Increased levels of neuronal thread protein in cerebrospinal fluid of patients with Alzheimer's disease.

Neuronal thread protein is a recently characterized, approximately 20-kd protein that accumulates in brains with Alzheimer's disease (AD) lesions. This study examined whether concentrations of neuronal thread protein (NTP) were also increased in the cerebrospinal fluid (CSF) of individuals with probable (clinically diagnosed) and definite (histopathologically proved) AD. Using a highly sensitive three-site monoclonal antibody-based immunoradiometric assay, we measured NTP concentrations in CSF from 84 patients with probable AD and mild dementia (duration, 4.05 +/- 0.36 years), 45 with Parkinson's disease and minimal or no dementia (duration, 4.73 +/- 0.78 years), 73 with multiple sclerosis, and 73 nondemented control subjects. NTP concentrations were also measured in postmortem ventricular fluid and temporal lobe neocortex extracts from 31 subjects with histopathologically proved AD and 14 age-matched control subjects. The mean concentration of NTP in the CSF was higher in AD (4.15 +/- 0.25 ng/ml; 95% confidence interval [CI] limits, 3.65-4.65) than in Parkinson's disease (1.96 +/- 0.16 ng/ml; 95% CI, 1.65-2.27), multiple sclerosis (1.6 +/- 0.14 ng/ml; 95% CI limits, 1.33-1.88), or control subjects (1.27 +/- 0.06 ng/ml; 95% CI limits, 1.15-1.40) (p < 0.001). In addition, 70% of the patients with probable AD had concentrations of NTP in CSF that were higher than 2.5 ng/ml (> upper 99% CI limit in the control group), compared with 23% of Parkinson's disease patients, 11% of multiple sclerosis patients, and 4% of control subjects. The mean concentrations of NTP in the ventricular fluid and brain tissue from individuals with documented AD and end-stage dementia were threefold higher than the levels detected in the CSF from the remaining patients with probable AD and mild dementia. Moreover, of 9 patients with AD, postmortem brain and CSF manifested 5- to 50-fold higher levels of NTP compared with the CSF samples obtained an average of 6 years earlier. These findings demonstrate that NTP levels are elevated in the CSF of individuals with AD and that NTP levels in the CSF increase strikingly with progression of dementia and neuronal degeneration.


Decrease of D2 receptors indicated by 123I-iodobenzamide single-photon emission computed tomography relates to neurological deficit in treated Wilson's disease.

Single-photon emission computed tomography with 123I-iodobenzamide, a dopamine D2 receptor antagonist, was employed to study dopamine D2 receptor densities in 17 patients with biochemically proved Wilson's disease and stable neurological status with therapy and in 5 age-matched control subjects. Of the 17 patients with Wilson's disease, 5 were neurologically asymptomatic, 3 had cerebellar signs, 1 exhibited a mild parkinsonian syndrome, 7 showed a parkinsonian syndrome and cerebellar signs, and 1 had generalized dystonia and a parkinsonian syndrome. In 5 age-matched control subjects specific isotope binding as calculated by the basal ganglia to frontal cortex ratio was 1.57 +/- 0.04 (mean +/- standard deviation). The ratio in patients with Wilson's disease ranged from 1.56 +/- 0.05 (n = 5, asymptomatic patients) to 1.17 +/- 0.02 (n = 4, marked neurological impairment). We observed an almost linear correlation between the reduction of 123I-iodobenzamide (IBZM) binding and the severity of neurological signs at the time of IBZM-SPECT (correlation coefficient, -0.84; p < 0.01). We suggest that the reduction of postsynaptic striatal dopamine D2 receptors as detected by IBZM-SPECT reflects striatal neuronal damage in Wilson's disease.


Impaired activation of the supplementary motor area in Parkinson's disease is reversed when akinesia is treated with apomorphine.

Using positron emission tomography (PET) we previously showed that activation of the putamen, supplementary motor area, and cingulate cortex is impaired in patients with Parkinson's disease (PD) when they are off treatment and perform volitional motor tasks. Evidence suggests that these areas are involved in the generation of internally cued movements in normal subjects. We have now studied the effect of the dopamine agonist apomorphine on cerebral activation when used to treat the akinesia of PD. Regional cerebral blood flow was measured using C15O2 PET in PD patients at rest and when performing paced joystick movements with the right hand in one of four freely chosen directions. All patients used apomorphine regularly, and were studied before treatment, while still "off" but receiving a subcutaneous apomorphine infusion, and when switched "on" with apomorphine. Significant increases in regional cerebral blood flow were determined using statistical parametric mapping. Under resting conditions apomorphine had no effect on focal or global cerebral blood flow. Seven patients with PD performed the motor task adequately in the "off" and "on" states. This group of subjects demonstrated impaired activation of the supplementary motor area and contralateral putamen in the "off" state. Activation of the supplementary motor area significantly improved when the akinesia was reversed with apomorphine. We conclude that the concomitant improvement of supplementary motor area activation and motor function in apomorphine-treated patients with PD provides further evidence for the role of this structure in generating motor programs.


Using gadolinium-enhanced magnetic resonance imaging lesions to monitor disease activity in multiple sclerosis.

The highly variable clinical course and the lack of a direct measurement of disease activity have made evaluation of experimental therapies in multiple sclerosis (MS) difficult. Recent studies indicate that clinically silent lesions can be demonstrated by magnetic resonance imaging (MRI) in patients with mild relapsing-remitting MS. Thus, MRI may provide a means for monitoring therapeutic trials in the early phase of MS. We studied 12 patients longitudinally for 12 to 21 months with monthly gadolinium (Gd)-enhanced MRIs. The data have been used to identify the most effective design of a clinical trial using Gd-enhanced lesions as the outcome measure. Frequent ( > 1/mo) Gd-enhancing lesions were observed in 9 of the 12 patients, indicating that the disease is active even during the early phase of the illness. The frequency of the lesions was not constant; there was marked fluctuation in lesion number from month to month. However, the magnitude of the peak number of lesions and the frequency of the peaks varied among patients. Because of this variability, the most effective use of Gd-enhancing lesions as an outcome measure in a clinical trial was a crossover design with study arms of sufficient duration to allow accurate estimation of lesion frequency. Monitoring Gd-enhancing lesions may be an effective tool to assist in the assessment of experimental therapies in early MS.


Early childhood hepatocerebral degeneration misdiagnosed as valproate hepatotoxicity.

Four unrelated children were thought to have valproate-associated hepatotoxicity. They presented with recurrent partial secondarily generalized status epilepticus and epilepsia partialis continua followed by mental and motor regression. Despite treatment with multiple antiepileptic medications, they continued to have seizures. After initiation of valproic acid (VPA), all 4 manifested liver failure within 3 months. Two of these children each had 1 sibling who was not exposed to VPA, but who developed the same clinical picture including liver failure. At the time of autopsy, all 6 children had similar neuropathological findings with focal areas of spongiosis and neuronal loss, diffuse gliosis, and Alzheimer type II cells. One VPA-treated patient underwent a successful liver transplantation only to die from relentlessly progressive neurological deterioration. We propose that many of the reported patients with VPA-associated hepatotoxicity represent undiagnosed patients with early childhood hepatocerebral degeneration, the Huttenlocher variant of Alpers' syndrome. This disease manifests by obstinate partial seizures, recurrent partial secondarily generalized status epilepticus, epilepsia partialis continua, psychomotor deterioration, and hepatic dysfunction that is exacerbated by VPA administration. The accelerated demise from liver failure in the nontransplanted patients before the central nervous system pathology fully evolves makes the diagnosis of this rare condition difficult. The occurrence of disease in the unexposed siblings suggests recessive inheritance.


Continuous lisuride effects on central dopaminergic mechanisms in Parkinson's disease.

Effects of the long term, continuous administration of a dopamine agonist on motor response complications attending levodopa therapy were studied in 7 patients with advanced Parkinson's disease under controlled conditions. After a 3-month round-the-clock infusion of lisuride, the duration of antiparkinsonian action of levodopa increased by approximately 90%, and the therapeutic window for the acutely administered dopamine precursor widened by > 300%. These benefits were more than three times greater than those produced by 9 days of continuous levodopa administration. In contrast to the effects on levodopa pharmacodynamics, the continuous infusion of lisuride did not prolong its action, suggesting a lisuride effect on presynaptic as well as postsynaptic dopaminergic mechanisms. These results lend further support to the view that continuous dopamine replacement ameliorates motor fluctuations and peak-dose dyskinesias that complicate standard levodopa regimens. Our findings further suggest that alterations at both presynaptic and postsynaptic levels contributing to these motor complications tend to normalize with the more physiological stimulation afforded by continuous replacement strategies, especially when given chronically.


Platelet mitochondrial function in Parkinson's disease. The Royal Kings and Queens Parkinson Disease Research Group.

There is increasing evidence that defective function of the mitochondrial enzyme NADH CoQ reductase (complex I) is involved not only in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) toxicity, but also in idiopathic Parkinson's disease (PD). Complex I deficiency has been identified in PD substantia nigra and appears to be disease-specific and selective for the substantia nigra within the central nervous system. We describe a method for preparation of an enriched mitochondrial fraction from 60 mL blood. Using this technique, we analyzed respiratory chain function in 25 patients with PD and 15 matched control subjects. We confirm a previous report of a specific complex I deficiency in PD platelet mitochondria. Although there was a statistically significant decrease in complex I activity in the PD group compared with the control group (p = 0.005), the defect was mild (16%); it was not possible to distinguish PD from control values on an individual basis. This deficiency is not detectable in platelet whole-cell homogenates, presumably reflecting the relative insensitivity of this preparation and the limited decrease in complex I activity in PD. The presence of a mild complex I defect in platelets together with a more severe defect in substantia nigra suggests either that the pharmacological characteristics shared by these two tissues render them susceptible to a particular toxin or toxins, or that the defect is widely distributed and other biochemical events enhance the deficiency in substantia nigra.(ABSTRACT TRUNCATED AT 250 WORDS)


Temporal lobectomy for uncontrolled seizures: the role of positron emission tomography.

We evaluated the role of positron emission tomography (PET) with [18F]deoxyglucose (FDG) (FDG-PET) for planning surgery in 53 patients who had temporal lobectomy for uncontrolled seizures at National Institutes of Health from 1981 to 1990. Investigators blinded to PET data used results of telemetered video-electroencephalographic ictal monitoring and other standard criteria to decide whether subdural electrodes (22 patients, i.e., the "invasive" group) should be implanted or surgery performed. PET scans were analyzed using a standard regional template. Mean lateral but not mesial temporal asymmetry was significantly higher in patients who became seizure free (p < 0.03). Patients with > or = 15% hypometabolism were significantly more likely to be seizure free in the entire study population and the invasive subgroup. Visual identification of hypometabolism was less accurate. When a clear temporal ictal surface electroencephalographic focus was present, FDG-PET provided less additional information. FDG-PET may be particularly valuable if the surface electroencephalographic scan is nonlocalizing. In addition to helping to identify the seizure focus, it may allow limitation of invasive electrode placement to those necessary for functional mapping. When PET is used to identify epileptic foci, quantitative measurements of asymmetry should be made.


The antiparkinson efficacy of deprenyl derives from transient improvement that is likely to be symptomatic.

We undertook an analysis of the hazard functions derived from results published by the Parkinson Study Group following their investigation of deprenyl. Our findings indicate that the action of deprenyl is transient rather than sustained. We also infer that this effect may be mediated through alleviation of symptoms rather than by neuroprotection.


The free radical hypothesis in idiopathic parkinsonism: evidence against it.

The free radical hypothesis for the pathogenesis of idiopathic parkinsonism (Parkinson's disease) has many similarities to the argument invoking an autoimmune mechanism. In both cases, cellular and molecular machinery that might be involved in neuronal destruction have been demonstrated. In recent years, the free radical hypothesis has become particularly fashionable; several workers have reported observations which, they infer, support the notion that damage by free radicals is a major factor in the underlying disease process. There is, however, no conclusive evidence that free radicals play a prominent role in the causal chain of events that leads to idiopathic parkinsonism; several findings may be construed as evidence against such a contention.


The oxidant stress hypothesis in Parkinson's disease: evidence supporting it.

Oxidant stress, due to the formation of hydrogen peroxide and oxygen-derived free radicals, can cause cell damage due to chain reactions of membrane lipid peroxidation. Because the substantia nigra is rich in dopamine, which can undergo both enzymatic oxidation via monoamine oxidase and nonenzymatic autoxidation, hydrogen peroxide and oxyradicals (superoxide anion radical and hydroxyl radical) are generated in this midbrain nucleus. Although proof that oxidant stress actually causes the loss of monoaminergic neurons in patients with Parkinson's disease is lacking, there is a considerable body of evidence from studies in both animals and humans that support the concept. (1) Neurotoxins that selectively destroy the dopaminergic neurons in the nigra, such as 6-hydroxydopamine and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), appear to act via oxidant stress. (2) The substantia nigra of patients with Parkinson's disease reveals evidence of oxidant stress by the findings of increased lipid peroxidation and decreased reduced glutathione. (3) Total iron is increased and ferritin is reduced in the substantia nigra pars compacta in patients with Parkinson's disease. This combination suggests that this transition metal is in a low molecular weight form, capable of catalyzing nonenzymatic oxidative reactions, especially the conversion of hydrogen peroxide to hydroxyl radical, which is the most reactive of the oxygen radicals. (4) Neuromelanin, a product of dopamine autoxidation, can serve as a reservoir for iron, promoting the generation of oxyradicals. (5) Antioxidant defense mechanisms appear to be reduced in the parkinsonian substantia nigra with the findings of decreased activities of glutathione peroxidase and catalase.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression of 65-kd heat shock proteins in the inflammatory myopathies.

In normal muscle, 65-kd heat shock proteins (hsp) were detected on capillary endothelial cells, the mural elements of larger vessels, and some intracellular organelles, probably mitochondria. In the inflammatory myopathies, the 65-kd hsp were detected on inflammatory cells, degenerating and regenerating fibers, and on many but not all nonnecrotic muscle fibers invaded by T cells. The expression of the 65-kd hsp may be an immune-nonspecific response to cellular "stress," but hsp determinants could possibly also serve as autoantigen(s) recognized by autoreactive T cells.


Familial rectal pain: a type of reflex epilepsy?

We studied 2 members of a family suffering from paroxysmal attacks elicited by tactile stimuli. The attacks consist of burning pain of the stimulated body part, followed by either complete collapse or tonic posturing. Noxious stimuli provoke episodes regardless of their somatic location, whereas it is only necessary for nonnoxious stimuli to be applied to specific trigger zones, such as the rectum, to provoke attacks. Episodes are most commonly precipitated by bowel movement, leading to extreme fear of defecation and resultant fecal retention. An ictal electroencephalographic video recording revealed only slowing of the background; however, serum prolactin was significantly elevated postictally. The attacks were completely suppressed by carbamazepine and resumed on discontinuing the medication. These attacks may represent a form of reflex epilepsy manifested by autonomic nervous system dysfunction.


Chronic encephalitis (Rasmussen's syndrome) and ipsilateral uveitis.

Two children are reported with acute uveitis during the early progressive phase of chronic encephalitis (Rasmussen's syndrome). In both children, the side of the uveitis was ipsilateral to the side of cerebral inflammation, although in 1 child there were milder inflammatory changes in the contralateral eye. This association adds weight to the viral hypothesis of chronic encephalitis and raises the possibility of primary ocular infection and neurotropic spread to the brain.


Psychological effects of sodium valproate and carbamazepine in epilepsy.

Information from standardised tests of intelligence, school attainments, attention, memory and visuomotor function, together with parent and teacher questionnaire information about various aspects of behaviour, was obtained for 63 schoolchildren with newly diagnosed epilepsy before treatment with sodium valproate or carbamazepine, and again at intervals for a total period of 12 months. The same information was collected on 47 matched controls. The children with epilepsy represented those under non-specialised paediatric care. The result showed that both drugs were effective in most cases at modest dosage without causing notable psychological effects 12 months into treatment. Modest and temporary adverse cognitive effects seen earlier in treatment could have been the result of uncontrolled seizure discharge. Improved function was the same in children with epilepsy and controls. Some psychological abnormalities in the children with epilepsy were evident before treatment suggesting early unwanted effects of the epileptic process itself.


Foregut motor function in chronic renal failure.

In children with chronic renal failure (CRF) anorexia, nausea, and vomiting are common yet poorly understood symptoms. We studied oesophageal and gastric motor function in 12 children (age 7 months-6.8 years) with severe CRF not undergoing dialysis who had persistent anorexia and vomiting. Eight of 12 patients had significant gastro-oesophageal reflux (reflux index 5.2% to 21.9%, mean 11.3%; controls < 5%), 7/10 had altered gastric half emptying times (T1/2) for 5% glucose or milk (glucose meal--controls: 8-14 min, two CRF patients: 18-25 min; milk meal--controls: 48-72 min, five CRF patients 27, 28, 82, 83, and 110 min). Gastric antral electrical control activity was abnormal in 6/11 patients, with different types of gastric dysrhythmias whereas the remainder and controls showed a regular dominant frequency of 0.05 Hz. In 7/9 patients fasting serum gastrin concentration was raised (53 to > 400, mean 168 pmol/l, controls < 40 pmol/l). All CRF patients with anorexia and vomiting had one or more disorder of foregut motility. The nature and variety of the motor disorders and the raised concentrations of circulating gastrin suggest that the normal environment generated by CRF affects the function of the smooth muscle of the foregut.


Ultrasonographic assessment of the extent of hepatic steatosis in severe malnutrition.

Ultrasonographic, blinded assessment was made of the extent of hepatic steatosis in 55 children with severe malnutrition: undernutrition (n = 6), marasmus (n = 18), marasmickwashiorkor (n = 17), and kwashiorkor (n = 14). The children were examined on admission, in early recovery (considered as baseline), and again at discharge. Eleven healthy control children and eight of the previously malnourished children were studied as comparison groups. Both oedematous and non-oedematous malnourished children had significantly more steatosis than the comparison groups at each time. Children with oedematous malnutrition had significantly greater steatosis than non-oedematous children at admission. Half of the non-oedematous malnourished children had appreciable hepatic steatosis at both admission and at baseline. Hepatic fat was only slowly mobilised. The rate constant was 1.4 +/- 0.3%/day. One quarter of the children did not change steatosis grades during the period they were in hospital. There was no overall correlation between the extent of steatosis and liver size. Hepatic steatosis in childhood malnutrition is not confined to oedematous children: it is frequently present in marasmic and undernourished children. Its extent is not necessarily related to the degree of hepatomegaly and accumulated lipid is only slowly mobilised.


Adult height in constitutionally tall stature: accuracy of five different height prediction methods.

The accuracy of height predictions at various ages based on five different methods (Tanner-Whitehouse mark I; Tanner-Whitehouse mark II; index of potential height; Bayley-Pinneau; Roche-Wainer-Thissen) was compared at yearly intervals with final height achieved in 32 boys (78 predictions) and 100 girls (227 predictions) with constitutionally tall stature. The boys were initially seen at a mean (SD) chronological age of 12.5 (3) years whereas the mean chronological age in girls was 11.8 (2.1) years. In tall boys Tanner-Whitehouse mark II gives a good estimation of final height up to the bone age of 13 years with a mean overestimation of 1 cm. The overestimation of final height is higher in the bone age groups 13-14 years (2.7 cm) and 14-15 years (3.4 cm) mainly due to the tall boys with a height greater than 3 SD scores. Up to the bone age of 12 years the final height is massively overestimated by the Bayley-Pinneau method but this method give relatively accurate estimations thereafter. The estimated confidence limits are large (+/- 8 cm) for the two methods up to a bone age of 15 years. In tall girls the Tanner-Whitehouse mark II method was accurate from bone age nine to 12 years but overestimated final height in the bone age groups 12-13 years and 13-15 years by a mean of 1.8 and 1.4 cm respectively. The Bayley-Pinneau method overestimated final height in the bone age groups 12-14 years whereas the height predictions are accurate thereafter. Up to a bone age of 13 years the estimated confidence limits for the two methods are large, +/- cm, but tend to improve thereafter. It is concluded that there is no best or most accurate method for predicting adult height in tall children. There are methods of first choice differing with respect to sex and bone age. In addition, correcting factors may improve their accuracy and correct their tendency to overestimate or underestimate adult height.


Incidence of hepatic hamartomas in tuberous sclerosis.

Hepatic hamartomas were thought to be a rare finding in patients with tuberous sclerosis. The purpose of this study was to assess their incidence in children with tuberous sclerosis and to review the literature. During 1984-90 we examined 51 children by ultrasonography; there were 25 boys and 26 girls. Their age ranged from 3 months to 18 years. Liver hamartomas were seen in 12 (23.5%) of the children, more often in girls than boys (5:1). Their incidence increased with age reaching 45% in children over the age of 10. They did not produce any symptoms of hepatic dysfunction. Our study and review of reported cases prove that hepatic hamartomas are a common finding in patients with tuberous sclerosis and may be very helpful in providing a more accurate diagnosis and consequently help in genetic counselling.


Increased resting energy expenditure in childhood asthma: does this contribute towards growth failure?

In order to determine whether or not there was a relationship between disorders of growth in children suffering from asthma and either increased resting energy expenditure or inadequate energy intake, a group of 34 children suffering from perennial symptoms were studied. A control group matched with the asthmatic children for sex and fat free mass were similarly studied. The children kept seven day records of weighed food intake. Basal metabolic rate was measured on one occasion in the fasted state by means of indirect calorimetry using the ventilated hood technique. The asthmatic children kept a 28 day record of peak expiratory flow rates, asthma symptoms, and medication usage. The asthmatic children expended significantly more energy at rest than their matched controls in absolute terms (14%). There was no correlation between height or height SD score and any parameter of energy balance. The causes of these finding are as yet speculative.


Defective alloantigen-presenting capacity of 'Langerhans cell histiocytosis cells'.

The functional activity of skin cells derived from an infant who died of multisystem Langerhans cell histiocytosis (LCH) was examined. Involved and non-involved skin was obtained at postmortem examination within three hours of death; normal epidermal Langerhans cells and 'LCH cells' were separated by means of dispase digestion. The functional activity of different populations of CD1a positive cells was assessed using the conventional six day allogeneic mixed cell reaction. Compared with Langerhans cells from a healthy control, LCH cells showed minimal functional activity. However, Langerhans cells from non-involved skin showed normal and Langerhans cells overlying involved skin showed augmented functional activity. These findings suggest that LCH is a disease in which abnormal Langerhans cells accumulate and/or proliferate in various tissues but it does not affect the entire Langerhans cell population.


Interleukin-2 in relation to T cell subpopulations in rheumatic heart disease.

Interleukin-2 (IL-2) and T cell subpopulations were evaluated in children with rheumatic heart disease (RHD). Three groups were included: 13 patients with active RHD, 12 with non-active RHD, and 14 control children. Serum IL-2 and T cell subpopulations were measured by radioimmunoassay and monoclonal antibodies respectively. Patients with active RHD showed a significant increase in IL-2 concentrations and helper:suppressor (H:S) ratio compared with controls with a mean (SEM) IL-2 of 3.48 (0.62) v 1.26 (0.16) U/ml and H:S ratio 2.31 (0.14) v 1.66 (0.04). There was a significant decrease in T suppressor (CD8+) and pan T (CD3+) cells compared with controls with a mean (SEM) for CD8+ of 23.75 (1.19) v 32.23 (0.56)% and CD3+ of 79.55 (0.94) v 85.00 (0.11)%. Patients with non-active RHD showed a significant decrease only in the CD3+ cells (78.20 (0.20)%) when compared with controls. A deficiency of CD3+ cells is a constant finding in patients with RHD, whether the disease is active or not. There was a significant increase in IL-2 concentration with a significant decrease in CD8+ cells in patients with active RHD in comparison with the non-active group (mean (SEM) IL-2 of 3.48 (0.62) v 1.85 (0.24) U/ml and CD8+ of 23.75 (1.19) v 28.83 (1.91)%). Thus an increase in IL-2 and a decrease in CD8+ cells may be related to rheumatic activity. T helper (CD4+) cells did not differ significantly between groups.


Bronchoscopy in lipoid pneumonia.

Forcible administration of rendered animal fat to infants is a tradition in south western Saudi Arabia. Accidental inhalation may result in a resistant form of lipoid pneumonia. A series of 24 cases of lipoid pneumonia, 22 of which were diagnosed by bronchoscopy with bronchial lavage and microscopic examination of the aspirate, are reported. The technique is described briefly and the results analysed. A high index of suspicion together with bronchoscopy and bronchial lavage of all cases of resistant or recurrent pneumonia is essential in areas such as ours for diagnosis of this condition.


Causes of death in children diagnosed with non-Hodgkin's lymphoma between 1974 and 1985.

An investigation has been undertaken of 479 deaths occurring up to the end of 1990 among 883 patients diagnosed with non-Hodgkin's lymphoma from 1974 to 1985 who were included in the population based National Registry of Childhood Tumours. The objectives were to perform a descriptive analysis looking particularly at the deaths not directly due to non-Hodgkin's lymphoma, to determine the frequency of the different causes of death and to study the trends over time. Among the 476 patients with sufficient information for the cause of death to be established, these were: non-Hodgkin's lymphoma, 377 (79%); treatment related (other than second primary tumour), 86 (18%); second primary tumour, 10 (2%); and other, three (1%). The proportion of all deaths not directly due to non-Hodgkin's lymphoma increased from 15% for those diagnosed during 1974-6 to 32% for those diagnosed during 1983-5. Among the 86 treatment related deaths, the more precise causes were bacterial infections, 26 (30%); viral and other infection, 14 (16%); metabolic, 19 (22%); renal, eight (9%); anaesthetic related, seven (8%); respiratory, four (5%); cardiac, three (3%); graft versus host disease, three (3%); and other, two (2%). Treatment related deaths from infection accounted for 27 (6%) of all patients diagnosed in 1974-9, and 13 (3%) in 1980-5. Treatment related deaths not due to infection occurred in 23 (5%) of those diagnosed in 1974-9 and 23 (6%) in 1980-5. Five treatment related deaths, including four anaesthetic related deaths, were identified as avoidable. Some of the deaths from metabolic and renal disease may also have been avoidable. Only 11 deaths have been recorded more than five years after diagnosis, six being due to second primary tumours. As follow up is relatively short for patients diagnosed more recently, further deaths from second malignancies and treatment related cardiovascular problems may well occur. A substantial number of children with non-Hodgkin's lymphoma die to treatment related causes. Deaths from infection have decreased in line with the overall improvement in survival rates. Other treatment related mortality has remained constant. Further improvements in survival for childhood non-Hodgkin's lymphoma will depend on maintaining the fine balance between the therapeutic value of intensive treatment and its potential harmful effects.


Antiphospholipid antibody associated thrombosis in juvenile chronic arthritis.

A child with systemic onset juvenile chronic arthritis (JCA) who developed a bilateral femuropopliteal vein thrombosis after plaster immobilisation following a tibial fracture is described. When the thrombosis was diagnosed, antiphospholipid antibodies detected either as lupus anticoagulant and anticardiolipin antibodies were found. This suggests that short term prophylatic antithrombotic treatment should be considered in antiphospholipid antibody positive JCA patients who require immobilisation after fractures of demineralised bones.


Severe anaemia and ileocolic anastomotic ulceration.

Two children are described with anaemia from ileocolic anastomosic ulceration as a late complication of surgery in the newborn period. The anastomosis was revised in each case but in one child there was early recurrence of ulceration.


Intercurrent illness in inborn errors of intermediary metabolism.

Metabolic decompensation may occur in patients with disorders of intermediary metabolism during intercurrent illness. To prevent complications it is normal practice to change the diet to an 'emergency regimen'. The mainstay of this is a high carbohydrate intake, using soluble glucose polymer, given as frequent drinks by day and during the night. Additional therapy is given for some disorders. Practical details of the treatment are outlined.


Treatment of delinquents.

The medical role in the treatment of delinquency is a limited one. There is conflicting evidence as to whether treatment aims should be directed towards the individual, to the family, the institution, or the therapist. Nevertheless there seems to be a consensus of opinion that short term, focused therapies aimed at improving educational, vocational and social skills, possibly from a preschool age, are the most effective. Any treatment gains achieved while in residential care appear to be short lived. It therefore seems that this should be reserved for those individuals who commit repeated, violent crimes and for those from very damaging family backgrounds who repeatedly abscond or absent themselves from community based programmes.


Maternal immunization by husband's leukocytes for repeated fetal death associated with mild pre-eclampsia--case report with successful outcome.

We report a case of repeated fetal death at 31 gestational weeks associated with mild non-proteinuric pre-eclampsia and intrauterine growth retardation. After double intradermal immunisation with paternal leukocytes, a third pregnancy proceeded uneventfully until it ended at 38 weeks. Maternal anti-paternal blocking antibody activity was assessed by the erythrocyte antibody inhibition (EAI) test. Serologic testing revealed that the couple did not share HLA class I antigens. The mechanisms underlying the likely benefit from immunotherapy are discussed.


Listeria monocytogenes. The role of transabdominal amniocentesis in febrile patients with preterm labor.

Two women with preterm labor and intraamniotic infection with Listeria Monocytogenes are presented. In both patients, the prenatal diagnosis of Listeriosis was made by transabdominal amniocentesis. The immediate prominent observation was meconium staining of the amniotic fluid. We propose that an amniocentesis should be performed in women with premature labor and fever. If the amniotic fluid is meconium stained and the Gram stain examination reveals Gram positive rods, Listeria Monocytogenes should be suspected and the patient should be treated accordingly until the culture results are obtained.


An endoscopically assisted technique for construction of a neovagina.

The specific disadvantage of the commonly used Vecchietti method for reconstruction of the vagina in cases of vaginal agenesis is the need for laparotomy. We now describe a successful endoscopically assisted construction of a neovagina by a variation of the Vecchietti method. The method causes relatively little stress to the patient.


Density and distribution of Langerhans cells in the human uterine cervix.

Using a combination of immunohistochemistry and planimetric observation we report the density and distribution of Langerhans cells (LCs) in epithelium from the normal ectocervix and from the normal transformation zone (TZ). The density of LCs increased towards the basement membrane. Dendritic LCs aspect was more prominent in the superficial layers.


Structural changes of a devascularizated colonic graft when its replaces the rat uterine horn.

A one centimeter length of the left uterine horn in two groups of Wistar rats (n = 10) was replaced with normal (VAS group) or with denervated and devascularizated (NVAS group) colonic grafts. All animals maintained pregnancy in the right control horns but not in the grafted horns. At 40, 60 and 90 days after surgery, the light microscopic appearance of the autografts was studied. In the VAS group, and with respect to the last period, the number (5.5 +/- 0.7) and height (1.0 +/- 0.1 mm, P < 0.05 ANOVA) of the folds, the intestinal glands height (160.2 +/- 21.2 mu, P < 0.05 ANOVA) and the number of globet cells per gland (26.6 +/- 4.2, P < 0.05 ANOVA) had decreased in relation to the colon control (6.0 +/- 0.7, 1.4 +/- 0.1, 251.7 +/- 31.8, 42.6 +/- 5.2 respectively). A similar intestinal structure to that described above was observed in the anastomosis areas of the NVAS group, and therefore a decreased mucus production was maintained in this areas. No folds or intestinal glands were observed, but a monoestratified cubic epithelial cells type was observed along 63.7 +/- 4.1% of the NVAS colonic graft center. Under this epithelium a connective tissue, like a non-glandular submucosa, was obtained.


A controlled study of maternal serum concentrations of lipoproteins in pregnancy-induced hypertension.

Seventeen women with pregnancy-induced hypertension (PIH) and 17 controls with uncomplicated pregnancy who were matched for maternal and gestational age were investigated for lipid concentrations in whole serum and lipoprotein fractions. The triglyceride level in the low-density lipoprotein fraction was higher in PIH patients than in controls (P < 0.03). The differences between values of other parameters did not attain the level of statistical significance, although the tendency of the results was consistent with findings reported in other studies. The differences observed were small and the data exhibited wide variation. Therefore, it appears unlikely that serum lipid and lipoprotein concentrations could gain major clinical importance as diagnostic parameters in PIH.


Survey of computerized obstetric information systems in Austria.

The aim of this study was to evaluate obstetric electronic data processing (EDP) in Austria and to analyse its problems, advantages and acceptance in a single big obstetric department. We sent questionnaires to every obstetric department in the country. The overall response rate was 77% (73 departments). Only 24 (33%) were using computer aided documentation, but these covered 63% of deliveries in Austria. The proportionate times spent on documentation were 57% for physicians and 43% for midwives, with physicians playing a bigger role in larger departments using electronic documentation. Sixty-five percent of physicians and 31% of midwives readily accepted computerization. We also studied an obstetric department with over 3000 births per year. Twenty-five percent of the medical staff did not believe that computerization saved time, although they appreciated its value to administration and for producing printouts. Advantages in completeness (92%) and accuracy (76%) were recognized. After 6 month's use acceptance of EDP documentation improved significantly.


The relation between the results of cytophotometric examination of endometrial carcinoma and clinical course of these disease.

We made a prospective cytophotometric study of 48 cases of endometrial carcinoma. Thirty-two (66.7%) of the cases had diploid DNA content and the proportion of non-diploid tumors increased with the staging of tumors, lack of differentiation and depth of myometrial invasion. DNA content (AE), DNA malignancy grade (DNA-MG) and 5c exceeding rate were statistically significant in relation to the clinical course of the disease and the histological grade of differentiation. DNA parameters appear to be especially suitable for the objective assessment of malignancy.


Treatment of lymphangioleiomyomatosis by ovariectomy, interferon alpha 2b and tamoxifen--a case report.

The gender-specific prevalence of lymphangioleiomyomatosis (LAM) in premenopausal women suggests a hormonal etiology. Despite the antiestrogenic treatment (ovariectomy, tamoxifen) this disease is often refractory to therapy and almost inevitably leads to the patient's death. We describe a case where the antiproliferative effect of systemically applied interferon alpha 2b was successfully employed in addition to ovariectomy and the patient reached complete remission.


Expression of epidermal growth factor receptor after partial pancreatectomy in adult rats: an immunohistochemical study.

The expression and localization of epidermal growth factor (EGF) were investigated immunohistochemically using an anti-EGF receptor antibody in the pancreas of partial pancreatectomized and sham-operated adult rats. In the sham-operated pancreas, immunoreactive products against EGF receptor were only slightly positive in the pancreatic acinar cells. In the partial pancreatectomized pancreas on the fifth day after operation, EGF receptor immunoreactivity was intensely positive in the acinar cells, in some cells lining the intercalated ducts and some basal cells of the acinus, but it was not detected in the pancreas when exogenous EGF was given to the rat after partial pancreatectomy for three days. The present results suggest that EGF receptor is expressed in the regenerating pancreatic tissue, and EGF could be involved in the mechanism of pancreatic regeneration in rats.


[Study on 27 surgical cases of hypertrophic pyloric stenosis at Kyoto City Hospital].

Twenty-seven patients of hypertrophic pyloric stenosis operated on at our hospital between 1977 and 1991 were reviewed. The patients consisted of 22 boys and 5 girls, with males accounting for 81%. Seventeen of the 27 patients were the first children of their mothers. Vomiting was first noted between 10 and 58 days after birth with a mean of 26.5 days, and neonatal onset was observed in 67% of all patients. The body weight decreased after the onset in 17 patients, including 1 in which it decreased below the birth weight. Hypochloremia was the most frequent preoperative electrolyte imbalance, being observed in 41% of all patients. Alkalosis was noted in 17 of the 22 patients in which arterial blood gas analysis could be performed. The olive was palpated preoperatively in 24 (89%) of the 27 patients. The body weight increased in all patients after operation, and the mean daily increase was 26.7 g. The mean period of hospitalization after operation was relatively short at 8.3 days. Postoperative vomiting was observed in 16 patients (59%), with its mean duration being 26 days. All patients showed normal growth after operation, and no postoperative complications were noted.


Variations in warfarin concentration in blood and coagulant factors after artificial valve replacement.

A proper dosage of warfarin after artificial cardiac valve replacements has been determined as an indication of coagulation activity. In some cases, the coagulation activity cannot be maintained within the therapeutic range because Prothrombin time (PT) values deviated from Thrombotest (TT) values. Here we studied each relationship between warfarin concentrations in blood, coagulation activity, vitamin Ks and other coagulation-related agents which were measured in patients administered warfarin after artificial cardiac valve replacements and whose coagulation activity was maintained within a therapeutic range, and in patients whose PT values deviated from TT values. The obtained results were interpreted as follows. 1) There was a high correlation between values from Thromborel S (a clot method) and values from Chromoquick (a method using a synthesized substrate). Since TT values had little variations compared to PT values, the former was an excellent indication of coagulation activity. 2) There was no correlation between warfarin concentrations in blood and coagulation activity in the group in which warfarin concentrations in blood vary within a day (Group B); but there was highly reverse correlation in the group in which warfarin concentrations in blood vary with days (Group A). 3) There was good correlation between warfarin concentrations and PIVKA-II concentrations in blood in Group A. Further, the concentration of Gla-Protein C was one microgram/ml or less on the fourth day from the initiation of warfarin administration. 4) For example, a patient whose PT value of 67% deviated from TT value of 19% on the eleventh day from the initiation of warfarin administration had vitamin K1-epoxide concentration of 2.68 ng/ml and MK-7 concentration of 1.26 ng/ml on the same day, respectively, which were higher levels than those in Group A.


Effects of warfarin at beginning doses on patients with valve replacement.

Warfarin administered to patients with valve replacement at a dose of 0.1 mg/kg for 2 days after surgery, and blood levels of warfarin, TT, PT, HPT, total vitamin K and vitamin K1-epoxide were determined at various time intervals after the administration of the drug. The measurements were obtained from 15 patients: 8 patients with AVR, 6 patients with MVR, and 1 patient with DVR. The blood levels of warfarin reached a peak of 462 +/- 160 ng/ml 6 hours after the administration, TT, PT and HPT were 40% of the initial values 24 hours after the administration, and vitamin K1-epoxide was 1.28 +/- 0.30 ng/ml 24 hours after the administration. It is clear from the results that warfarin exhibits sufficient anticoagulant effects at a beginning dose of 0.1 mg/kg after surgery to prevent a sudden decrease in blood coagulation.


The efficacy of gemfibrozil therapy for raising high density lipoprotein levels.

Thirty subjects, 5 normotriglyceridemic (NTG) with low HDL cholesterol (HDL-C < 35 mg/dl) and 25 hypertriglyceridemic (HTG) with low and high HDL-C (HDL-C > 35 mg/dl) were selected fo this study. They were treated with gemfibrozil (600 mg BID) for 12 weeks. In both groups, gemfibrozil significantly reduced serum TG levels (p < 0.005), yet HDL-C increased significantly only in HTG patients (p < 0.005). The changes in HDL-C levels were highly variable (-40 to 50%) and appeared to be dependent on the levels of serum TG achieved during treatment. Based on post-treatment serum TG, the HTG patients were divided into 2 groups. Group 1 with serum TG of < 100 mg/dL and Group 2 with serum TG levels > 100 mg/dl. Significant post treatment increases in HDL-C were seen only in Group 1 (p < 0.005). The two groups had similar pretreatment serum TG and HDL-C levels but the LDL-C was significantly higher in Group 1 (p < 0.025). Pretreatment serum LDL-C also correlated positively with the increases in HDL-C during treatment (r = 0.51, p < 0.01, n = 25). Consequently, the patients were divided into three groups based on their initial serum LDL-C levels (Group 1: LDL-C < 130 mg/dl. Group 2: LDL-C, 130-159 mg/dl and Group 3: LDL-C > 160 mg/dl). The HDL-C levels increased significantly upon treatment only in Group 3. Pretreatment levels of serum TG and HDL-C were not significantly different among the three groups. Initial body weight (r = -0.43 p < 0.025, n = 30) and percent change in body weight during treatment (r = -0.47, p < 0.025, n = 30) correlated negatively with the percent reduction in serum TG. The change in body weight also showed significant negative correlation with the changes in HDL cholesterol (r = -0.48, p < 0.25, n = 30). We conclude that gemfibrozil is most effective in reducing serum triglycerides, LDL-C and increasing serum HDL-cholesterol in HTG patients who also have comparatively high initial LDL cholesterol levels (Fredrickson's type IIb phenotype). For effective improvement of HDL-cholesterol in most HTG patients, serum TG levels need to be lowered below 100 mg/dl. Furthermore, the benefit of gemfibrozil therapy may be significantly enhanced by weight loss during treatment.


[Morphologic changes of the brain in intracranial metastatic tumors].

18 brains of patients who died of intracranial metastases at the age of 27 to 72 years were morphologically studied. Pronounced oedema and the lack of noticeable macro- and microglia proliferation were found out at the border of metastases. Groups of cancer cells were observed at the distance from the main metastatic focus; these groups were considered as micrometastases.


[Morphology of bronchiolo-alveolar cancer].

The criteria of the morphological diagnosis of bronchiolo-alveolar carcinoma are presented on the material of 140 cases. Three main histological types are distinguished: clear-cell, dark-cell, polymorphic-cell. Anatomical forms of tumour growth are also identified: nodular, pneumonia-like, disseminated.


[Mucous membrane of the jejunum in allergy to cereal protein].

Biopsies of the jejunum from 61 children, 6 months-4 years of age, including 35 with allergy to corn protein, 18 with celiac disease and 8 children without stomach and intestine pathology were studied. Biopsies were fixed in 10% formalin or in Carnois fixative, the sections were stained with hematoxylin and eosin, 0.5% toluidine blue (pH 0.5), and immunoperoxidase method was used for revealing IgA. The structure of the mucosa was not changed in the allergic patients, the content of the granulated and ungranulated labrocytes was increased, their number being decreased in the elimination diet. The number of plasma cells was diminished, and in only few of them IgA was found in children with severe alimentary malabsorption. A selective deficiency of IgA was diagnosed which determines the therapy of such children. Hyperregenerative atrophy of the mucous membrane and predominance of degranulated cells over undergranulated were noted in celiac disease.


[Morphology of tuberculous meningitis in children].

25 observations of tuberculosis meningitis in children who died at the age from 3 months to 6 years with a clinical course from 5 days to 6.5 months were analyzed. Meningitis was not diagnosed clinically in 6 children. The inflammation was of an exudative or exudative-necrotic type at all periods of the disease. Specific features were: long-lasting macrophagal reaction, the presence of numerous plasma-cells (particularly at late stages of the disease), predominance of diffuse rather than granulomatous inflammation. Combination of delayed and immediate hypersensitivity reactions and possible organ specificity of the process may be involved in the pathogenesis of the disease.


[Morphologic features of the liver in sarcoidosis].

Granulomatous hepatitis with non-caseous epithelioid-cell granulomas (43 cases, 86%) and non-specific reactive hepatitis (7 cases, 14%) were observed in liver biopsies of 50 patients with sarcoidosis. Three types of granulomas are distinguished: macrophagal, florid and fading epithelioid-cell granulomas which are consequent stages of the granuloma transformation. Fibrosis arises in the macrophagal granuloma, is increasing in both epithelioid-cell granulomas and is ended by a formation of a small scar. Nonspecific liver changes were also found (hydropic and fat hepatocyte degeneration, hypertrophy and proliferation of the stellate reticulo-endotheliocytes, sinuses capillarization, lymphoid-histiocytic infiltration of the intralobular stroma and portal tract, step-like necrosis which are probably induced by the action of the sarcoidosis unknown etiological factor.


[Characteristics of primary glomerulonephritis (on the basis of kidney biopsies of the Pathology Department, I. M. Sechenov Moscow Medical Academy, from 1980 to 1989)].

On the basis of 1263 observations a relative incidence and clinical manifestations of main morphological forms of primary glomerulonephritis (PGN) are studied. Alterations in the clinical and morphological structure of primary glomerulonephritis are noted with nephrotic forms becoming more frequent and mesangiocapillary glomerulonephritis among patients with nephrotic and nephrotic-hypertension syndrome becoming somewhat less frequent. A bimodal pattern of distribution of patients with membranous nephropathy depending on the age at the beginning of the disease indicating possibly the change of etiological factors in the age groups was established. Minimal alterations, focal-segmentary glomerulosclerosis, membranous nephropathy manifested most frequently by nephrotic syndrome or subnephrotic proteinuria. Mesangioproliferative and mesangiomembranous glomerulonephritis manifested by nephrotic, proteinuric-hematuric syndrome and were the main cause of the PGN hematuric form. The highest incidence of pronounced tubulointerstitial changes in mesangiocapillary and diffuse fibroplastic glomerulonephritis is noted this explaining a considerable lowering of the kidney function in these two forms of PGN.


[Color television system for the microscope].

The characteristics of colour TV system made on the basis of microscope MBI-15 with the use of imported details are presented. The system can be used for clinico-anatomical conferences, scientific lectures, in teaching, and allows to avoid laborious preparing of colour slides. The design made it possible to keep information (images) in form of figures (for example, by means of a personal computer) and video-cassettes. This may be used in computer morphometry and quantitative analysis in histochemistry.


[Mechanisms of arrhythmias and heart fibrillation in myocardial ischemia and the possible role of Purkinje cells in their origin].

Morphological, metabolic and functional features of the heart subendocardial areas under normal conditions and at early periods of myocardial ischemia are considered in this review. It is suggested that the arrhythmias and fibrillation developing at early stages of ischemia arise in the subendocardium. The initiators of this process may be altered Purkinje's cells as precisely these cells in ischemia acquire certain characteristics necessary for the development of ectopic foci, namely: cells remaining viable should have a lowered threshold of excitability and a capacity for automatism. Further study of Purkinje's cells is particularly important for understanding of causes and prevention of a high lethality at acute phase of the myocardial infarction.


[Morphology of hairy-cell leukemia].

Trepanobiopsies from 62 patients are studied and histological features of hairy-cell leukemia in the bone marrow are characterized; the criteria of its differentiation from other lymphoproliferative diseases are given. Morphological changes in the spleen, liver, lymph nodes are described on the basis of clinical and post-mortem data. An important role of trepanobiopsy in hairy-cell leukemia diagnosis is shown.


[Alterations of nucleoli and nucleolar organizers in proliferative cystitis and transitional cell carcinoma of the urinary bladder].

The values of the nucleolar and granular rates as well as the number of Ag-granules were increased during the process of malignancy of transitional epithelium in proliferative chronic cystitis and transitional cell bladder carcinoma, including invasive tumours at both ultrastructural and light microscopic levels. The results may serve as additional criteria in the assessment of the risk of bladder mucosa malignancy and prognosis of the transitional cell bladder carcinoma.


[Morphofunctional basis for disturbances of transport-trophic myocardial function in hypercholesterolemia].

Adaptive and pathologic changes limiting the myocardial circulation and changing the microvessel circulation are described in rabbits with alimentary hypercholesterolemia. The most important changes were the sludge-syndrome, multifactorial endotheliocyte deformity and capillary compression in the perivascular oedema, sclerosis and cardiomyocyte damage. Dyslipoproteinemia is shown to initiate the circulatory hypoxia and diffuse cardiosclerosis beyond the direct connection with atherosclerotic damage of the main heart arteries.


[Age-related dynamics of aortic atherosclerosis of various ethnic groups of the male population of Andizhan].

Quantitative study of the aortic atherosclerosis was performed in 587 males of the indigenous (IP) and in 149 males of non-indigenous population (NIP) at the age of 20 to 59 years in the city of Andizhan according to the WHO program. The course of atherosclerosis was more severe in NIP persons although at the age of up to 40 years the differences in the mean surface of atherosclerotic changes were lacking. The surface of lipid spots was similar in both groups or was somewhat lower in NIP, but these spots in NIP persons were thicker and contained more foam cells and more areas of diffuse lipid deposition. The rate of the surface increase of prominent lesions drastically increased in the 5th decade of life in both groups. The frequency of rhythmical structures in the aorta was the same in both groups; their incidence was higher in cases of pronounced atherosclerosis.


[Functional state of the adrenals in various forms of arterial hypertension].

The study of structural changes and changes of the aldosterone content (AC) in the surgically removed adrenals of patients with different clinical variants of combination of the arterial hypertension (AH), low--renin hyperaldosteronism and space--occupying lesions in the adrenals found by CT was carried out. In 15 of 20 patients after adrenalectomy the diagnosis of the primary aldosteronism (PA) was established, in 4 cases diagnosis of the hypertension, 2B degree, and in one case the diagnosis of Cushing disease. The functional state was evaluated according to AC in the adenomas and macronodes and in the adjacent cortex as well as by nuclei size of cells producing aldosterone. The aldosterone hyperproduction was shown to be associated with local adenoma in some cases and with hyperactive cortex in the others this being reflected in the course of AH and in the adrenalectomy hypotensive effect.


[Prevalence of atherosclerosis of the aorta and major arteries in female inhabitants of Tashkent].

Atherosclerosis of the aorta and of practically all major arteries, including internal iliac arteries in females is studied for the first time. Visual-planimetric evaluation of the atherosclerosis degree of the arteries stained with Sudan IV in Tashkent females 20-89 year-old with regard to the age, ethnic and cause of death is performed. The highest incidence was observed in the age period of 40 to 59 years. Indices of the atherosclerotic damage considerably differed in various arterial regions being higher in males and in non-indigenous population. More severe course of atherosclerosis in Tashkent females was noted in the last decades.


[Status and prospects for the development of a pediatric pathology service in the Russian Federation].

The paper presents the main points of the reports made at the constituent conference of the Russian Society of Pathologists. Some urgent measures for improving this service are suggested including the creation of independent pediatric bureaus of pathology and some others.


[Status and prospects for the development of a pathology service in the Russian Federation].

General crisis in the country exerts a negative influence on the pathology service in Russian Federation. There is a lack of pathologists--only 58.6% of the vacancies were filled in 1989. About 17% of the buildings are in need of emergency repairs, 30% require major repairs. The supply of microscopes, instruments, reagents, etc., is unsatisfactory. The number of autopsies in the hospitals decreased by 19.4% in 1989 compared to 1986. Centralization of the pathology service is a positive element, and 22 bureaus of pathology functioned in Russian Federation in 1989. Transition to the market economy will place new problems concerning skilled staff, modern equipment and finances.


[Progression of tumors: etiologic, morphologic and molecular-biological aspects].

Two aspects can be distinguished in multistage carcinogenesis: etiological one (every stage is induced by a specific for this stage agent) and morphobiological aspect (every stage is characterized by specific morphological, genetic and other properties). The schema of the multistage carcinogenesis is presented in which morphological stages (diffuse and focal hyperplasia, benign tumours, dysplasia, carcinoma in situ, various phases of malignant tumour progression) are placed against genetic alterations. L. Foulds concept of tumour progression is discussed with special emphasis on precancerous stages, possibilities of cancer development de novo, and independent progression of different tumour characters. The following types of carcinogenesis are listed on the basis of interrelationship between etiological and genetic factors: 1) carcinogenesis induced by genotoxic agents; a) one agent is acting at high dose and for a long time thus ensuring the activation of protooncogenes and all stages of tumour progression (initiation, promotion, various phases of malignant tumour); b) those acting during a very short time, however sufficient for developing the genetic program working automatically without further exposure to known carcinogens (irradiation in case of the atomic bomb explosion or effect of short-living alkylating agents): in this case there is no stage of promotion; 2) carcinogenesis by non-genotoxic carcinogens (their mode of action is still unclear, the only human example is carcinogenesis by hormones); 3) development of tumours in frane of the two (or three) stage carcinogenesis when every stage is provoked by its own etiological factor, no human examples are known as yet; 4) development of tumours due to the genetic mechanism making the organism highly susceptible to the minimal doses of carcinogens as is the case with skin cancer by ultraviolet light in patients with xeroderma pigmentosum, the genetic damage in itself has nothing to do with tumour formation; 5) genetic damage leading to the development of tumour without visible participation of any known carcinogens or promoters (gene Rb in retinoblastoma, gene Wt in Wilms tumour, etc.).


Program design for adult learners.

The voice of adult learners is often overlooked in the design of programs that are specifically constructed to meet their learning needs. This article portrays the design of a midwifery re-registration program that commenced with a survey of non-practising midwives. The organisation of the course, together with the processes involved in curriculum building are explained. The adult learner, particularly in a situation where previous knowledge, skills and experience are vital inputs to the design of programs, have been highlighted.


My room--not theirs! A case study of music during childbirth.

The more holistic goals of midwifery are emphasised in this paper through the therapeutic application of music. Material resulting from an interview with a mother after the birth of her first child is used to demonstrate many of the psychological and socio-psychological benefits of listening to music during labour. Several of these benefits were experienced not only by the mother-to-be but also by her partner and the hospital staff. In the outlining of numerous advantages for employing music listening in birthing situations--in particular, comfort and security, self-esteem, socialisation and personal control--the importance of music as a nursing intervention is stressed.


An in vitro study of polymorphonuclear leucocyte-mediated injury to human gingival keratinocytes by periodontopathic bacterial extracts.

Human gingival keratinocytes were cultured and, after the first passage, subjected to cell detachment assays with polymorphonuclear leucocytes (PMNs) and/or sonic extracts from Actinobacillus actinomycetemcomitans Y4, and Eikenella corrodens 1073. The effector-to-target cell ratio was 30:1. Bacterial extracts alone caused no disruption of keratinocyte monolayers. PMNs alone also caused only minimal detachment after 14 h incubation. Adding A. actinomycetemcomitans to the PMN-keratinocyte co-cultures at the concentration of 100 micrograms/ml caused dramatic cell detachment. The effect of A. actinomycetemcomitans was heat labile and not inhibited by polymyxin B. Cell detachment was inhibited by alpha 1-antitrypsin, whereas catalase and superoxide dismutase could not prevent it. No lysis of keratinocytes was observed after incubation, as judged by 51Cr release. E. corrodens had little effect even at the concentration of 1000 micrograms/ml. H2O2 and partially purified PMN elastase also caused detachment of keratinocytes. These data indicate that PMNs can cause non-lytic detachment of keratinocytes when interacting with certain bacteria.


Activation of sympathetic fibres in the pulp by electrical stimulation of rat incisor teeth.

Previous studies of the sympathetic nerve supply of the rat mandibular incisor pulp have shown conflicting results. Here, the neurovascular control of the rat lower incisor pulp was investigated by stimulating the tooth crown and the cervical sympathetic trunk electrically and monitoring blood-flow changes in the pulp by laser Doppler flowmetry. In addition the presence of noradrenaline (NA) in the pulp and gingiva was examined biochemically in untreated and sympathectomized animals by high-performance liquid chromatography. The tissue concentrations of NA in the pulp were 11-fold greater than those of gingiva. Surgical sympathectomy significantly reduced the NA content in the pulp by 76%. Monopolar electrical stimulation of teeth (25-50 microA) for 1 min resulted in a frequency-dependent reduction followed by an increase in pulpal blood flow. At 16 Hz the reduction in blood flow was 65% and the subsequent increase was 9%. After intravenous administration of the alpha-adrenoceptor antagonist phentolamine, the stimulation-induced reduction in pulpal blood flow was diminished by 94% while the increase was significantly enhanced (from 9 to 40%). Infusion of the beta-adrenoceptor antagonists propranolol and timolol significantly reduced this increase in blood flow by 53 and 55%, respectively. Preganglionic sympathetic nerve stimulation also induced a frequency-dependent reduction followed by a slight increase in pulpal blood flow. This reduction in blood flow was almost abolished after alpha-adrenergic blockade and there was no subsequent increase. These findings suggest that there are sympathetic nerve fibres in rat incisor pulp and that they are activated upon monopolar electrical stimulation of teeth resulting in alpha- and beta-adrenoceptor mediated blood-flow responses.


Association between the expression of murine 72 kDa type IV collagenase by odontoblasts and basement membrane degradation during mouse tooth development.

In situ hybridization was used to study the expression of the 72 kDa type IV collagenase gene and its association with morphogenesis and cell differentiation during advancing mouse tooth development. The epithelia were completely negative during all developmental stages. The dental mesenchyme was uniformly positive during the early stages of tooth morphogenesis, and no association of type IV collagenase with morphogenetic events was observed. However, at the bell stage the expression increased in differentiating preodontoblasts. Expression was intense in the odontoblasts during secretion of the first predentine matrix. The expression was, however, transient; it decreased around the time when mineralization of dentine started until it completely ceased. Transcripts for 72 kDa type IV collagenase also gradually disappeared from the dental pulp. The expression of 72 kDa type IV collagenase was also strong in the osteoblastic cell lineage. The preosteoblasts at the beginning of the formation of mandibular bone as well as the osteoblasts of the alveolar bone expressed more 72 kDa type IV collagenase than did other mesenchymal cells. The increased gene expression in the odontoblasts correlates with the disappearance of the dental basement membrane as shown by immunolabelling with antibodies against type IV collagen. The onset of increased expression in the odontoblasts preceded the disappearance of the basement membrane and at the time when type IV collagenase transcripts were lost from all odontoblasts the basement membrane was completely removed. It can be speculated that during early stages of tooth development the 72 kDa type IV collagenase acts as a gelatinase whereas during later stages, when odontoblasts and ameloblasts differentiate and the deposition of predentine and enamel matrix is initiated, the enzyme may act as a type IV collagenase and contribute to the degradation of the dental basement membrane.


The effect of periodontal disease on sulphated glycosylaminoglycan distribution in the sheep periodontium.

The glycosylaminoglycan (GAG) distribution in the soft periodontal tissues of the sheep was investigated topographically in healthy tissue and the changes associated with periodontal disease determined and correlated with increasing disease severity. Total sulphated GAG content was determined spectrophotometrically and the proportions of individual GAGS measured after separation by cellulose acetate electrophoresis. In healthy tissue, total sulphated GAG distribution was found to mirror that described previously for collagen, being highest in the gum-pad region and adjacent to alveolar bone. This changed markedly in relation to periodontal disease. Early disease-related changes in GAGs were first detected adjacent to alveolar bone. In severely diseases tissue, an overall increase in sulphated GAG, particularly in the gum-pad region, was present. Increasing disease severity was accompanied by a decrease in dermatan sulphate and a concomitant increase in the proportion of chondroitin-4-sulphate. These changes may be due to the combined effects of chronic inflammation and alterations to the mechanical loading of the tissue owing to loss of attachment.


Dental morphology of 45,XO human females: molar cusp area, volume, shape and linear measurements.

A three-dimensional analysis of the maxillary first molars of five 45,XO females and comparison with normal females and males demonstrated a reduction of cusp areas and volumes in the 45,XO females. Mesiodistal and faciolingual dimensions were also reduced. The reductions in basal area and volume were greater in the later developing, distal cusps. In normal females the second X chromosome apparently does not exert an effect on the cusp height but increases the basal area. The results further indicate that the Y chromosome in normal males increases both cusp height and basal area. The steepest cusps were found in 45,XO females while the shallowest were in the control males. Both sex chromosomes thus seem to affect the shape of the cusp but the resulting phenotypes differ.


Comparison of phosphoprotein isolated from mature and immature human tooth roots.

Mature (average patient age = 29.5 yr, closed apical foramen) and immature (average patient age = 17.5 yr, open apical foramen) root shards were placed in dialysis tubing and demineralized to completion using either 10% disodium EDTA plus protease inhibitors or 0.6 N HCl. The demineralized shards were re-extracted (five times) with 0.05 M tris-HCl, 1.0 M NaCl and then collagenase digested. No major differences were observed in chromatograms of extracts, re-extracts or collagenase digests from root shards demineralized in either way. In contrast, chromatograms of immature and mature roots showed qualitative differences. Chromatograms of mature roots demineralized in either way showed broader protein peaks and less organic phosphorus than those from immature tooth roots. A distinct band amid degraded phosphoprotein (150 K) was found in SDS-PAGE gels (7.5%) from EDTA-extracted immature tooth roots but not from mature tooth roots. Electroelution of this band revealed a typical phosphoprotein amino-acid profile containing increased aspartic acid and serine residues. Comparison of the total phosphoprotein and amino acid composition of extracts, re-extracts and collagenase digests revealed that phosphoprotein, serine and to a lesser extent aspartic acid were recovered in greater quantities from immature roots than mature tooth roots. These data suggest that the degree of maturation is crucial to the isolation of an intact phosphoprotein and provides additional evidence that human dentine phosphoprotein undergoes amino acid compositional changes during maturation.


Effects of clonidine on the calcium content and morphology of rat salivary glands.

These effects were examined with and without pretreatment of animals with reserpine and the adrenergic antagonists prazosin (alpha 1), yohimbine (alpha 2) and propranolol (beta). The effects of clonidine on glandular concentrations of norepinephrine and dopamine also were examined. These effects were compared with those of xylazine, a presynaptic alpha 2-adrenergic agonist. A single, high dose of clonidine followed by an overnight fast caused marked increases in calcium content and acinar secretory granules in the submandibular and sublingual glands, similar to those caused by reserpine. However, the calcium content of the parotid gland was not altered by clonidine, although there seemed to be a modest increase in acinar secretory granules. The clonidine-induced increase in submandibular calcium content could not be attributed to any adrenergic receptor activity since it was not blocked by either alpha- or beta-adrenergic antagonists. Unlike reserpine, clonidine did not affect catecholamine concentrations in the parotid and submandibular glands. Pretreatment with reserpine did not significantly alter the clonidine-induced increase in submandibular calcium content. It is likely that the greater accumulation of acinar secretory granules is related to the increased calcium stores of the glands in clonidine- and/or reserpine-treated rats. The large differences in calcium content among the three glands might be attributable, in part, to differences in the calcium-binding capacity of their secretory granules. Possible mechanisms for the clonidine effects on salivary-gland calcium include disturbances in membrane-associated pools or gating mechanisms for calcium, which need further study.


Microfibrillar components in dental pulp: presence of both type VI collagen- and fibrillin-containing microfibrils.

Microfibrillar elements were isolated from developing and formed bovine dental pulp by a procedure involving bacterial collagenase tissue digestion and chromatography on Sepharose CL-2B. Two microfibrillar assemblies could be demonstrated. Type VI collagen microfibrils with a characteristic periodicity of about 100 nm appeared as long, thin, flexible filaments. In a number of cases these structures aggregated by lateral association. Microfibrils of 10-14 nm dia were identified as containing fibrillin on the basis of their distinctive, periodic, beaded morphology. In addition to long, single strands there were instances of chains coalescing to give amorphous aggregates. No differences in the type of microfibrillar assemblies were evident between developing and formed pulp, although fibrillin-containing microfibrils were more abundant in formed pulp.


Quantitative electron-microscopic analyses of pulpal nerve fibres in the mouse lower incisor after neonatal capsaicin treatment.

A single dose of capsaicin (50 mg/kg) was injected subcutaneously into four mice on day 2 of life; four untreated mice were used as controls. Six months later, a drop of 30 microM capsaicin was instilled on to the cornea of all the mice and the number of times the eyes were wiped was counted to assess the effect of capsaicin on trigeminal sensory neurones. Ultrathin cross-sections were made of the apical pulp of the incisors on both sides of control (n = 8) and capsaicin-treated animals (n = 8). Electron micrographs of pulp nerves were taken and enlarged to a final magnification of x34,000. The numbers of unmyelinated axons in the pulps of all 16 incisors and of unmyelinated axons per Schwann cell in the pulps of four incisors each from the control and capsaicin-treated groups were counted. The short diameters of unmyelinated axons were measured with a computer-operated image analyser. The number of eye wipings was eight-fold less in the capsaicin-treated than in the normal group. This finding clearly indicated that capsaicin irreversibly affected the chemogenic nociceptive trigeminal neurones. The mean number of unmyelinated axons was 345 in controls and 217 (37.1% reduction) in capsaicin-treated animals. The number of unmyelinated axons of less than 0.6 microns dia was 41.5% less in capsaicin-treated mice than in controls. Thus, fine unmyelinated axons in the mouse incisor pulp are capsaicin sensitive, and they are assumed to be nociceptive fibres conveying pain stimuli from the tooth. Capsaicin affected Schwann cells, even those with few unmyelinated axons.(ABSTRACT TRUNCATED AT 250 WORDS)


Occurrence of a postpermanent mandibular fourth premolar in a moose bull (Alces alces L.).

By a number of diagnostic criteria a supplemental premolar in the right hemimandible of a 6-yr-old moose bull from Bielorussia was identified as a postpermanent fourth premolar. Its development, which must have started at least 3 yr after the permanent fourth premolar had reached its functional position, resulted in rotation of the P4 by 80 degrees, as well as extensive resorption on both root and crown.


Characterization of amelogenin mRNA from secretory- and maturation-stage rat incisor enamel.

Amelogenins are extracellular matrix proteins expressed at the secretory and the maturation stages of enamel formation. To characterize amelogenin transcripts from these two stages, rat incisor enamel-organ mRNA was hybridized to a mouse amelogenin cDNA. Northern analysis of the mRNA showed four amelogenin transcripts at the secretory stage. Two of these were larger than previously described, and would code for high molecular-weight amelogenins (> 40 kDa). These transcripts were similar in size to at least three bovine amelogenin mRNAs. At the maturation stage, one transcript slightly smaller than the major amelogenin transcript from the secretory stage was apparent. These differences may indicate a selective splicing of amelogenin mRNA in the maturation stage as compared to the secretory stage of enamel development.


Bite-force development, metabolic and circulatory response to electrical stimulation in the canine and porcine masseter muscles.

The development of fatigue was investigated by electrical stimulation in 15 domestic pigs (1 yr old, 70-90 kg body weight) and seven adult dogs (3 yr old, 45 kg body weight). After anaesthesia, silver electrodes were implanted in the anterior and posterior parts of the right masseter muscles. The contralateral muscle was used as control. The bite force was measured. Muscle biopsies were obtained from the anterior, central and posterior parts, and were immediately frozen in liquid nitrogen. A fluorometrical analysis by enzymatic methods for glycogen, glucose, creatine phosphate, NAD, NADH, lactate and pyruvate was made. Blood flow was measured by 133Xe wash-out; oxygen consumption was monitored with an oxygen electrode. The porcine masseter was continuously stimulated for 60 min (100 V, 4 Hz and 2 ms). The canine masseter was intermittently stimulated (100 V, 20 Hz and 2 ms). The contraction was repeated four times, with a 10-min rest between. The porcine masseter could sustain longer endurance times than the canine masseter, which was easily fatigued. A marked substrate depletion was evident. The precontractional contents of glycogen, glucose and creatine phosphate were reduced. Lactate accumulation was evident (2-4 times more in the porcine and 4-8 times more in the canine masseter). The NADH concentration increased and the NAD content decreased. Blood-flow impairment (80% reduction in the dog, 60% in the pig) was observed. After the contraction phase, there was a hyperaemia (58% elevation of blood flow in the pig masseter, 45% in the canine). The oxygen tension followed in magnitude and time the blood-flow changes. These circulatory variables returned to normal after recovery. The high degree of substrate depletion, blood-flow impairment and a simultaneous decrease in oxygen transport to the contracted muscle, in combination with a prominent lactate accumulation, may induce a decrease in bite-force production.


The effect of density on the 10MV photon beam penumbra.

An investigation into the density dependence of the penumbra of the Varian Clinac 18/10 10MV photon beam has been carried out. A water/lung phantom was constructed of polystyrene (r = 1.04 g cm-3) and cork (r = 0.23 g cm-3), in which interfaces exist both parallel and perpendicular to the beam axis. The irradiation of the phantom was also simulated using the EGS4 Monte Carlo system with a cartesian voxel geometry. Experimental (TLD) and Monte Carlo dose profiles are in close agreement, and show a large degree of penumbral broadening in the lung region. This broadening is due primarily to lateral electronic disequilibrium occurring at a larger distance from the geometric beam edge in lung than in water. This disequilibrium can also cause the dose in lung to drop below the dose in water at the same depth and off axis distance, even though the radiological depth is less in lung. Monte Carlo simulations were also performed where the dose is separated into primary and scattered components, for homogeneous media of densities 0.25, 0.50, 0.75 and 1.00 g cm-3. The penumbral width of the primary dose profile was found to be almost constant with depth for a point source of photons (after the initial build-up region), where the lateral distances from the 95-50% and 50-5% dose levels on the dose profile (normalised to the dose at the central axis) are equal in all cases. Also, primary penumbra width was found to be almost inversely proportional to density. The primary penumbra for a unit density material can be fitted accurately by an exponential forming function with empirical determined coefficients. The penumbral shape for the lower densities can then be closely fitted by scaling the coefficients in proportion to density. This scaling method has application in treatment planning, where the predicted primary penumbra shape should take account of inhomogeneities, and is particularly important in matching adjacent fields. When the scattered dose component is added to give the total dose, penumbral width increases because the scattered dose penumbra is wider than that of the primary dose. Also, the inverse proportionality of the penumbra width with density does not hold for the scattered dose. The relative contribution of the scattered dose increases with density. Therefore, the inverse proportionality of penumbra width with density does not hold for the total dose.


A flexible automated data acquisition system for ophthalmic electrophysiology.

An automated system for performing VER, ERG and EOG measurements has been developed and is now in clinical use. Considerable effort has been made to retain the flexibility of non-automated systems while providing an easily followed procedure for standard tests which can be undertaken by staff with minimal training. Clinical experience has shown that the standard parameters for data acquisition which are optimal for normal subjects are not always suitable for patients who are uncooperative or have poor vision. These parameters are under software control and can be amended interactively if necessary. The five main functions of the software, namely data acquisition, data selection and commentary, report generation, patient database operation and waveform archiving are written as separate modules and called as required. During data acquisition the software rejects gross artefacts and in the case of EOG's warns of patient non-cooperation. After an ERG or VER has been acquired the operator identifies features in the waveforms and the computer measures them whereas for EOG's the measurement process has been automated. When the waveforms are subsequently reviewed by the operator they are presented in a logical sequence, a sequence which does not depend on the order in which they were acquired, and this sequence is maintained when they are printed out later in the completely automated report phase.


An efficient algorithm for MR image reconstruction and compression.

In magnetic resonance imaging (MRI), the original data are sampled in the spatial frequency domain. The sampled data thus constitute a set of discrete Fourier transform (DFT) coefficients. The image is usually reconstructed by taking inverse DFT. The image data may then be efficiently compressed using the discrete cosine transform (DCT). We present here a method of using DCT to treat the sampled data, which combines two procedures, image reconstruction and data compression. This method may be particularly useful in medical picture archiving and communication systems (PACS) where both image reconstruction and compression are important issues.


Radiotherapy x-ray dose distribution beyond air cavities.

Radiation oncologists are particularly concerned about tumours growing on the surface of air cavities in the head and neck regions, which involve treatment with small x-ray fields. An inhomogeneous dose distribution exists within and beyond the cavity. This is caused by the loss of electron equilibrium and the attenuation of both the primary and scattered photons is altered. The scatter function photon beam models for tissue inhomogeneity, such as the ETAR correction algorithm, currently implemented in commercial treatment planning systems do not predict the dose distribution accurately in many situations where lateral electron equilibrium does not exist. Using a Markus ionization chamber and different solid water slabs to simulate different air cavities, it is found that internal body cavities, depending upon their sizes, experience underdose or overdose on the distal surfaces of the cavities when compared with the results predicted by an ETAR correction method for 6 MV and 18 MV x-ray beams. For an infinitely long air passage of dimensions 2 cm x 2 cm, the error in the ETAR correction for a 6 MV x-ray beam is 4.8%, 0.5% and 1.1% for the field size of 5 cm x 5 cm, 7 cm x 7 cm and 10 cm x 10 cm respectively. The ETAR correction is accurate to within 1.6% for a 6 MV x-ray beam provided that the field size is 5 cm across the cavity and greater than 7 cm along it.


Survey and risk assessment of radium contamination in a historic desk.

For many years the Physics Department of The University of Adelaide has owned a fine cedar desk which dates from before 1900. It was used by professors, including Sir Kerr Grant, for many years and probably Sir William Bragg. In April 1991, approximately 80 kBq of radium contamination was found in parts of the desk. Because of concern for the health of those who had used the desk, a thorough study was made of the nature and extent of the contamination and the risks to health from radon and from ingesting some of the radioactive material from contact with articles such as pencils, stamps and envelopes kept in the drawer. The risks have been shown to be small compared to other acceptable risks in modern society. The desk has been decontaminated and restored.


[Intravenous infusion of volatile fatty acids as a metabolic loading test for the evaluation of the possible effects of butafosfane on the energy metabolism of cattle].

In 10 German Black and White heifers metabolic stress reactions were induced by intravenous infusion of propionate and butyrate to demonstrate possible effects of prior medication with butafosfane. This investigation included a preliminary feeding period, followed by 18 hours of feed withdrawal, premedication with butafosfane or placebo and infusion of volatile fatty acids. Reactions were assessed by monitoring biochemical and clinical parameters. A total of 19 blood samples per series were taken in which the concentrations of glucose, free fatty acids, volatile fatty acids, beta-hydroxybutyrate, cortisol and insulin were measured. Clinical parameters recorded included heart rate and respiratory rate. During loading with propionate effects of butafosfane on plasma levels of cortisol and insulin could be demonstrated. The differences noted in the other parameters were only insignificant. The loading test chosen is suitable for the demonstration of effects of butafosfane on metabolic parameters which could hardly be influenced by this substance in unstressed animals.


[Natural behavior--a natural concept?].

Animal behaviour is a function of both endogenous and exogenous stimuli. To survive, the animal registers, predicts and tries to control its environment. This leads to a certain behaviour pattern. A disturbed behaviour and a state of deteriorated health is the consequence if the behaviour pattern is obstructed.


[Seroepidemiological studies of the detection of leptospires of the sejroe group in cattle in middle Thuringia].

From 1983 to 1989 14,361 head of cattle from the middle Thuringia region were investigated by means of microagglutination test for the presence of Leptospira serovars hardjo, saxkoebing and sejroe. 4,484 samples from cattle with abortion (1983-1985) and a random sample of 5,284 cattle sera (1985) were investigated giving priority to L. hardjo. Furthermore 3,293 samples from cattle with abortion (1986/87) and a random samples test of 1,300 cattle sera (1989) were tested giving priority to L. saxkoebing. 2.5 percent of the cows having aborted and 10.3 percent of cattle tested out of the whole livestock showed antibodies against L. hardjo. The test for presence of L. saxkoebing demonstrated in cattle with abortion a seroprevalence of 14 percent and in the random samples tests a portion of reagents of 11.3 percent. The degree of infection differed regionally, the highest degree was found in livestocks of the northern part of the Thuringia forest. The prevalence of L. saxkoebing in cattle with abortion from large dairy farms was 18.6 percent, which is higher than the average in the region examined. Antibodies against hardjo and sejroe were interpreted as caused by cross reactivity.


[Possibilities for the establishment of pneumonia-free swine herds by immunization and administration of cefquinome].

The application of Bordetella and Pasteurella inactivated and adsorbed vaccines together with cefquinome to sows, piglets and weaners led to a significant reduction of the incidence of rhinitis atrophicans and pneumonia. The frequency of positive isolates of P. multocida, H. parasuis and A. pleuropneumoniae out of nasal swabs was reduced during the treatment.


[The effect of Eperythrozoon suis infection on the osmotic fragility of erythrocytes].

Osmotic fragility of erythrocytes was tested in weaned pigs experimentally infected with Eperythrozoon (E.) suis. Acute eperythrozoonosis of splenectomized pigs led to an increase of osmotic fragility. It is supposed that E. suis infection causes a structural change in erythrocyte membrane. Possible mechanisms of this cell membrane injury are discussed.


[The selection of Salmonella gallinarum, Salmonella pullorum and Escherichia coli in chickens after chloramphenicol administration].

The effect of a chloramphenicol administration was examined on the selection of E. coli of the chicken intestinal flora, and of the infectious S. gallinarum and S. pullorum strains. On the other hand, an effort was made to detect the frequency of the resistance transmission of E. coli to above mentioned sensitive salmonella strains. Fourteen chicken, 12 infected and 2 negative controls were used. It was found that the enteric E. coli strains became resistant in a week's time. Besides, the strains that were used for infecting the chicken neither were selected through the chloramphenicol administered nor did they take the E. coli resistance, via R-factor transmission.


Cell-shape-associated transcriptional activation of the p52(PAI-1) gene in rat kidney cells.

The microfilament-disrupting agent cytochalasin D (CD) increased (by 10-22-fold) the synthesis de novo and extracellular matrix deposition of plasminogen-activator inhibitor type-1 [p52(PAI-1)] in normal rat kidney (NRK) cells. Transition from a flat to a round phenotype occurred concomitantly with, and may actually precede, p52(PAI-1) induction; both the morphological and p52(PAI-1) responses were dose-dependent. Augmented synthesis became evident between 4 and 5 h of treatment of NRK cells with 100 microM-CD, correlating with a transition from 25 to more than 60% rounded cells. CD-associated increases in p52(PAI-1) mRNA abundance and protein biosynthesis were maximal between 6 and 8 h of continuous CD exposure, declined by 50% thereafter, but remained elevated (by at least 6-21-fold respectively over control values) for 24 h. Changes in p52(PAI-1) mRNA abundance at this 24 h point reflected an approx. 5-fold increase in p52(PAI-1)-gene transcription. These data confirm previous suggestions, based on actinomycin D-sensitivity of the inductive response [Higgins & Ryan (1992) Biochem. J. 284, 433-439], that CD-mediated increases in p52(PAI-1) expression are at least partly due to transcription-level events. Since CD also augments specific cellular responses to growth factors or cytokines, the potential effectiveness of this inducer was evaluated both in the presence and absence of serum growth factors using quiescent NRK cells [a growth state in which p52(PAI-1) is not expressed] as a model system. Induction of p52(PAI-1) synthesis and matrix deposition in CD-stimulated quiescent NRK cells was as efficient under growth-factor-deficient conditions as when CD was added simultaneously with serum. CD alone is thus a complete inducer of p52(PAI-1) expression in NRK cells, an observation that supports the contention that cell shape is an important regulatory element in p52(PAI-1)-gene control.


Isolation and characterization of different C-terminal fragments of dystrophin expressed in Escherichia coli.

Dystrophin, the protein product of the Duchenne muscular dystrophy gene, is thought to belong to a family of membrane cytoskeletal proteins. Based on its deduced amino-acid sequence, it is postulated to have several distinct structural domains; an N-terminal region; a central, rod-shaped, domain; and a C-terminal domain [Koenig, Monaco & Kunkel (1988) Cell 53, 219-228]. The C-terminal domain is further divided into two regions; the first has some sequence similarity to slime mould alpha-actinin, and is rich in cysteine residues; this is followed by the C-terminal amino-acid sequence that is unique to dystrophin. Dystrophin is very difficult to purify in quantities sufficient for detailed studies of the structure/function relationships within the molecule. Therefore, in this study, we have expressed selected fragments of the C-terminal region of dystrophin, as fusion proteins, in Escherichia coli. Importantly, we describe the first successful purification, from E. coli lysates, of large quantities of fragments of dystrophin in a soluble form. The first fragment, termed CT-1, encodes the C-terminal 201 amino acids of the protein; the second, termed CT-2, spans the cysteine-rich region of the C-terminal domain. These fusion proteins were identified by their mobility in SDS/PAGE, by their interaction with appropriate affinity columns and by their reactivity with anti-dystrophin antibodies. The fragment CT-2, which spans a region containing putative EF-hand-like sequences, was found to bind Ca2+ in 45Ca2+ overlay experiments. In addition, we have discovered that the fragment CT-1, but not fragment CT-2, interacts specifically with the E. coli DnaK gene product [analogue of heat shock protein 70 (hsp70)]. This interaction is disrupted, in vitro, by the addition of ATP. Our results indicate that the two C-terminal fragments of dystrophin have differing biophysical properties, indicating that they may play distinct roles in the function of the protein.


Site-directed mutagenesis and substrate-induced inactivation of beta-lactamase I.

The substrate-induced inactivation of beta-lactamase I from Bacillus cereus 569/H has been studied. Both the wild-type enzyme and mutants have been used. The kinetics follow a branched pathway of the type recently analysed [Waley (1991) Biochem. J. 279, 87-94]. The substrate cloxacillin (a penicillin) formed an acyl-enzyme (characterized by m.s.), and it was probably the instability of this intermediate that brought about inactivation. A disulphide bond was introduced into beta-lactamase I (the wild-type enzyme lacks this bond) by site-directed mutagenesis: Ala-77 and Ala-123 were replaced by cysteine. Spontaneous oxidation yielded the disulphide. The activity of this newly cross-linked enzyme was a little diminished, but the stability towards inactivation by cloxacillin was not increased. A second mutant of beta-lactamase I was studied: this mutant lacked the first 17 residues, i.e. the first alpha-helix. The mutant had reduced activity towards ordinary (non-inactivating) substrates and no hydrolysis of cloxacillin could be detected. These mutant enzymes were expressed in Bacillus subtilis, and were purified from the extracellular medium.


Sheep 6-phosphogluconate dehydrogenase. Revised protein sequence based upon the sequences of cDNA clones obtained with the polymerase chain reaction.

Sheep liver 6-phosphogluconate dehydrogenase (6-PGDH) is an enzyme of the pentose phosphate pathway. Evidence has appeared which suggests that the 6-PGDH protein sequence determined previously by direct analysis of the protein isolated from ovine liver is incorrect. Determining the enzyme's DNA sequence was considered to be the best way of solving the problem. In the first instance, a degenerate forward and a degenerate reverse primer were designed on the basis of the known protein sequence, and a partial-length cDNA clone was isolated from total sheep liver cDNA using the polymerase chain reaction. The clone encoded the expected part of the protein sequence. The clone was unsuccessfully used as a prime-cut probe to screen a sheep liver library and a bovine heart library. As a result, the polymerase chain reaction was utilized again to successfully generate a family of overlapping cDNA clones encoding a mature protein of 482 amino acids. The mature protein sequence encoded by the cDNA differs significantly from the sequence derived by direct analysis of the protein, but on closer examination the fundamental difference is caused by the incorrect placement of three enzyme fragments obtained by cyanogen bromide cleavage during the direct sequence analysis of the protein. Placing the fragments in the correct order results in the two sequences being virtually identical except for some minor amino acid changes between the amide and acid forms, and a small number of deletions and insertions.