after marrow grafting, respectively. One of the two survivors is the recipient of an allogeneic and the other of an autologous marrow graft. Two patients died, one of intrapulmonary hemorrhage 14 days after transplantation with active disease at autopsy, and the other of relapse of the original disease 355 days after marrow grafting. For patients with multisystem progressive LCH, allogeneic and autologous marrow transplantation may offer an opportunity for long-term remission and disease-free survival.


Influence of the conditioning regimen on erythrocyte chimerism, graft-versus-host disease and relapse after allogeneic transplantation with lymphocyte depleted marrow. Three different conditioning regimens were applied to 144 patients undergoing allogeneic bone marrow transplantation (BMT) with HLA identical sibling marrow, depleted of lymphocytes by counterflow centrifugation. All regimens consisted of cyclophosphamide and fractionated total body irradiation (TBI). In 49 patients treated with regimen A the total TBI dose was 9 Gy. In regimen B the dose rate of TBI was increased and anthracyclines were added (n = 65). Thirty patients received regimen C with a total TBI dose of 12 Gy but no anthracyclines. The different conditioning regimens did not influence the percentage of patients with detectable recipient CFU-GM prior to infusion of donor marrow. The incidences of mixed erythrocyte chimerism at 6 months after BMT were 73, 33 and 20% for regimens A, B and C respectively. The conditioning regimen influenced significantly mixed erythrocyte chimerism from 6 to 24 months after BMT. Both age and the conditioning regimen influenced significantly the incidence of acute graft-versus-host disease (GVHD) (p = 0.017 and 0.0001 respectively). Acute GVHD greater than or equal to I occurred in 15, 29 and 77% of the patients treated with regimens A, B and C respectively. The incidence of acute and chronic GVHD was significantly higher in complete donor chimeras than in mixed chimeras (p less than 0.001 and p less than 0.01). The probability of relapse was 43% in 32 and 18% in 43 good risk patients treated with regimens A and B respectively (p = 0.07). Longer follow-up is needed to draw conclusions about relapse in regimen C.(ABSTRACT TRUNCATED AT 250 WORDS)


Allogeneic bone marrow transplantation for Fanconi anemia. Five patients (age range 7-14 years) received allogeneic bone marrow transplantation (BMT) for Fanconi anemia (FA). All patients showed progressive pancytopenia associated with congenital malformations. Diagnosis was confirmed by studies of cellular hypersensitivity to the clastogenic effect of the DNA crosslinking agent diepoxybutane. The conditioning regimen consisted of low dose cyclophosphamide (5 mg/kg x 4) and fractionated total body irradiation (167 cGy x 3). For graft-versus-host disease prophylaxis one patient was given cyclosporin alone while the remaining four patients received a combination of cyclosporin and two doses of methotrexate. Marrow was given unmanipulated from HLA-identical siblings. All patients are alive 18-67 months after grafting with Karnofsky scores of 100% and normal hemopoiesis of donor origin. Modifications in transplant protocols such as those here described have resulted in a decreased risk of severe transplant-related complications. These results confirm that BMT is a curative therapy in FA patients and should be considered as a first choice treatment if an HLA-identical donor is available.


Pulmonary toxicity associated with high dose chemotherapy in the treatment of solid tumors with autologous marrow transplant: an analysis of four chemotherapy regimens. We retrospectively reviewed the pulmonary toxicity of six high dose chemotherapy protocols using four chemotherapy regimens in the treatment of solid tumors. All protocols used either high dose cyclophosphamide or ifosfamide in combination with one to three additional chemotherapeutic agents. In each protocol autologous bone marrow was reinfused post chemotherapy to shorten the period of severe myelosuppression. Of 178 patients there were 20 cases of fatal or life-threatening pulmonary toxicity including nine cases of pneumonia, nine cases of interstitial pneumonitis and two cases of pulmonary hemorrhage. Pulmonary function tests revealed modest changes in FEV1 and DLCO in the majority of patients, although 24 patients had more dramatic changes in DLCO suggesting interstitial damage. Significant decrements in FEV1 were seen in the BCNU containing regimen. Statistically significant or nearly significant decreases in DLCO were seen after all cyclophosphamide containing regimens. A regimen containing ifosfamide, carboplatin, and etoposide had minimal associated pulmonary toxicity.


Pancreatitis associated with bone marrow transplantation in children. Vomiting, abdominal pain and distension, common findings in children who receive bone marrow transplants (BMT), are usually attributed to chemo-irradiation and mucositis, universally found in these patients. We report seven children, 3.5% of BMT patients at our institutions, with these symptoms who were found to have mild to severe pancreatitis during conditioning for or after receiving BMT. All patients were receiving drugs known to cause pancreatitis, such as adrenocorticosteroids and sulfonamides as well as numerous putative pancreatotoxins such as cyclosporin A and cytosine arabinoside. Five of the seven patients had suffered from graft-versus-host disease. In patients who have received BMT, upper gastrointestinal symptoms should not be attributed to mucositis or chemo-irradiation without first testing for pancreatitis.


Serum erythropoietin levels and blood component therapy after autologous bone marrow transplantation: implications for erythropoietin therapy in this setting. To determine the potential impact of recombinant human erythropoietin (EPO) therapy in patients undergoing autologous bone marrow transplantation (BMT) and colony-stimulating factor therapy, we assayed endogenous serum EPO levels and noted blood transfusion requirements in relapsed non-Hodgkin's lymphoma patients treated with high-dose chemo-radiation therapy and autologous BMT. Hematocrit and reticulocyte counts were determined daily, and hematocrit was maintained in the 25-30% range by transfusion according to criteria established by our hospital transfusion committee. EPO levels were measured by radioimmunoassay and were determined at baseline, throughout therapy, and 2 and 3 months after BMT. Serum EPO levels increased more than 25-fold above baseline in most subjects after initiating chemoradiation therapy. No correlation was noted between serum EPO and hematocrit, reticulocyte count or serum creatinine. Total red blood cell units transfused ranged from 4 to 15 (mean 7.7). Mean total donor exposures (red blood cell plus platelet units transfused) were 83.6 units (range 16-175). Serum EPO levels increased early in the course of preparation for autologous BMT and remained elevated for at least 2-3 weeks thereafter although at a lower level. Red blood cell transfusions were required despite very high EPO levels after BMT. Red cell transfusions, moreover, accounted for only 9.2% (69 of 746) of total donor exposures and only 5.8% (42 of 746) of donor exposures during the interval when pharmacologic doses of erythropoietin might be of benefit. In contrast to the potential benefit of colony-stimulating factors such as G-CSF and GM-CSF in BMT, our study suggests limited value for erythropoietin therapy in this setting.


Acute graft-versus-host disease following unrelated donor marrow transplantation: failure of conventional therapy. The response to therapy of acute graft-versus-host disease (GVHD) is uncertain in recipients of unrelated donor (URD) bone marrow transplant (BMT). We analysed the outcome of treatment in 42 patients with moderate/severe acute GVHD. Initial therapy consisted of prednisone 60 mg/m2 orally daily for 7 days (n = 42), followed by anti-thymocyte globulin (ATG) 15 mg/kg i.v. twice daily for 8-10 doses after prednisone failure (n = 22). A clinical Stage Score for acute GVHD was determined initially and after 7, 14, 21, and 28 days of prednisone or ATG. Treatment failure represented worsening score after 7 days, involvement of a new organ or failure to improve after 14-28 days. Prednisone treatment led to 10 of 41 (24%) patients improving, while secondary therapy with ATG led to four of 21 (19%) improving. Of 42 patients treated, only nine (21%) achieved a complete and continuing response of acute GVHD by day +100. Neither age, diagnosis, recipient/donor gender status, histocompatibility nor GVHD prophylaxis regimen was associated with more frequent responses. Response to GVHD therapy was significantly correlated with survival at 100 days and 1 year post-BMT. We conclude that prednisone and ATG used for treatment of acute GVHD following URD BMT are associated with a high failure rate and that more aggressive therapy is warranted in these patients.


A modified harvest technique for cord blood hematopoietic stem cells. Human umbilical cord blood is an excellent source of hematopoietic stem cells for research and bone marrow transplantation. We have developed a modified technique which effectively flushes and collects placental derived stem cells. Catheters were inserted into the umbilical artery and vein. Using this sterile and closed system technique, 10 saline diluted cord blood harvests had hematocrits of 16% to 25%, total mononuclear cell counts of 1.17 to 34.4 x 10(8) and volumes of 105-245 ml. The averaged extrapolated colony forming units from five cord blood harvests were as follows: CFU-GM 5.2 x 10(5) (9.96 x 10(4) to 1.24 x 10(6)), BFU-E 8.82 x 10(5) (2.40 x 10(4) to 1.61 x 10(6)) and CFU-GEMM 1.37 x 10(5) (7.33 x 10(3) to 3.18 x 10(5)). This quick technique avoids needle exposure and collects a significant volume of stem cells.


Etoposide, cyclophosphamide and fractionated total body irradiation as a preparative regimen for marrow transplantation in patients with advanced hematological malignancies: a phase I study. Thirty-seven patients with advanced hematologic malignancy were entered into a phase I study designed to define a maximum tolerable dose (MTD) of etoposide (VP-16) and cyclophosphamide (CY) combined with 12 Gy fractionated total body irradiation (TBI) as preparation for marrow transplantation from an HLA-identical sibling (n = 13) or with cryopreserved autologous marrow (n = 24). Dose levels ranged from 36 mg/kg of VP-16 combined with 67 mg/kg of CY to 52 mg/kg of VP-16 combined with 103 mg/kg of CY followed by 12 Gy TBI. The MTD for allogeneic marrow recipients was 36 mg/kg of VP-16 + 52 mg/kg of CY followed by 12 Gy TBI and for autologous marrow recipients 44 mg/kg of VP-16 + 103 mg/kg CY followed by 12 Gy TBI. Pulmonary and liver toxicities were dose limiting. All of 31 evaluable patients transplanted in relapse achieved a complete remission. However, in all but three of these patients the disease relapsed 28-899 (median 110) days post-transplant. Currently, six of 24 autologous marrow recipients are surviving, three in remission 256, 340 and 764 days post-transplant. None of 12 allogeneic marrow recipients have survived. In conclusion, a preparative regimen combining 44 mg/kg of VP-16 + 103 mg/kg CY followed by 12 Gy TBI is well tolerated by autologous marrow recipients and 36 mg/kg VP-16 + 67 mg/kg CY followed by 12 Gy TBI is well tolerated by allogeneic marrow recipients.(ABSTRACT TRUNCATED AT 250 WORDS)


Successful pregnancy after total body irradiation and bone marrow transplantation for acute leukaemia. We report successful pregnancies in two young women (aged 24 and 20 years) following allogeneic bone marrow transplantation (BMT) for acute non-lymphoblastic leukaemia. Conditioning therapy consisted of cyclophosphamide (120 mg/kg) and total body irradiation (TBI, 12 Gy) in 2 Gy fractions once daily for 6 days or twice daily for 3 days. Graft-versus-host disease prophylaxis was with methotrexate alone. Both women were amenorrhoeic after BMT and gonadal testing indicated hypergonadotrophic hypogonadism. Both women had normal pregnancies (2 years and 5 years after BMT) resulting in normal healthy infants. Previously successful pregnancy has been reported after TBI in three women in whom the TBI dose was less than 8 Gy. Our cases illustrate that normal outcome of pregnancy is possible at even higher doses of TBI.


A comparison of ultrasound exposure in therapy and pulsed Doppler fields. A detailed comparison of the ultrasound exposure in water from a therapeutic beam and a pulsed Doppler beam was carried out. A significant overlap in acoustic power was found between therapy intensity levels used clinically and the upper end of the diagnostic range, between approximately 100 mW and 200 mW. In addition, pulse pressure amplitudes in the range 0.5-1.0 MPa were measured close to the transducer on both units. It is common to use physiotherapy equipment at pulse average intensities of 0.5 W/cm2 or less, and at these levels exposures of similar magnitude may be obtained with beams currently defined as therapeutic and those available from pulsed Doppler equipment.


Routine radiographer screening for fetal abnormalities by ultrasound in an unselected low risk population. A screening programme for fetal abnormalities began at The Hillingdon Hospital in July 1986. Second trimester ultrasound scans are performed by radiographers. A combined prospective and retrospective study of the ultrasound findings and outcome in all pregnancies delivered in 1989-1990 was undertaken. 6412 babies were born during this period, of whom 6183 (96%) were examined by ultrasound in the second trimester; 29 pregnancies were terminated for fetal abnormality. Of the 89 fetuses who were abnormal at birth or at induced termination of the pregnancy (1.4%), 84 were scanned in the second trimester. In 51 cases the abnormality was detected before 22 weeks gestation (sensitivity, 60.7%). 56 of these 84 abnormal fetuses scanned had potentially lethal or major handicapping abnormalities of which 41 were detected by ultrasound before 22 weeks gestation (sensitivity, 73%). There was one false positive diagnosis of abnormality which did not affect outcome. 6352 babies were normal at delivery or on discharge from hospital (specificity, 99.98%).


A study of the relationship between routine ultrasound quality assurance parameters and subjective operator image assessment. Comparisons have been made between a number of routine quality assurance (QA) measurements and a number of related subjective aspects of image quality. These comparisons between lateral resolution and perceived definition, dynamic range and subjective grey-level assessment, and slice thickness and overall image quality appraisal showed very poor correlation. We deduce from this that other QA parameters are needed to quantify good image quality as defined by the operators.


A role for ultrasound in limb lengthening. The distraction gap at eight sites in six patients undergoing limb lengthening was monitored by both standard radiographs and ultrasound. Using the measurements obtained we were able to assess the rate and quality of callus formation at the distraction site, something that radiographs alone do not enable one to do. We suggest that ultrasound has a valuable role to play in guaging the optimum rate of lengthening for a particular patient and commend it as a technique to complement radiographs in the follow-up of the increasing numbers of patients undergoing distraction osteogenesis.


An audit of ultrasound diagnosis of gallbladder calculi. Many series suggest that ultrasound is an accurate method for demonstrating cholelithiasis. However, these series were often prospective and the examinations performed by experienced sonographers. This audit addresses whether the accuracy is maintained in daily practice. We reviewed the ultrasound scans of 128 patients who underwent cholecystectomy for cholelithiasis and compared the findings. The operative and ultrasound findings were at variance in eight of the 128 patients (6.2%). Five were reported as having gallstones on ultrasound but none were found at cholecystectomy, a false positive rate for ultrasound of 3.9%. Three had abnormal gallbladders with no gallstones on ultrasound but gallstones were found at operation, a false negative rate of 2.3%. To avoid false positive diagnoses, suboptimal examinations should be repeated and the scan should be repeated immediately pre-operatively if only small calculi are seen. Alternative imaging should be performed if necessary, either cholescintigraphy in the acute situation or elective oral cholecystography. Some false negative examinations may be avoided by performing repeat examinations if the gallbladder is thick-walled and tender. With these provisos we conclude that ultrasound correctly diagnoses cholelithiasis in daily practice.


The ultrasound appearances of intramural bowel gas: the bright ring appearance and the effervescent bowel. A report of three cases. Bowel necrosis is a serious condition with a very high mortality rate. The earlier the diagnosis is made, the better chance for survival. Ultrasound is a primary imaging technique in the diagnosis of the acute abdomen. Three cases are presented in which bowel wall gas could be demonstrated on ultrasound. In all cases there was computed tomographic and either operative or post-mortem correlation. In each case there was a "bright ring" appearance of the affected bowel when the ultrasound examination was carried out in transverse section. In one case, small bubbles were seen arising from the deep surface of the bowel.


The value of ultrasound in the assessment of bone. The impenetrability of bone to ultrasound can be turned to advantage and the effects of bony lesions on the adjacent soft tissues can be analysed. Fractures, bone destruction, inflammation, distraction of bone and new bone formation can be diagnosed.


Conventional 15 G needle technique for renal biopsy compared with ultrasound-guided spring-loaded 18 G needle biopsy. Two techniques for percutaneous renal biopsy were retrospectively reviewed to assess their relative safety and efficacy. Ultrasound localization of the kidney by a radiologists, with subsequent biopsy performed by a renal physician using a hand-held 15 G cutting needle (Tru-Cut), was compared with biopsy performed by a radiologist using an 18 G cutting needle with a spring-loaded biopsy device (Biopty) and real-time ultrasound guidance. The smaller needle with real-time ultrasound is more reliable at retrieving an adequate specimen for histological examination (93%) than the "conventional" technique (79%). Fewer complications occurred in the Biopty group although the difference did not reach statistical significance. The average length of stay in hospital was significantly shorter for elective biopsies with the Biopty device (1.80 compared with 2.93 nights, p less than 0.01). We recommend the use of the Biopty device with an 18 G needle and real-time ultrasound guidance as the method of choice for percutaneous renal biopsy.


Opposite effects of prefrontal cortex and nucleus accumbens infusions of flupenthixol on stimulant-induced locomotion and brain stimulation reward. Ventral tegmental area (VTA) stimulation produced conditioned place preferences for stimulation-paired environments the magnitudes of which were dose-dependently reduced by systemic application of the dopamine antagonist, haloperidol (0.0, 0.15, 0.3 mg/kg). Bilateral microinjections of cis-flupenthixol (FLU) into the nucleus accumbens (0.0, 1.0, 5.0 or 10.0 micrograms) also resulted in reductions in the size of stimulation-induced place preferences as well as reductions in the magnitude of the hyperlocomotor response to 1.5 mg/kg (s.c.) D-amphetamine. Comparable microinjections of FLU into the medial prefrontal cortex (PFC) produced diametrically opposite effects: the size of VTA stimulation-induced place preferences was either unaffected (1.0 and 5.0 microgram groups) or slightly increased (10 micrograms group) and amphetamine-stimulated hyperlocomotion was dose-dependently potentiated. These behavioral findings suggest a dopamine-mediated modulatory role for the PFC over reward relevant elements within the nucleus accumbens.


Expression, but failing maturation of procholecystokinin in cerebellum. The cerebellum is the only region of the central nervous system which has been found to be devoid of cholecystokinin (CCK). The assays used, however, have been directed against the alpha-amidated C-terminus of fully processed CCK peptides. Using Northern blot analysis and a library of radioimmunoassays specific for different sequences of proCCK in combination with chromatography and enzyme cleavage, we have now examined the expression and processing of proCCK in fetal, neonatal and adult cerebellar tissue from man, pig and rat. In rat cerebellum CCK mRNA was present already in the fetal state. Two weeks after birth the concentrations declined. Also proCCK was found in significant concentrations in the fetal human and rat cerebellum (approximately 20 pmol/g); but already before birth the expression began to decrease towards low concentrations in adults. The adult porcine cerebellum contained 3.2 pmol proCCK and glycine-extended processing intermediates per gram (range less than 0.1-10.4 pmol/g), and 0.8 pmol carboxyamidated CCK per gram (range 0.1-4.1 pmol/g) varying in size from CCK-58 to CCK-5. For comparison, the adult porcine cerebral cortex contained 757 pmol carboxyamidated CCK/g, 20 pmol glycine-extended CCK/g and no proCCK. We conclude that cerebellum expresses proCCK with the highest level of expression in fetal life. In comparison with other regions of the brain, the maturation to transmitter-active, carboxyamidated CCK peptides is, however, attenuated in both fetal and adult cerebellar tissue.


Tonic descending modulation of spinal neurons with renal input. Experiments were conducted to determine the influence of tonically active descending pathways on thoracolumbar spinal neurons that respond to renal nerve stimulation in anesthetized cats. We examined the effect of reversible blockade of spinal conduction on spontaneous activity, responses to renal nerve stimulation and responses to somatic stimuli of 71 spinal neurons. Mid-thoracic cold block resulted in enhanced responses (tonically inhibited neurons), reduced responses (tonically excited neurons), or did not affect neuronal responses. The spontaneous activity of 47 of 69 neurons (68%) increased from 7.3 +/- 2.0 spikes/s before cooling to 23.3 +/- 4.5 spikes/s during cooling. Activity of 8 neurons (12%) decreased while 14 (20%) had no change in activity. Cooling increased the responses of 51 of 71 neurons (72%) to renal nerve stimulation. Renal nerve stimulation evoked a two-fold increase in both short latency (early) and long latency (late) responses. Four neurons had a late response which was revealed by cold block. Cooling decreased responses of 8 of 71 neurons (11%) and 9 neurons (13%) were not affected. Cooling increased the early responses but decreased the late responses of 3 of 71 neurons (4%). All neurons had somatic receptive fields and 33 of 56 exhibited increased responses to somatic stimulation during cooling. In addition, receptive field sizes of 26 neurons increased. Four neurons had a decrease and 25 neurons had no change in receptive field size during cooling. These data indicate that tonically active descending pathways modulate the activity of most spinal neurons with renal input and the major effect of these pathways is inhibitory. This influence may be important in the modulation of spinal circuits that participate in reflexes evoked by renal afferent fibers.


Selective inhibition of glial cell metabolism in vivo by fluorocitrate. The effect of fluorocitrate on glial and neuronal amino acid metabolism was studied. One nmol of fluorocitrate administered intrastriatally in the rat caused a 95% reduction of glutamine formation from [14C]acetate, a substrate which enters the glial cells selectively. The metabolism of [14C]glucose which enters neurons, was unaffected by fluorocitrate treatment except for the glutamine formation. This is evidence that fluorocitrate is a selective inhibitor of the glial Krebs' cycle. [14C]Citrate and 2-oxoglutarate labelled amino acids in a manner similar to [14C]acetate, which shows that these substrates are taken up and metabolized by glial cells. Differences in the labelling of gamma-aminobutyric acid (GABA) from [14C]acetate and citrate suggest that astrocytes associated with GABAergic and glutamatergic nerve terminals may differ in their preference for amino acid precursors.


Estrogen fails to reduce tuberoinfundibular dopaminergic neuronal activity and to cause a prolactin surge in lactating, ovariectomized rats. The activity of tuberoinfundibular dopaminergic (TIDA) neurons was estimated by measuring the concentrations of dihydroxyphenylacetic acid (DOPAC) in the median eminence of ovariectomized lactating rats, ovariectomized non-lactating female rats and castrated male rats in correlation with the estrogen-induced prolactin (PRL) surge. PRL surge with a peak at 17.00 h was observed in ovariectomized non-lactating rats but not in the other groups. DOPAC levels were significantly lower at 17.00 h than at 11.00 h in non-lactating female rats, but there was no change of DOPAC levels in lactating rats and male rats. These results indicate that TIDA neuronal activity is inhibited at the time of estrogen-induced PRL surge and that the TIDA neurons may alter their function to result in refractoriness of PRL response to estrogen in the lactating rat.


Lateral dominance and motor unit firing behavior. Twelve subjects were classified as left-handed (LH) or right-handed (RH) using Annett's hand dominance classification. Motor unit recordings were obtained from the first dorsal interosseous (FDI) muscle of each hand using a quadrifilar needle electrode. Firing occurrences of individual motor units were then identified and the firing rates of all motor units recorded during the contraction were cross-correlated. The results demonstrated significantly greater firing rate cross-correlation scores in the dominant hand than in the non-dominant hand for both LH and RH subjects. This association between hand dominance and the common drive of motor unit firing rates lends credence to the idea that one or more CNS sites may influence conjoint motor unit firing behavior.


A cocaine analog and a GBR analog label the same protein in rat striatal membranes. Because some evidence suggests that cocaine and GBR12935 bind to different sites, we utilized photoaffinity probes from both classes of compounds to see if they label the same protein. [125I]RTI-82 a cocaine analog, and [125I]DEEP, a GBR analog, labeled protein(s) showing the same molecular weight, a similar pharmacological profile and a similar sensitivity to neuraminidase.


Ectopic axonal firing in an epileptic cortical focus is not triggered by thalamocortical volleys during the interictal stage. The triggering of ectopic action potentials (APs) at axon terminals in a chronic epileptic cobalt focus was investigated in thalamocortical (TC) neurons of rats under urethane anesthesia. TC cells which were in register with an active epileptic aggregate discharged bursts of 2-11 APs. According to the rules of the collision test, we ascertained that bursts contained APs of ectopic and/or somatic origin. During a transient blockage of TC orthodromic discharges produced by raising the extracellular concentration of Mg2+, ectopic bursts which were in close time relationship with the focal interictal electrocorticographic spikes persisted. These results demonstrate (i) the antidromic nature of identified ectopic APs and (ii) that, during the interictal stage, such axonal APs were not a consequence of TC discharges. The possible mechanisms for the triggering of ectopic axonal APs in the chronic cobalt focus are discussed.


Cellular variations in estrogen receptor mRNA translation in the developing brain: evidence from combined [125I]estrogen autoradiography and non-isotopic in situ hybridization histochemistry. The spatial distribution of cells in the adult rodent forebrain which express estrogen receptor mRNA, as shown by in situ hybridization histochemistry with isotopically-labeled probes, has been reported to overlap with regions that are known targets of estrogen and which bind estrogen. The extent to which detection of estrogen receptor mRNA within developing forebrain neurons of the postnatal day 10-12 female rat is accompanied by translation into estrogen binding sites was investigated by combining [125I]estrogen autoradiography with non-isotopic (digoxigenin) in situ hybridization, using a 48-base oligodeoxyribonucleotide probe encoding a sequence of the estrogen-binding domain of rat uterine estrogen receptor cDNA. Estrogen receptor mRNA and estrogen binding sites appeared to be restricted to neurons. No mRNA or binding was seen in ependymal cells. Cells expressing estrogen receptor mRNA were widely distributed in the developing rat forebrain and were found in brain regions generally corresponding to those previously shown in the adult, with the addition of some regions not previously described, such as the medial habenula and dorsal endopiriform nucleus. Although there was widespread overlapping of estrogen receptor mRNA expression with known estrogen binding sites, there were regional and cellular variations in the extent of receptor mRNA translation. This pattern was true for developing forebrain regions previously defined as estrogen receptor-containing (hypothalamus, preoptic area, medial and lateral septum, vertical and horizontal nuclei of the diagonal band, cerebral cortex, hippocampus and amygdala) as well as for regions heretofore not considered estrogen targets (the thalamus, dorsal endopiriform nucleus, claustrum, ventral pallidum/substantia innominata and the basal nucleus of Meynert) or characterized as estrogen-responsive in the adult without previously documented estrogen binding [caudate-putamen (striatum)]. While estrogen binding and receptor mRNA expression always co-localized, neurons expressing estrogen receptor mRNA did not always exhibit ligand binding and there was no clear-cut relationship between the intensity of the hybridization signal and estrogen binding. Little, however, is known about translational control of estrogen receptor expression in the brain. Localization of estrogen binding sites to regions not generally considered targets of estrogen would appear to reflect the greater sensitivity of the iodinated ligand than the tritiated estrogens more commonly used for autoradiography. Non-isotopic in situ hybridization histochemistry combined with [125I]estrogen autoradiography represents a very powerful tool with which to study regulation of estrogen receptor gene expression at the single cell level with an exceptional degree of cellular and anatomical resolution.(ABSTRACT TRUNCATED AT 400 WORDS)


Recovery in hemiparkinsonian rats following intrastriatal implantation of activated leukocytes. Behavioral improvement has been seen in hemiparkinsonian animals after surgical lesions of the denervated caudate nucleus. This study was designed to investigate the role of inflammatory cells in injury-induced recovery. A hemiparkinsonian syndrome was induced in rats by unilateral injections of 6-hydroxydopamine into the pars compacta of the substantia nigra. Phytohemagglutinin-stimulated rat peritoneal cells, predominantly T cells and macrophages, were stereotactically implanted in the lesioned caudate-putamen, and amphetamine-induced turning was used to assess recovery. Animals receiving implants of activated peritoneal cells showed a 47% decrease in amphetamine-induced turning 8 weeks after implantation, which was not seen in control or sham-operated animals. Immunocytochemistry revealed increased tyrosine hydroxylase reactive fibers in the leukocyte-implanted striatum. We conclude that implantation of activated leukocytes promotes functional recovery in hemiparkinsonian rats.


Messenger RNAs encoding glutamate-decarboxylases are differentially affected by nigrostriatal lesions in subpopulations of striatal neurons. Dopaminergic nigrostriatal neurons constitute one of the major inputs to the striatum, and play a role in the regulation of gamma-aminobutyric acid (GABA) and glutamic acid decarboxylase (GAD), the GABA-synthesizing enzyme, in striatal neurons. The effect of nigrostriatal lesions on the level of expression of messenger RNAs encoding two distinct isoforms of glutamate decarboxylase was examined at the single cell level with in situ hybridization histochemistry. Rats received a unilateral injection of the neurotoxin 6-hydroxydopamine in the substantia nigra and were sacrificed 2 or 3 weeks later. Sections of the striatum were processed for in situ hybridization histochemistry with radiolabeled RNA probes selective for mRNAs encoding glutamate decarboxylase with molecular weights of 65,000 and 67,000, respectively. In addition, immunohistochemistry with a monospecific antibody for the latter glutamate decarboxylase isoform was performed. In agreement with previous reports, we observed increased labeling for the messenger RNA encoding glutamate decarboxylase (M(r) 67,000) in a population of medium-sized striatal efferent neurons normally expressing low levels of this messenger RNA. We now show that this effect occurred in two striatal compartments, the striosomes and the extrastriosomal matrix, and was accompanied by increased immunostaining for the corresponding protein with a monospecific antibody. In contrast, labeling for messenger RNA encoding GAD (M(r) 67,000) was decreased in a population of medium-sized neurons normally expressing high levels of this messenger RNA and corresponding to GABAergic interneurons. Labeling for messenger RNA encoding glutamate decarboxylase (M(r) 65,000) was not modified in the dopamine-depleted striatum. The results show that dopamine depletion differentially affects gene expression for different isoforms of glutamate decarboxylase in distinct subpopulations of striatal neurons in rat.


Halothane-induced hyperpolarization and depression of postsynaptic potentials of guinea pig thalamic neurons in vitro. Intralaminar thalamic nuclei have been considered to be a component of the non-specific sensory system which is involved in physiological functions related to consciousness and pain sensation. The effect of halothane on membrane potentials and synaptic properties of neurons of the parafascicular (Pf) nucleus in guinea pig brain slices was investigated using intracellular recording methods. Halothane at concentrations of 0.4-1.0 mM, which are in the range of clinical concentrations, produced hyperpolarizations of 2-8 mV in approximately 50% of the cells. The halothane-induced hyperpolarization was nullified at a membrane potential close to the K+ equilibrium potential. The amplitude of the hyperpolarization was dependent on the external K+ concentration, and was decreased by either Ba2+, or 4-aminopyridine, or intracellular injection of Cs+. All these results indicate that the hyperpolarization was due to an increase in K+ conductance. Halothane at clinical concentrations depressed both excitatory and inhibitory postsynaptic potentials in a concentration-dependent manner. On the other hand hyperpolarizing responses to exogenous gamma-aminobutyric acid (GABA) in the presence of bicuculline were suppressed by halothane, but depolarizing responses to L-glutamate were not altered. The results indicate that the depressant action of the anesthetic on the excitatory postsynaptic potential (EPSP) may occur presynaptically, whereas the blocking action on the inhibitory postsynaptic potential (IPSP) may occur postsynaptically.


Properties of depolarizing bipolar cell responses to central illumination in salamander retinal slices. The voltage and current responses of depolarizing bipolar cells to central illumination were studied by means of whole-cell recording in retinal slices of the larval tiger salamander, Ambystoma tigrinum. To stabilize the responses, it was necessary to limit exchanges between the cytosol and the solution in the patch pipette by reducing the diameter of the pipette tip opening. The current-voltage relationship of the cell membrane in darkness displayed a strong outward rectification, and the inward current evoked by light could be consistently reversed by depolarization only when tetraethylammonium was added to the pipette solution. As a result of the membrane non-linearity, increases in the intensity of bright lights caused relatively smaller amplitude increases in the voltage than in the current responses and the latter had a proportionally smaller after-effect. With larger pipette tip openings, the cytosol equilibrated with the pipette filling solution. Under these conditions the light-evoked responses gradually became slower and acquired an on-off pattern, their final amplitude and polarity being determined by the ratio of the chloride concentrations on each side of the cell membrane. This finding is interpreted as revealing the existence of two response components: a chloride-dependent on-off increase in conductance and a faster depolarizing input that was lost through diffusional exchange. Addition of GTP and ATP to the electrode filling solution had a stabilizing effect on the labile component, whether or not cyclic GMP was also included. Observations on the magnitude of the conductance changes and on diphasic reversals indicate that the labile response component, presumably representing direct input from photoreceptors, is caused by an increase in conductance. The resulting inward current was still present at a low intracellular chloride concentration and may be assumed, therefore, to be carried by a cation influx.


Localization and characterization of diazepam-binding inhibitor (DBI)-like peptides in the brain and pituitary of the trout (Salmo gairdneri). The distribution of diazepam-binding inhibitor (DBI)-like peptide(s) in the brain and pituitary of the trout was determined by the indirect immunofluorescence technique using an antiserum raised against synthetic rat octadecaneuropeptide (ODN). Numerous immunoreactive perikarya and processes were observed in the basal hypothalamus, within the pars lateralis of the nucleus lateralis tuberis. In the pituitary, ODN-immunoreactive processes were visualized in the neurohypophysial tract, projecting into the pars intermedia, and the pars distalis. Reverse-phase high-performance liquid chromatography combined to radioimmunoassay quantification was used to characterize the DBI-related material in tissue extracts. In both pituitary and hypothalamic extracts, the major immunoreactive form eluted with a retention time higher than that of rat ODN. In the hypothalamus, a minor peak co-eluting with the synthetic ODN standard was also resolved. The existence of peptides related to mammalian DBI in the hypothalamo-hypophysial complex of the trout suggests these neuropeptides may participate in the control of pituitary hormone release.


Acetylcarnitine, carnitine and glucose diminish the effect of muscarinic antagonist quinuclidinyl benzilate on striatal acetylcholine content. The content of acetylcholine (ACh) in the striatum, brain cortex and hippocampus of rats was lowered 20-180 min after intraperitoneal injection of the muscarinic antagonist quinuclidinyl benzilate (QNB). The depletion of ACh content in the striatum was diminished in animals treated with a single dose of acetyl-L-carnitine, L- or D,L-carnitine, or D-glucose. It is likely that QNB stimulates ACh release by blocking presynaptic muscarinic autoreceptors and that acetylcarnitine, carnitine and glucose support the resynthesis of ACh by increasing the availability of acetylcoenzyme A. They do not have the same consistent effect in the brain cortex and hippocampus; this difference may be related to the lower turnover rate of ACh and to the difference in the anatomical arrangement of cholinergic structures in these parts of the brain.


Effects of lesions of the optic nerve, optic tectum and nervus terminalis on rod precursor proliferation in the goldfish retina. Teleost retinas grow throughout life by proliferation of neuroblasts at the retinal margin and dedicated rod precursors in the outer nuclear layer. Mechanisms regulating this proliferation are largely unknown. Previous investigators observed that rod precursor replication, as detected by incorporation of radioactive thymidine into cells of the outer nuclear layer, is enhanced after optic nerve crush. We attempted to determine whether this was due to severing of the retinopetal (nervus terminalis, n.t.) or retinofugal (retinal ganglion cell) axons in the optic nerve of the goldfish, Carassius auratus. In the first series of experiments, we ablated unilaterally the optic nerve, olfactory bulb (containing n.t. ganglia), or optic tectum (containing retinal ganglion cell axons and n.t. collaterals). Rod precursor proliferation increased dramatically in both retinas as soon as 5 days after surgery; in addition, the numbers of dividing cells were greater in the ipsilateral retina 10-15 days after optic nerve crush or tectal ablation and in the contralateral retina 20-25 days after olfactory bulb ablation. These observations are not accounted for by the known projections of retinal ganglion cells, but are consistent with the projections of the n.t. In the second series of experiments, n.t. projections to the brain and retina were severed bilaterally 7-8 weeks before the unilateral optic nerve crush or hemitectal ablation. Rod precursor proliferation increased as before, but the quantities of dividing cells were always equal in both retinas. We conclude that the n.t. may modulate rod proliferation locally and that injury to (some) brain regions may cause release of mitogens that affect rod precursors in both retinas.


Does the release of vasopressin within the supraoptic nucleus of the rat brain depend upon changes in osmolality and Ca2+/K+? Microdialysis in conjunction with a highly sensitive radioimmunoassay was used to monitor the in vivo release of arginine vasopressin (AVP) within the supraoptic nucleus (SON) of the rat brain (n = 70). Thirty-min dialysates were collected in urethane-anesthetized animals before, during and after hypertonic or hypotonic pulses were delivered via the probe. As compared to artificial cerebrospinal fluid (aCSF)-perfused controls, 1M aCSF given over a period of 210 min resulted in an increased intranuclear AVP release which, however, reached its peak only in the post-stimulation period, i.e. after replacement of hypertonic with isotonic aCSF again (rebound phenomenon). Hypertonic (0.5 M, 1 M or 2 M) pulses given 150 min after the first (1 M) pulse resulted in 3 different rebound responses: a marked decrease (to 25.3%, P less than 0.001), no change or slight increase (132%, n.s.). As shown by the response to correction of the hypertonicity to normal as well as by perfusion of hypotonic aCSF (0.01 M), release of AVP within the SON appears more responsive to a reduction than to an elevation in the osmolality of aCSF. Omission of Ca2+ from and addition of EGTA to the aCSF decreased the osmotically stimulated, but not the basal AVP release. If K(+)-hypertonic aCSF was used, however, basal AVP levels increased significantly; in contrast, the rebound increase failed to differ from aCSF-perfused controls. It is concluded from these findings that release of AVP from intact neuronal structures in the SON is responsive to changes in osmolality with an apparent greater sensitivity to decrease in osmolality.


Medullary expiratory neurons in the decerebrate rat: an intracellular study. Intracellular recordings and labelings with horseradish peroxidase (HRP) of expiratory (E) neurons were performed in decerebrate, paralyzed, and ventilated rats. A total of 37 neurons were recorded, from which 4 cells and 1 axon were labeled. They were located in two regions of the ventrolateral medulla. One was in the rostral portion of the nucleus ambiguus just caudal to the facial nucleus, and the other in the nucleus retroambiguus at the level of the caudal medulla. These expiratory neurons had rhythmical changes in membrane potential similar to those reported in cat, i.e., a depolarization in the intervals between phrenic bursts which evolved in an augmenting (E-aug, n = 15), or bell-shaped or 'plateau' (E-all, n = 22) pattern until a rapid hyperpolarization at the start of inspiration. Both types were hyperpolarized during inspiration by chloride-dependent, inhibitory postsynaptic potentials (IPSPs) which were demonstrated in 17 neurons (10 E-aug and 7 E-all) from which reversal was obtained. Such IPSPs also existed during post-inspiration (stage I of expiration) in 4 of the 10 augmenting E neurons. They were identified by antidromic stimulation or HRP labeling, or both, as bulbospinal neurons (n = 2), cranial motoneurons (n = 4), or not antidromically activated (NAA) neurons (n = 31). All the identified bulbospinal neurons and the motoneurons exhibited an E-all pattern. The expiratory neurons of the caudal medulla had various projections as demonstrated with HRP labeling: one bulbospinal neuron with ipsilateral axon giving off intramedullary collaterals, and NAA neurons with rostral medullary projections or with axons crossing the midline.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of superior colliculus lesions on sensory unit responses in the intralaminar thalamus of the rat. The effects of kainic acid lesions of the intermediate and deep layers of the superior colliculus on the sensory input to the intralaminar thalamus of the rat were determined. Ipsiversive circling and contralateral sensory neglect were consistently seen after lesion placement. Two to 7 days later, the intralaminar thalamus was systematically explored for extracellular mechanoreceptive unit responses to high threshold and low threshold stimuli. On the side ipsilateral to the lesion the number of responsive units was reduced by 51%. The loss was particularly marked for nociceptive units (80%), and low threshold and complex units with orofacial receptive fields (73%). This effect may involve a partial deafferentation of the intralaminar thalamus as well as altered excitatory thresholds of thalamic neurons. It is suggested that the functionally distinct direct tectothalamic projection as well as the indirect tecto-reticulo-thalamic pathway are implicated.


Effects of PVN lesions on the responsiveness of female rats to estradiol. Previous research has shown that the paraventricular nucleus of the hypothalamus (PVN) is an important site of action for the effects of estradiol on feeding behavior. The recent finding that estrogenic stimulation of the PVN lowers food intake without inducing lordosis suggests that the effects of estradiol on feeding and sexual behaviors are organized separately within the brain. Whether the effects of estradiol on food intake can be attenuated by PVN lesions is therefore a question of practical and theoretical interest. In this experiment we examined the behavioral responsiveness of females with PVN lesions to peripheral treatment with estradiol. 32 adult, female rats received either bilateral or sham lesions of the PVN. All subjects were ovariectomized 2 weeks after the lesion. 2 Weeks following ovariectomy, half of the animals were injected with 2 micrograms of estradiol benzoate (EB) for 3 days, and half were injected with the oil vehicle. 10 days later, the treatment conditions for each subject (oil or EB) were reversed. Histological analysis indicated that 9 females had bilateral lesions of the PVN and 4 had bilateral lesions of the dorsomedial nucleus of the hypothalamus (DMN); 11 animals received sham lesions. Compared with oil treatment, EB injections significantly lowered water intake and body weight gain in all groups. However, food intake was suppressed in the DMN and sham but not in PVN-lesioned females. In addition, statistical analyses indicated that EB treatment induced similar levels of female sexual behavior in all groups. Thus, PVN lesions did not interfere with the ability of estradiol to stimulate lordosis.(ABSTRACT TRUNCATED AT 250 WORDS)