[Use of modern methods in prenatal diagnosis of congenital adrenal hyperplasia--examination protocol].

The prenatal diagnosis of congenital adrenal hyperplasia (CAH) in the second trimester of gestation (as done so far in this country) is late and unsuitable with regard to possible interruption of pregnancy. Modern methods of early prenatal diagnosis are based on examination of material from chorionic villi (6th-10th week of gestation) or early amniocentesis (10th-12th week of gestation). The authors present their decision taking scheme, examination protocol and their own experience with the use of molecular genetic analysis and assessment of 17-hydroxyprogesterone from early amniocentesis in the prenatal diagnosis of CAH. This procedure is a priority in this country.


[A non-classic form of congenital adrenal hyperplasia].

The authors examined five children with the non-classical form of adrenal hyperplasia. The clinical symptoms of the disease comprise: early congenital pubic hair, small stature, accelerated bone maturation, infertility, in girls hirsutism, acne, menstrual disorders, amenorrhoea, in boys oligospermia, acne. Laboratory examination reveals slightly elevated 17-OH progesterone values during the synactene loading test. It is an autosomal recessively hereditary disease. It is due to an allelic variant of the gene for 21 hydroxylase. The authors revealed a link with antigen B14. Even in the small group examined antigen B14 was present in 40%, although the prevalence in our population is 2.9%.


[Levels of plasma amino acids in normal neonates within the context of their postnatal adaptation].

The authors present values of plasma amino acid concentrations assessed on an automatic analyzer in a group of 12 mature neonates whose early and late post-partum adaptation and subsequent psychosomatic development followed-up to the age of 18 months was normal. The amino acids were assessed at the time of delivery, during the maximal weight loss of the infant after the third day and the last estimation was made before discharge from the maternity hospital when the body weight curves of all infants had a rising trend. None of the infants had neonatal jaundice and all were discharged fully breastfed.


[The effect of food supplementation with organically bound chromium on indicators of compensation in diabetic children and adolescents].

To the food of 33 diabetic patients typ I (in the age of about 15 years) organically bound chromium was added for 6 weeks. The influence on the metabolic compensation could not be proven, although in some individuals of this group the favorable effect of chromium can not be excluded.


[Immunization against diphtheria and tetanus in children with severe neurological disorders].

In the clinical department of the Institute for Sera and Vaccines 260 children with serious neurological diseases were immunized with Alditeana and Alteana. After immunization an undesirable reaction was recorded in 3.1% of the immunized children. Mostly recurrence or potentiation of clinical manifestations of the basic neurological disease of the child was involved. Investigation of the anamnestic data revealed several serious undesirable reactions after vaccination with Alditepera in a health centre for child patients.


[Ambroxol in the treatment of idiopathic respiratory distress syndrome].

The authors treated five premature infants with idiopathic respiratory distress syndrome with ambroxol, using its action on type II pneumocytes and thus also on surfactant formation. In all infants improvement of the clinical condition was achieved, no side-effects were observed.


[Problems in child nutrition. 2. Nutrition in preschool-age and school-age children. Nutrition in children in families with different life styles].

The authors presents a review of the main principles of enteral nutrition of preschool and school children. He draws attention to the unsuitable character of an alternative diet for growth and development of the child. The nutritional status of children on an alternative diet can be, however, supplemented and modified.


[Mild injuries of the head in 0-3-year-old children].

The authors discuss the problem of mild head injuries in children aged 0-3 years and based on data obtained by a retrospective examination of a group of 119 hospitalized children and data in the literature they submit recommendations as regards examination and follow-up of these children.


[Medullary carcinoma of the thyroid gland. Personal experience].

The authors present their experience with the diagnosis and treatment of medullary thyroid carcinoma (MTC) in a group of 175 patients in the records of the RI in Motol. From the above number 106 are alive, 69 of the patients who died were diagnosed at the time of the clinical manifestation of the disease, frequently an advanced stage of the disease. Of the living patients one third was treated by total thyroidectomy after active screening. MTC accounts under our conditions in recent years for 8.6% of all thyroid malignancies. The familial variant with autosomal dominant heredity accounts for 26.5% of the above number. Within the framework of the familial variant the most frequent finding is non-MEN MTC, only 30% MTC are part of the MEN 2A or MEN 2B syndromes. MTC in childhood belongs practically always to the familial variant. At present the authors' group comprises 13 children and adolescents under 18 years, incl. one boy with the MEN 2B syndrome who succumbed to the disease. In 8 children based on active screening total thyroidectomy was indicated, incl. 5 patients were by histological examination only hyperplasia of the C-cells, i.e. a precancerous condition, was detected. After operation repeatedly in all these patients the level of immunoreactive calcitonin is low which indicates an excellent prognosis. The authors draw attention to the importance of active screening in families of all direct relations of the patient where MTC was diagnosed. Complete removal of thyroid tissue at the time of hyperplasia of C-cells or the presence of tumourous microfoci has a marked positive effect on the subsequent fate of the patients.(ABSTRACT TRUNCATED AT 250 WORDS)


[Extended classification of diabetic retinopathy in children and adolescents].

The authors submit their own modification of the classification of diabetic retinopathy elaborated on the background of several years experience with the examination of children and adolescents with insulin dependent diabetes. The extended classification pattern makes it possible to differentiate and classify more accurately even slight incipient changes detected by chromatooophthalmoscopy. The presence of microaneurysms reflects well the degree of retinal damage in early stages. Special attention must be paid to intraretinal microvascular abnormalities (IRMA). The classification pattern is presented in a table included in the text.


[Occurrence of diabetic retinopathy in children and adolescents and its relation to the duration of diabetes and patient age].

In a retrospective investigation the authors give an account of a group of 302 children and adolescents with insulin dependent diabetes (mean age 14.2 years, mean duration of diabetes 6.4 years) whom they examine regularly by chromatoophtalmoscopy and evaluate the findings according to their own extended classification of diabetic retinopathy. In 29% of the patients the finding was positive and in 25% the finding was suspect. The most frequent pathological change were microaneurysms; intraretinal microvascular abnormalities (IRMA) were detected in 13 patients. The trend towards proliferative changes is apparent in some patients surprisingly soon. The authors emphasize the sensitivity of the method which along with the extended classification of retinopathy makes its monitoring possible. The authors discuss the value of fluorescent angiography in the initial stages of retinal damage. They provide evidence of a statistically significant relationship of diabetic retinopathy and the duration of diabetes (p < 0.001) and age (p < 0.001).


[Immunologic reactivity in children on a chronic dialysis program and after kidney transplantation].

Dialysis and transplantation of the kidney have become in Czechoslovakia part of routine therapy of chronic renal failure (ChRF) in children and adolescents. The authors examined 16 children aged 4-17 years (x = 10.5) treated in a chronic dialyzation programme (ChDP) by haemodyalisis and 20 children after transplantation of a cadaverous kidney, age 4.5-16.5 years (x = 11.5), incl. 14 patients with a stable function of the graft and six patients during the period of acute rejection of the graft. The authors tested the basic parameters of humoral and cellular immunity in all three groups of children. In the group of the CRI a significant drop of the level was found only in serum IgA, the other parameters, incl. examination of sub-populations of lymphocytes CD3+, CD4+ and CD8+, did not differ significantly from normal values. In patients with a stabilized function of the graft treated by Cyclosporin A, prednisone, and in some cases also with azothioprim, the authors found a significantly lower absolute number of lymphocytes of the mentioned sub-populations as compared with controls and patients included in a ChDP. During the period of acute rejection the absolute number of CD3+, CD4+, CD8+ increased significantly, as compared with the period of stabilized function of the graft. Thus patients in CRI do not manifest despite the serious affection of the organism serious changes of immune reactivity within the scope of examinations made by the authors. Monitoring of immunity parameters in patients with transplants can contribute to evaluation of the effect of immunosuppression (it can draw attention to low dosage of drugs) and in the context with other clinical and laboratory examinations it can contribute to the diagnosis of acute rejection.


[High frequency "bubble" oscillation ventilation in the neonatal period].

The authors tested their own high frequency, oscillation, "bubble" ventilator on a model of neonatal lungs in vitro and in vivo. They used the effect of oscillations which develop when through fluid (in vivo water) a mixture of air is bubbled and which can be, if the expiration tube is sufficiently rigid, transmitted back into the lungs. If the CPAP circuit is suitably arranged, it can ensure an adequate exchange of blood gases. The authors found marked changes in the amplitude of pressure oscillations as well as of the frequency in relation to the shape of the ending beneath the fluid surface (greatest when a funnel with a 30 mm diameter and 90 degree angle in relation to a normal line projected on the water surface is used). They found that the compliance (C) has no substantial effect on the amplitude nor on the frequency of oscillations. The mentioned bubble system approached as to its efficiency the commercial oscillation ventilator Stephen 3000 SFH and was used in the therapy of respiratory failure in three neonates with body weights of 1250-2700 g. The authors conclude that high frequency oscillation "bubble" ventilation is a cheap, safe and highly effective therapy which can be used in any department providing care of neonates even during transport.


[Present possibilities of early diagnosis of hearing disorders in children belonging to the youngest age groups].

In a great proportion of children loss of hearing originates during the neonatal period. It is assumed that of 1000 neonates some 6 to 16 children suffer from impaired hearing. By early detection of hearing loss during neonatal and infant age by appropriate treatment, rehabilitation and training it is possible to achieve optimal speech development as well as development of mental and somatic abilities and to ensure to these children an adequate role in society. Early detection of impaired hearing based on screening of risk children already during the neonatal period is based on the use a risk registry. At the same time it is important to make parents and paediatricians of the first line of contact familiar with developmental stages of hearing and speech during the first year of life. Risk children must be dispensarized and they must be objected to orientative examination of hearing by reflex audiometry and other screening methods incl. the method of assessment of evoked otoacoustic emission. By systematic screening is is possible to detect suspected loss of hearing in as many as 60% of risk children. To verify the suspect impaired hearing it is necessary to make an objective examination of hearing by a child otolaryngologist.


[Variations in lipid metabolism in long-term monitoring of children treated for diabetes mellitus].

The authors investigated selected indicators of the lipid metabolism in 31 children with insulin-dependent diabetes for a period of 3 to 14 years. They divided the patients into two groups those with a glycosylated haemoglobin below or above 9 mumol/1 g haemoglobin. They compared the assembled results of the two groups and also with the results obtained in a control group. The total cholesterol was in both diabetic groups higher than in healthy subjects and was higher than the value which is considered from the aspect of the genesis and development of atherosclerosis as a risk value. Children with poorly compensated diabetes, i. e. with a glycosylated Hb level above 9 mumol had a higher cholesterol level as compared with well compensated diabetic children. The pre-beta fraction of lipoprotein increased in both groups of patients, however, more in those where the disease was not well compensated. There was a parallel decrease of the alpha fraction and the lipoprotein profile had a markedly atherogenic character. The apolipoprotein B concentration was in patients with well compensated diabetes lower, as compared with controls but in patients with poorly compensated diabetes after 4-5 years treatment is was significantly higher. Poor compensation of diabetes led after 4-5 years duration to a significant rise of the serum triglyceride level. As the blood lipid levels are influenced in a significantly way by diet, compensation of the disease and a low-fat diet are essential with regard to the results assembled in this investigation for prevention of ischaemic heart disease in diabetic subjects.(ABSTRACT TRUNCATED AT 250 WORDS)


[Growth, weight and physical proportionality in twins].

The growth, weight and proportionality of twins aged 1-15 years from the South Moravian Region were studied and compared with a control population. The sample of twins was divided in three groups: identical twins, nonidentical like-sexed twins and boys and girls from the unlike-sexed pairs. No reasonable differences between twins A and B were observed, therefore they are not evaluated separately. All the twins were born in years 1973-1987, the total number was 2,226 boys and 2,302 girls. A comparison with a control population revealed a lower mean height as well as weight of identical twins until the age of 15 years. The mean weight of identical twins was lower than that of nonidentical ones. Similar pattern were observed also in height, the statistical significance was found only in boys and only on a 5% level. Twins from unlike-sexed pairs were more like the control population than twins from unidentical like-sexed pairs, however, the differences in height in girls were not statistically significant. The distribution of proportionality values has shown an accumulation near zero--we can conclude that twins are more proportionate than other children of their age (i.e. less obese and less meagre). As expected, the observed intra-pair differences in identical pairs were significantly lower than in nonidentical like-sexed pairs.


[Pulsed magnetic fields--their possibilities in pediatric neurology].

The author reviews information on the use of a pulsatile magnetic field with defined parameters in some child diseases of the CNS according to his experience assembled during the past five years. PMP was applied in 17 cases with spinal amyotrophy type M. Werdnig-Hoffman and in 16 cases with DMO. In both diseases treatment was previously only symptomatic. PMP frequencies of alpha EEG waves were used during application on the area of the head and a different frequency for whole body treatment focused on muscular dystonia. The author draws attention to the specificity of biotropic parameters of the applied magnetic field.


[Occurrence of allergic diseases in children with neonatal allergic exanthema].

In 1990 in the allergological ambulatory department for children in Trencín 263 children were examined, born during the first half of 1981 and second half of 1982 who suffered during the first days after delivery from exanthema allergicum. In 186 children i.e. 71%, by the age of 8 and 9 years late allergies developed. The authors investigate various factors in the prenatal, perinatal and postnatal period and compare possible relations with the development of late allergic manifestation in children with exanthema allergicum neonatorum.


[Langerhans-cell histiocytosis in children].

Histiocytosis from Langerhans cells is a new term for a group of diseases formerly called histiocytosis X. In The Faculty Hospital Motol the authors treated between July 1974 and December 1980 84 children with this disease. Twenty one suffered from the malignant form (formerly Letterer-Siwe and Hand-Schüller-Christian disease) and in 63 patients the diagnosis of eosinophil granuloma was established. In 21 children with unequivocally malignant disease the authors started chemotherapy immediately after establishment of the diagnosis and in six they indicated in addition radiotherapy. Of these 21 children 16 are in complete remission and 5 children died from progression of the disease during treatment. In the group of 63 children with eosinophil granuloma in 44 only excochleation of the focus was performed. Chemotherapy was administered to 12 children, incl. 5 where it was combined with radiotherapy. A relapse of the disease was recorded in 8 children. At present all patients suffering from the disease are in complete remission.


[Transitory hyperphosphatasemia in childhood].

Transient hyperphosphatasaemia was detected in 11 children hospitalized at the First and Second Paediatric Clinic of the Paediatric Faculty Hospital in Bratislava in 1985-1990. The authors analyzed retrospectively 6 children from this group where it was possible to evaluate accurately the trend of serum alkaline phosphatase. In all children the high alkaline phosphatase activity was detected incidentally during the initial examination. The reason for hospitalization were in four instances respiratory infections, in one instance coeliac disease and in one instance infectious mononucleosis with infection of the urinary pathways. The maximum increase of serum alkaline phosphatase was 5-15 fold higher than the upper borderline of the reference range for the given age group. The activity declined to normal spontaneously independently on the course of the basic disease and treatment, always in the course of 3-12 weeks. The isoenzyme pattern of alkaline phosphatase with a high ratio of the thermolabile isoenzyme was typical. Finally the authors emphasize that recognition of this obscure condition which does not endanger life can spare the children many unnecessary and expensive examinations.


[Liver diseases in children with alpha 1-antitrypsin deficiency in infancy].

The authors evaluate the health status of children with alpha-1-antitrypsin deficiency, focused on liver disease in infant age. The children were selected by neonatal screening. Of 21 children one had severe neonatal hepatitis with progression to cirrhosis, 2 children had clinically apparent jaundice to the age of two months, 6 children had elevated total bilirubin and transaminase levels without clinical signs of the disease, 12 of the remaining children had no clinical and laboratory signs of liver disease. In the discussion the authors compare the results with data published abroad.


[Carbocysteine in the treatment of recurrent bronchitis in infants].

In a group of 51 children aged 6-24 months the therapeutic effectiveness of the mucolytic preparation carbocysteine was tested and compared with the effect of Ipeca syrup. The effect was evaluated by means of a point score comprising changes of the clinical picture of the disease and the use of other laboratory examinations. The results of the examination revealed the more favourable effect of carbocysteine, as compared with a mixture of Ipeca syrup in the treatment of acute relapsing bronchitis in infants.


A case of hyperplasia of the anterior lobe of the pituitary imitating intrasellar expansion.

The authors describe the case of an obese girl with severe hypothyroidism which led to increased TSH secretion and finally to hyperplasia of the anterior lobe of the pituitary diagnosed by magnetic resonance. After substitution treatment with thyroid hormones was started, marked improvement of the general condition, loss of body weight and normalization of laboratory findings and regression of hyperplastic tissue occurred.


[Sixth disease--exanthema subitum].

The author deals with the epidemiology of the sixth disease--exanthema subitum which is caused by the herpes virus hominis type 6. There are nine forms of the disease. The haemogram is characterized by granulocytopenia, eosinopenia with relative lymphocytosis, thrombocytopenia.


[Multiple (serial) poisoning with scopolamine present in a compounded nose drop preparation].

The authors describe three cases of scopolamine intoxication which was added to nasal drops instead of Mucoseptonex. An account is given of the characteristics of scopolamine intoxication and of possible therapy.


[The capacity of IL-2 production of peripheral blood lymphocyte in patients with pneumonia].

The capacity of IL-2 production of peripheral blood lymphocyte (PBL) were determined in 54 patients with pneumonia in acute stage (23 cases of 20-57 years old and 31 cases over 60 years old) and 33 cases in convalescent stage. 20 elderly COPD in remission patients and 59 healthy control (32 aged 20-55 and 27 over 60) were determined also. It was shown that the capacity of IL-2 production of the patients with pneumonia in acute stage were markedly lower than healthy control (P less than 0.01), and that of the elderly patients' were markedly lower than elderly COPD patients also (P less than 0.01). The IL-2 level were also lower in the patients with pneumonia in convalescence than in the healthy control (P less than 0.01). The capacity of IL-2 production of elderly COPD patients and healthy elderly donors were at the same level (P greater than 0.05), both of the two groups' IL-2 level were significantly lower than that of the healthy young-middle age donors. It was suggested that the lowered immune function may be one of the important factors causing the COPD patients and the elderly people susceptible to infection, and it may be one of the important causes of the more severe condition and protracted course in elderly patients.


[A primary study on the relationship between the acute infection of chronic obstructive pulmonary disease (COPD) and mycoplasma pneumoniae].

The specific antibodies of Mycoplasma Pneumoniae (MP) in the sera of 227 normal adult controls and 187 adult patients with acute infections of COPD were measured by immunofluorescence assay (IFA). The findings showed: 1. in the sera of control group, the titer of specific IgM antibody of MP were lower than 1:8 in 224 cases (98%) and the specific IgG antibody were below 1:16 in 218 cases (96%). 2. Positive results for MP antibodies (the IgM 1:8 or/and the IgG 1:16) were found in 70 cases (37.4%) of adult patients with acute infections of COPD. We suggest that Mycoplasma Pneumoniae is a common pathogen for the adult acute infections of COPD.


[Pulmonary Aspergillus infection. A study of 61 cases of clinical, roentgenologic and pathological findings].

The clinical results of 61 patients with pulmonary aspergillosis were analysed. Patients were admitted since 1975 to 1989. Pathologic evidences were obtained at postoperation in 51 patients. The main clinical symptom was hemoptysis, and was frequently misdiagnosed as pulmonary tuberculosis, bronchiectasis or other pulmonary disease. The diagnosis and treatment of pulmonary aspergillus infection was discussed.


[Pathological changes in the trachea after tracheostomy in patients with respiratory failure].

A variety of pathological changes in trachea were observed in 7 cases with tracheostomy. The common lesions were inflammation, necrosis and ulcer, and squamous metaplasia or granuloma were seen in those with tracheal tube for longer time. The dilatation of trachea was manifested by dislocating of the tube and not sealing of the cuff. Cuff-radiography could show the site and degree of the dilatation of trachea. In order to alleviate the lesions, the tubes made of plastic or silica gel with lower pressure cuff were recommended. The prevention of local infection and improvement of nutritional status were also important.


[A study on the functional condition of airway muscarinic M2-receptors in asthma].

We investigated the hypothesis that airway M2-receptor selectively pretreated with aerosolized pilocarpine would modify the bronchoconstrictor response to histamine, which is, in part, vagally mediated. On two different days, the following two histamine inhalation challenges were performed in 12 normal individuals and 13 stable asthmatics: histamine alone or pilocarpine-histamine. In normal subjects, prior M2-receptor stimulation with pilocarpine suppressed the subsequent bronchial response to histamine. In asthmatic patients, however, prior pilocarpine exposure failed to modify the bronchial response to histamine. The results suggest that prior M2-receptor stimulation has a protective effect on histamine induced bronchoconstriction in normal subjects and the absence of this inhibitory effect in asthmatic patients may represent the existence of functional depression of M2-receptors in asthmatic airways.


[The effect of grain dust on non-specific bronchial hyperreactivity].

246 grain elevator or mill workers were divided into heavy-exposure group and light-exposure group and methacholine provocation tests, were carried out. After provocation test, FEV1, FVC, MMEF, PEF, V50 and V25 significantly decreased in heavy exposure than those in light exposures. (P less than 0.01). 18.9% of heavy-exposure was associated with bronchial hyperreactivity, which was significantly higher than that of light-exposure (4.4%). The incidence of more than 10% declines in FVC or FEV1 and 25% in PEF or V25 during one day or one week work-shift was significantly higher in workers with bronchial hyperreactivity than in those without bronchial hyperreactivity (P less than 0.01). The findings indicated that exposure to heavy concentration of grain dust for long period can impair pulmonary function and cause nonspecific bronchial hyperreactivity.


[The treatment of bronchiectasis by using left lower lobectomy with extirpation of lingular division of the bronchi].

7 cases with bronchiectasis of left lower lobe and lingular segment were treated with left lower lobectomy and extirpation of the bronchi of lingular segment. Its advantages are that the upper division lung will not be damages, the complication of sigmentectomy be avoided. Meanwhile, the ability of lung expand will be remained and the compensating emphysema were prevented in the survival lung. In addition, the operating indication and method were discussed.


[Studies on whole chromosomal DNA probes from Mycobacterium tuberculosis and Mycobacterium intracellulare].

The whole chromosomal DNA probes prepared from M. tuberculosis and M. intracellulare were used to hybridize in DNA dot blot and bacterial dot blot hybridization with 9 reference strains of Mycobacteria, including M. tuberculosis, M. bovis, M. avium, M. intracellulare, M kansasii, etc. These two probes showed good sensitivity and specificity for M. tuberculosis complex and M. avium complex respectively. Using the colony hybridization in situ, we can detect and identify M. tuberculosis with the probes on NC membrane after 1-2 week incubation over 7H9 agar plate.


[The study of interleukin-2 and interleukin-2 receptor in patients with pulmonary tuberculosis].

IL-2 level and IL-2R expression of peripheral blood lymphocytes (PBL) were measured in 25 patients with type III and advanced tuberculosis (TB). The results showed that IL-2 level in patients was lower than that in controls, and that in the patients whose history of TB was over 3 years was significantly lower than that in the patients whose history of TB was within 3 years; There was no significant difference between the IL-2R expression on unstimulated PBL from patients and controls. IL-2R expression in PHA-stimulated PBL after culturing 72 hours in patients was significantly lower than that in controls, and that in the patients over 3 years was significantly lower than that in the patients within 3 years. The data indicated that the function of T helper cells in patients with TB was impaired, and the impairment was more serious in the patients with longer illness.


[The diagnosis of smear negative pulmonary tuberculosis in superior segment of lower lobe].

20 cases of tuberculosis in the superior segment of the lower lobe of the lung were misdiagnosed as lung cancer, pneumonia, bronchiectasis and inflammatory pseudoneoplasm were reported. The final diagnosis were confirmed by fiberoptic bronchoscopy (FOB). The causes of the misdiagnoses were: (1) the hilar mass shadow found on the PA chest film, mistaken for central type lung cancer; (2) the mass shadow found on the lateral chest film, mistaken for peripheral lung cancer; (3) the patients with fever, cough and expectoration accompanied by exudative infiltrative shadow, mistaken for pneumonia; (4) patients with recurrent attacks of hemoptysis but the lesions overshadowed by the spinal column on the lateral chest film were misdiagnosed as bronchiectasis. The author suggested PA and lateral chest films taken simultaneously were needed. The special points, to which should be pay attention during reading the films were listed and noted. FOB examination including TBLB, brushing and BALF for pathologic and AFB determination could be of help to confirm the diagnosis.


[Clinical analysis of chronic renal function failure accompanied by pulmonary tuberculosis].

In 592 patients with chronic renal function failure (CRFF), 31 were accompanied by pulmonary tuberculosis, the incidence was 5.2%. The incidence of the First Affiliated Hospital of Sun Yat-sen University (6.6%) is higher than that of Zun Yi District Hospital (3.9%). The difference may be associated with dialysis therapy and regional TB natural population incidence. The data also showed that accompanied TB may be a reversible factor of deteriorating CRFF. Details of diagnosis and treatment of CRFF accompanied by TB are presented.


[Rapid diagnosis of pulmonary Pseudomonas aeruginosa infections by indirect immunofluorescent antibody].

167 sputum specimens collected from patients with pulmonary infection were tested by IFA (Indirect immunofluorescent antibody-staining) with serogroup-specific monoclonal antibody to Pseudomonas aeruginosa, compared with quantitative sputum culture. The results showed the minimum concentration of bacteria detectable by IFA was 10 cfu/ml. 31(86.1%) were positive in 36 specimens with more than 10 cfu/ml of Pseudomonas aeruginosa. 22(100%) were positive in 22 specimens with more than 10 cfu/ml of Pseudomonas aeruginosa. In 127 specimens with a negative culture for Pseudomonas aeruginosa, 121(95.2%) were negative, 6(4.8%) were false-positive which could be identified by Gram's-staining. The type of Pseudomonas aeruginosa could also be identified within 3 hours.


[CT studies on obstructive sleep apnea syndrome].

To ascertain whether there are structural and functional changes in the upper airway of obstructive sleep apnea syndrome (OSAS) pts. We studied 33 pts of OSAS with positive polysomnographic tests, 14 pts of non-apnea snore with negative polysomnographic tests, and 18 normal subject by using CT scanner (Somatom Dr 3). The results are as followings 1. There were narrowing region in the upper airway in OSAS pts and the non-apnea snore pts. Most of the narrowings were located at the level of oropharynx, which was correspondent to the level of the soft palate and the uvula. The length of the soft palate and the pre-spinal soft tissue thickness of two groups of pts were larger than non-snore individual. 2. The pharyngeal compliance was significantly higher in OSAS pts than in non-apnea snore pts. 3. There was no localised deposition of fat tissue surrounding the lumen of upper airway. 4. CT scanning is the non-invasive and more useful procedure for the diagnosis of OSAS and also would help to select the pts for surgery.


[Effect of enoxacin on theophylline pharmacokinetics].

The effect of a multiple-dose regimen of oral Enoxacin on Theophylline pharmacokinetics was evaluated in 10 hospitalized patients with COPD. It was found that mean Theophylline concentrations in serum significantly increased 144% (P < 0.01), AUC increased 350.4% (P < 0.01), T1/2K increased 204% (P < 0.01), clearance decreased 76.8% (P < 0.01) during coadministration of enoxacin than before. This results suggested that a multidose regimen of enoxacin significantly slowed the clearance of theophylline and elevated theophylline concentrations in serum. The careful monitoring of serum theophylline level and modification of theophylline dosage in patients receiving enoxacin and theophylline were recommended.


[The third nationwide random survey for the epidemiology of tuberculosis in 1990].

The third nationwide epidemiological survey for tuberculosis covered 29 provinces, municipalities and autonomous states in the Country. A total of 928 survey points were surveyed. The results of the survey showed the following facts: The prevalence of pulmonary tuberculosis (radiological active) was 523/100,000 population; the smear-positive prevalence was 134/100,000; The mortality of tuberculosis was 21/100,000; The tuberculous death ranked the seventh among ten leading causes of death. The results of the survey also showed that the decline of the magnitude of tuberculosis was so slow. The decline of the magnitude of tuberculosis in the coast provinces were much faster than the inland and remote provinces and it was faster in cities than in rural areas.


[Application of enzyme linked immuno-electrophoresis for the diagnosis of tuberculosis].

The technique of Enzyme linked immuno-electrophoresis (ELIEP) was established to detect the specific antibody against tubercle bacilli. The results of 730 cases showed that the sensitivity of this method is 97.33%, and the specificity, 97.5%. Thus this method is more sensitive than the ELISA methods which had bean published. The manipulation of ELIEP is simple, convenient, and fast, reagent stable. This method may be used in diagnosis of tuberculosis with reliability and efficiency.


[An analysis of lymphocyte subsets and purified protein derivative-antibody in pulmonary tuberculosis].

The lymphocyte subsets and PPD-antibody in peripheral blood were determined in 72 cases of bacteriological positive pulmonary tuberculosis using monoclonal antibody to anti-T lymphocyte and ELISA assay. The results showed that the percentage of T3 was 52.44 +/- 11.46% and the OD of PPD-antibody was 0.47 +/- 0.27 in initially treated patient group; 40.27 +/- 7.81%, 0.81 +/- 0.17 in chronic carrier group respectively. There were statistically significant difference between the two groups (P < 0.01). The percentage of T3 was significantly larger in 30-59 yr group than in more than 60 yr group among non chronic carriers (55.9 +/- 11.3% vs 44.4 +/- 2.4, P < 0.05). PPD-antibody was strongly associated with extent of lesion in lung.


[Arteriography and bronchial artery embolization for massive hemoptysis in 100 cases of tuberculous].

Bronchial arteriography and embolization with sponge gelatin were performed in 100 tuberculous patients of massive hemoptysis. Three direct signs and other six indirect ones were found in arteriographies, Those signs were roentgenographic abnormalities for defining bleeding sites. 100 massive hemoptysis cases were treated with sponge gelatin embolization in bronchial artery. The results are satisfactory, the success rate were 92.9%, 87.4% and 79.0% at one week, one month and three months respectively. The advantages of arterial embolization and the reasons of failures were discussed in this paper.


[Evaluation of the treatment of adult tuberculous meningitis].

Through analysis of the treatment in 205 patients of adult tuberculous meningitis, we considered: 1. effective and vigorous alleviation of high intracranial pressure in early stage, 2. appropriate dosage of corticosteroids and, 3. correct selection of antituberculous drugs was the mainstay for chemotherapy of tuberculous meningitis. The results were 82 patients (40.0%) were cured, 101 cases (49.3%) improved, 15 cases (7.3%) died and 7 cases (3.4%) had complications. 175 cases have been followed-up from two to five years, all of them are alive.


[Experience of focal debridement in the treatment of tuberculosis of lumbar spine].

Experiences of 882 cases with tuberculosis of lumbar spine treated by focal debridement during 1956-1986 all patients survived the operation and 661 patients were cured in our department were reported. The relationship between chemotherapy and focal debridement was discussed. The selection of the appropriate time for this operation and important points to which attention should be paid during operation were described in detail.


[The pathological changes in pulmonary vessels complicated by cirrhosis with anatomicopathological analysis of 30 cases].

The pulmonary sections HE stained in 30 autopsy cases of cirrhosis were randomly reexamined. The distribution and degeneration of pulmonary arterioles and venules which were at alveolar level and the diameter of capillaries were observed; 10 autopsy cases of other diseases were examined as control. The results showed tricuspid orifices were dilated in 20% of autopsies; Both lungs were heavier than those in the control group, pulmonary congestion were in 100% of autopsied lungs. Pulmonary artery degeneration occurred in 16.6%, the number of pulmonary arterioles increased in 73.0%, the hypertrophic dilatation and increasing number of pulmonary arterioles in 33.3%; local spider in pulmonary pleura in 20%. We conclude that above findings will be helpful to the indirect diagnosis of intra-pulmonary shunt and pulmonary hypertension.


[Changes and significance of chemiluminescence of polymorphonuclear leukocytes in endotoxin-induced lung injury in conscious sheep].

The chemiluminescence (CL) of polymorphonuclear leukocytes and its relation with pulmonary microvascular permeability after endotoxin-induced lung injury in conscious sheep with lung lymph fistula were observed. Four hours after the injury the CL of PMNs increased from 0.27 cpm/PMN of baseline to 0.69 cpm/PMN (P < 0.05). The increment of the CL had positive correlation with the increment of lung lymph flow or permeability index (r = 0.632 0.638 P < 0.05), suggesting that the increase of pulmonary microvascular permeability after the endotoxin injury had relation with the increase of the respiratory tract of PMNs.


[Changes in activity of phospholipase A2 and its metabolic production of prostaglandins in lung tissue of asthmatic guinea pigs].

The activity of phospholipase A2 and its metabolic production-prostaglandins were examined in lung tissue of allergic asthmatic guinea pigs. The results shows that the activity of phospholipase A2 after three days of asthma attack was raised 54.19% than that of control group (P < 0.01), and decreased 69.27% when protected with chloroquine before asthma, and decreased 65.98% when only given chloroquine as a control; the ratio of those prostaglandins was resumed to control group when previously given chloroquine in asthma and normal guinea pigs. These results suggest that the activity of phospholipase A2 and changes of prostaglandins in lung tissue were related to allergic asthma of guinea pigs, the increase of TXB2 or PGF2 alpha and decrease of PGE2 or 6-Keto-PGF2 alpha in lung tissue after three days of asthma attack may have a correlation with allergic asthma in guinea pigs.


[Qualitative and quantitative observations of smoking-induced morphologic changes in muscular pulmonary arteries].

Light microscopy, transmission electron microscopy, image processing and morphometric technique were applied to analyze the morphologic changes in the muscular pulmonary arteries from the rats after exposure to smoking. The results showed that the pronounced muscularization of the small pulmonary arteries. significant reduction of the intraacinar arteries, intimal and medical thickening that was significantly associated with the cellular elements and extracellular matrix, and the smooth muscle cells proliferated and migrated from the media to intima of artery, the cell shape modified and increased in rough endoplasmic reticulum also were found. Although the functional significance of these findings is unknown, they might play an important role in smoke-induced occurrence and development of pulmonary hypertension and cor pulmonale.


[Short-term effect and the mechanism of radix Angelicae on pulmonary hypertension in chronic obstructive pulmonary disease].

The changes of hemodynamics, hemorheology and blood gas were investigated in 28 stable COPD patients with pulmonary hypertension after short-term treatment by Radix Angelicae. The level of TXA2, PGI2 were measured in 10 of the 28 patients before and after treatment. The results show that Radix Angelicae produced significant improvement on pulmonary hypertension, blood viscosity, hematocrit and the level of TXA2 decreased significantly. But the blood gas, blood pressure and the level of PGI2 remained unchanged in all of the patients.


[Changes in acetylcholine and acetylcholinesterase in blood during pulmonary hypertension].

Two models of pulmonary hypertension in rats have been made with either monocrotaline or passive smoking (with or without infection). The decrease of RBC acetylcholinesterase activity and the increase of acetylcholine content in blood were detected in both rat models, but did not in rats which were only once infected by E. Coli, and the pulmonary pressure of the latter did not raise either. Comparing 30 cases of acute attack period of chronic pulmonary heart disease with 30 cases of normal persons, RBC acetylcholinesterase decreased significantly, indicating that hyperfunction of cholinergic nerve existed in pulmonary hypertension and pulmonary heart disease.


Influence of circadian light-dark alternations on macrophages and lymphocytes of CBA mouse.

The influence of circadian 12 h light-12 h dark alternations on CBA mouse macrophages and lymphocytes was determined using tests for macrophage spreading and ingestion ability or flow cytometry immunophenotyping of blood, lymph node, and spleen lymphocytes. The animals were tested every 4 h around the clock. Collected macrophages were incubated in vitro for 3 or 18 h. Monoclonal antibodies permitted detection of T-lymphocytes, suppressor-cytotoxic T-lymphocytes, helper-inducer T-lymphocytes, or B-lymphocytes. Two types of analyses were performed: First, the difference between the same intervals of the 12 h light or dark period was determined. The macrophage ingestion was significantly lower at the beginning and higher at the end of the dark period. We have also found a significant increase in blood T-lymphocytes of helper-inducer T-lymphocyte percentages and of the T helper-inducer: T suppressor-cytotoxic ratio during the dark period. Second, the ultradian variation during the 12 h light or dark period was determined. The variability was significant both for macrophage spreading and ingestion. Multiple significant variations of lymph node, spleen, or blood lymphocyte percentages were also observed. All of these data indicate that daily alteration of the lighting regimen significantly influences mouse peritoneal macrophage functions and various lymphocyte subsets.


Daily rhythms of metabolic rate and body temperature of two murids from extremely different habitats.

Daily circadian rhythms of body temperature (Tb) and oxygen consumption (VO2) were measured in two murid species, which occupy extremely different habitats in Israel. The golden spiny mouse (Acomys russatus) is a diurnal murid distributed in arid and hot parts of the great Syrio-African Rift Valley, while the broad-toothed field mouse (Apodemus mystacinus) is a nocturnal species that inhabits the Mediterranean woodlands. In both species, the daily rhythms of Tb and VO2 are entrained by the photoperiod. Under laboratory experimental conditions (ambient temperature Ta = 33 degrees C and photoperiod regime of 12L:12D), Acomys russatus exhibits a tendency towards a nocturnal activity pattern, compared to the diurnal activity displayed by this species under natural conditions. Under the same photoperiod regime and at Ta = 28 degrees C, Apodemus mystacinus displays nocturnal activity, as observed under natural conditions. The maximal values of Tb were recorded in Acomys russatus at midnight (23:50 h), while the maximal values of VO2 were recorded at the beginning of the dark period (18:20 h). In Apodemus mystacinus, the maximal values of Tb and VO2 were recorded at 23:40 and 20:00 h, respectively. The ecophysiological significance of these results is discussed further.


The in vitro effect of metyrapone on steroid synthesis in mice adrenals at different circadian stages.

The circadian rhythm of the in vitro biosynthesis of cortisol and cortisone in mice adrenals has been documented in the absence and presence of 0.1 mumol metyrapone, an inhibitor of steroid 11 beta-monooxygenase. After 3 weeks of synchronization with 12 h light:12 h darkness, adrenalectomy was performed at eight circadian stages: 0, 4, 9, 10, 13, 16, 21, and 22 h after light onset (HALO). Because it has been shown that mice adrenals could convert exogenous 11-deoxycortisol, the synthesis of 11-oxysteroids (cortisol+cortisone) in adrenal homogenates was studied from tritiated precursor. The pattern of steroid synthesis showed a maximum around the end (10 HALO) and a minimum at the beginning of the resting period (0 HALO); the variation was approximately 10%. A similar pattern was observed in the presence of a approximately 50% inhibiting dose of metyrapone. On the other hand, the percent inhibition of 11-oxysteroids synthesis was greater at the beginning of the resting period (0 HALO) and minimum around the end of the activity span (21 HALO), with an overall variation of 20%. However, the variations were statistically insignificant (unpaired t test).


Vasopressin antagonist disrupts the circadian rhythm of water intake on suprachiasmatic injection.

The present study makes an attempt to find out the action of arginine vasopressin (AVP) and its antagonist d-(CH2)5 Tyr (Me) AVP applied at the suprachiasmatic nuclei (SCN) on the circadian rhythm of water intake. Chronic implantation of a 22 G stainless steel cannula for injection was performed using a stereotaxic technique under Nembutal anesthesia. AVP and its antagonist were injected into the SCN of free-moving rats at the beginning of light and dark phases of the light-dark (LD) cycle. Injections of AVP during either phase did not disrupt the circadian pattern of water intake while the injections of the antagonist disrupted it. The findings are suggestive of the involvement of AVP as a mediator of the circadian rhythm of water intake at the level of the neural pacemaker, SCN.


The variability in circadian phase and amplitude estimates derived from sequential constant routines.

Both the constant routine (CR) and the dim light melatonin onset have been suggested as reliable methods to determine circadian phase from a single circadian cycle. However, both techniques lack published studies quantifying the intercycle variability in their phase resolution. To address this question eight healthy male subjects participated in two CRs, 7 days apart. Circadian phase was determined using 3-min samples of core body temperature and two hourly urinary sulphatoxy melatonin excretion rates. Phase and amplitude were estimated using simple (24 h) and complex (24 + 12 h) cosinor models of temperature data and the onset, offset, and a distance-weighted-least-squares (DWLS) fitted acrophase for the melatonin metabolite. The variability in phase estimates was measured using the mean absolute difference between successive CRs. Using the simple 24 h model of temperature data, the mean absolute phase difference was 51 min (SD = 35 min). Using the complex model, the mean absolute phase difference was 62 min (SD = 35 min). Using the DWLS fitted acrophase for the melatonin metabolite, the mean absolute phase difference between CR1 and CR2 was 40 min (SD = 26 min). The results indicate that for CRs a week apart, the mean absolute difference in an individual's phase estimate can vary by 40-60 min depending on the choice of dependent measure and analytic technique. In contrast to the intraindividual variability, the group results showed considerably less variability. The mean algebraic difference between CRs, using temperature- or melatonin-derived estimates, was less than 5 min, and well within the range of normal measurement error.


Rhythmic and nonrhythmic modes of anterior pituitary gland secretion.

Because of confounding effects of subject-specific and hormone-specific metabolic clearance, the nature of anterior pituitary secretory events in vivo is difficult to ascertain. We review an approach to this problem, in which deconvolution analysis is used to dissect the underlying secretory behavior of an endocrine gland quantitatively from available serial plasma hormone concentration measurements assuming one- or two-compartment elimination kinetics. This analytical tool allows one to ask the following physiological questions: (a) does the anterior pituitary gland secrete exclusively in randomly dispersed bursts, and/or does a tonic (constitutive) mode of interburst hormone secretion exist? and (b) what secretory mechanisms generate the circadian or nyctohemeral rhythms in blood concentrations of pituitary hormones?(ABSTRACT TRUNCATED AT 250 WORDS)


Melatonin shifts human circadian rhythms according to a phase-response curve.

A physiological dose of orally administered melatonin shifts circadian rhythms in humans according to a phase-response curve (PRC) that is nearly opposite in phase with the PRCs for light exposure: melatonin delays circadian rhythms when administered in the morning and advances them when administered in the afternoon or early evening. The human melatonin PRC provides critical information for using melatonin to treat circadian phase sleep and mood disorders, as well as maladaptation to shift work and transmeridional air travel. The human melatonin PRC also provides the strongest evidence to date for a function of endogenous melatonin and its suppression by light in augmenting entrainment of circadian rhythms by the light-dark cycle.


The cellular and molecular basis for divergent allergic responses to chemicals.

Chemicals vary with respect to the nature of allergic reactions which they will elicit preferentially. A wide variety of environmental and industrial chemicals are known to cause allergic contact dermatitis (contact sensitivity). Some of these are able also to induce respiratory allergy. This article reviews the characteristics of immune responses to different classes of chemical allergens and the role which functional subpopulations of T helper (TH) cells and their soluble cytokine products play in the induction of allergic sensitization. In addition, new opportunities to identify and classify chemical allergens based upon characterization of divergent allergic responses is discussed.


32P-postlabeling of acrolein-deoxyguanosine adducts in DNA after nuclease P1 digestion.

In order to study the relationship between the level of acrolein-DNA adducts and their biological effects, sensitive methods are needed to quantitate DNA adducts. 32P-postlabeling is one such method that has been widely used and we have adapted the technique to detect acrolein-deoxyguanosine adducts. Adducts formed by the reaction of acrolein and deoxyguanosine-3'-monophosphate were isolated by HPLC. Based on their UV spectra and cochromatography with standards after dephosphorylation with acid phosphatase, these adducts were identified as the nucleotide equivalents of cyclic 1,N2-propanodeoxyguanosine adducts formed by acrolein that have been described by Chung et al. [15]. As nucleotides, the adducts were good substrates for polynucleotide kinase-mediated transfer of phosphate from ATP and were able to be detected by 32P-postlabeling. These adducts were resistant to the activity of nuclease P1 and dinucleoside monophosphates in the form d(G*pN) where G* is the acrolein-guanine adduct also resisted digestion by nuclease P1. Digestion of DNA by nuclease P1 and acid phosphatase resulted in the conversion of normal nucleotides to nucleosides and selective enrichment of the adducts as dinucleoside monophosphates. Using nuclease P1/acid phosphatase digestion, followed by 32P-postlabeling and TLC separation, levels of the two adducts in acrolein-treated DNA were found to be about 6185 and 19,222 nmol/mol.


Effect of fluorine substitution on benzo[j]fluoranthene genotoxicity.

The metabolism and mutagenic activity of 4-fluorobenzo[j]fluoranthene (4F-B[j]F) and 10-fluorobenzo[j]fluoranthene (10F-B[j]F) were evaluated and compared with benzo[j]fluoranthene (B[j]F) using an identical rat liver homogenate preparation. Previous studies have shown that the major genotoxic metabolites of B[j]F are the 4,5- and 9,10-dihydrodiol. The 9,10-dihydrodiol was the principal metabolite formed in the case of 4F-B[j]F, while the 4,5-dihydrodiol was the principal metabolite formed in the metabolism of 10F-B[j]F. Studies on the relative genotoxicity of these fluorinated derivatives were performed to indirectly determine the possible contribution of the 4,5- and 9,10-dihydrodiol in the activation of B[j]F to a genotoxic agent. In the presence of microsomal activation, both of these fluorinated derivatives of B[j]F were more mutagenic in S. typhimurium TA97a, TA98 and TA100 than B[j]F. However, differences in mutagenic potency were observed between 4F- and 10F-B[j]F. 10F-B[j]F had similar mutagenic potency to 4F-B[j]F in TA97a and TA98 at doses associated with the linear portion of the dose response curve. However, a slightly higher mutagenic response was observed with 10F-B[j]F in TA98 at doses above 5 nmol. In contrast, 4F-B[j]F was more active than 10F-B[j]F as a mutagen in TA100. The tumor-initiating activity of these analogs on mouse skin was assessed at doses of 2.0, 1.0 and 0.3 mumol. Skin irritation was observed with the fluorinated B[j]F derivatives at doses above 0.3 mumol. At a dose of 0.3 mumol, 4F-B[j]F exhibited tumorigenic activity which was similar to B[j]F. In contrast, 10F-B[j]F was less active than B[j]F at all three doses assayed. Both fluorinated derivatives of B[j]F formed higher levels of DNA adducts in vivo in mouse skin than B[j]F. A modified 32P-postlabeling method was required to detect fast migrating B[j]F:DNA adducts that went undetected in previous studies. The level of DNA adducts formed from 4F-B[j]F was considerably greater than the levels observed with 10F-B[j]F. This is consistent with the greater mutagenic activity in S. typhimurium TA100 and tumor-initiating activity exhibited by 4F-B[j]F. These studies suggest that fluorine substitution may significantly alter the intrinsic genotoxicity of the 4,5- and 9,10-dihydrodiol of B[j]F. These data also imply that B[j]F may be primarily activated via the formation of the 9,10-dihydrodiol metabolite. This pathway of activation is inconsistent with our previous studies which indicate that the 4,5-dihydrodiol is the most important pathway of activation.(ABSTRACT TRUNCATED AT 400 WORDS)


Characterisation and quantitation of a selenol intermediate in the reaction of ebselen with thiols.

The reaction of ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) with thiols was investigated with particular attention to the formation of an ebselen selenol intermediate. The selenol intermediate could be trapped in a mixture of ebselen and thiols with 1-chloro-2,4-dinitrobenzene and the resulting product displayed unique spectral characteristics. The reaction of authentic, synthesised ebselen selenol with 1-chloro-2,4-dinitrobenzene (CDNB) was shown to give rise to the same compound (2,4-dinitrophenyl (N-phenyl-2-carboxamido phenyl) selenide as characterized by light spectroscopy, NMR, IR and elemental analysis. The determination of the absorbtion coefficient at 400 nm (E = 7.5 mM-1 cm-1) and the initial rate constant of the reaction (1.4 +/- 0.3 mM-1 min-1) allows for the convenient quantification of ebselen selenol concentrations by initial rate measurements after addition of CDNB. The choice of 400 nm to monitor the reaction excludes the interference of other intermediates in the reaction of ebselen with thiols as well as the reaction of the thiols with CDNB. When the assay is applied to typical incubation conditions used for investigating the glutathione peroxidase-like activity of ebselen it was shown that as much as 10-20% of ebselen is in the selenol form. If a stronger reductant (dithiothreitol) is used 60% is in the selenol form. These data could also be confirmed by the direct determination of ebselen selenol by UV spectroscopy, due to its peak absorption at 370 nm (E = 2 mM-1 cm-1). In conclusion, this investigation demonstrates, for the first time, the identity and quantity of ebselen selenol in the reaction of ebselen with thiols and also describes a convenient assay for its quantification. These observations allow further possibilities for investigation of the molecular species responsible for the antioxidant and peroxidase activities of ebselen.


Determination of the relative contributions of the diselenide and selenol forms of ebselen in the mechanism of its glutathione peroxidase-like activity.

The molecular basis of the glutathione peroxidase activity of ebselen (2-phenyl-1,2-benzisoselenazol-3(2H)-one) was investigated by the use of synthesised, authentic intermediates identical to those formed by the reaction of ebselen with glutathione. The second order rate constants for the reaction of ebselen (0.29 mM-1 min-1), ebselen-glutathione selenosulfide (less than or equal to 0.01 mM-1 min-1), ebselen selenol (2.8 mM-1 min-1) and ebselen diselenide (0.32 mM-1 min-1) with hydrogen peroxide reveal that the selenol is particularly active in this respect. The determination of the relative amounts of ebselen selenol and diselenide under typical peroxidase assay conditions implies that the selenol is the predominant molecular species responsible for the glutathione--(70%)--and dithiothreitol--(96%)--dependent peroxidase activity of ebselen.


The effect of six sesquiterpenoid unsaturated dialdehydes on cell membrane permeability in human neuroblastoma SH-SY5Y cells.

The effect of six sesquiterpenes containing an unsaturated dialdehyde functionality, on cell membrane permeability in the human neuroblastoma cell line SH-SY5Y has been studied. The kinetics of the membrane leakage after addition of the sesquiterpenes were determined by measuring the efflux of radioactivity from cells preloaded with tritiated 2-deoxyglucose. The concentrations that gave 5% and 20% efflux of radioactivity as compared with control cells (EC5 and EC20) were determined for each compound. In spite of the structural similarities between the compounds, the effects on cell membrane permeability varied considerably. EC20 for polygodial, which is the most active compound, is 2.5 microM after 20-min incubation, but no leakage could be determined for merulidial even at concentrations as high as 4 mM. Rather, this compound seems to stabilize or fix the cell membrane and a lower efflux of radioactivity was observed as compared to the control cells. A quantitative structure-activity relationship analysis for the five active compounds showed a good correlation between the membrane leakage activity and certain chemical characteristics. Structural features strongly correlated with high activity were found to be: The geometry and the atomic charges of the unsaturated dialdehyde functionality, the dipole moment, the energy difference between the lowest unoccupied molecular orbital and the highest occupied molecular orbital and the lipophilicity.


The relative importance of glutathione and metallothionein on protection of hepatotoxicity of menadione in rats.

The effects of induction of metallothionein (MT) on the toxicity of menadione were investigated in rat liver slices. The protective role of hepatic glutathione (GSH) was also studied and compared to that of MT. A 3-h incubation of rat liver slices with menadione (100-300 microM) containing medium (37 degrees C, pH 7.4, 95%O2:5%CO2) resulted in cellular toxicity, as shown by changes in cytosolic K, Ca and GSH concentrations and lactate dehydrogenase (LDH) leakage. A dose-dependent decrease in cytosolic K and GSH was observed concomitant with an increase in cytosolic Ca and LDH leakage after incubation with menadione. Pretreatment of rats with zinc sulphate (ZnSO4) (30 mg/kg body wt.) increased MT levels in liver slices and suppressed the toxicity of menadione. Intracellular GSH concentrations in liver slices were either depleted or increased by injection of rats with buthionine sulfoximine (BSO), (4 mmol/kg body wt.) and N-acetyl-L-cysteine (NAC) (1.6 g/kg body wt.), respectively. Intracellular GSH was found to be crucial in protection against menadione toxicity. Menadione toxicity was increased when the rats were injected with sodium phenobarbital (PB) (4 x 80 mg/kg body wt.). Pretreatment with Zn provided partial protection against menadione toxicity in liver slices from both BSO- and PB-injected rats. These findings suggest that induction of MT synthesis does protect against quinone-induced toxicity, but the role may be secondary to that of GSH. The mechanisms by which MT protect against menadione toxicity are still unclear but may involve protection of both redox cycling and sulphydryl arylation.


Glutathione disulfide reduction in tumor mitochondria after t-butyl hydroperoxide treatment.

Treatment of isolated mitochondria from rat hepatoma tumor cells (AS-30D) with the oxidant, t-butyl hydroperoxide (tBuOOH, 1 or 5 mumol/ml) resulted in the oxidation of glutathione (GSH to GSSG) and the formation of protein-glutathione mixed disulfides (ProSSG). The GSSG was retained inside of the hepatoma mitochondria. In the presence of ADP+succinate (5 or 10 mM), or ketoglutarate (10 mM) or malate (5 mM), the GSSG was reduced to GSH, but the amount of ProSSG stayed constant. With saline or ADP+glutamate (10 mM)/malate (0.1 mm) no reduction of GSSG to GSH occurred. The presence of antimycin (5 micrograms/ml) with ADP+succinate inhibited reduction. At a concentration of 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU, 0.5 mM) which inhibited a major portion of the glutathione reductase activity, the reduction of GSSG to replenish GSH was also inhibited. NADPH may play a critical role as well, for the addition of 2.4 mM NADPH to permeabilized hepatoma mitochondria fostered the reduction of GSSG after tBuOOH treatment. Therefore, hepatoma mitochondria possess a glutathione reductase-dependent system to reduce GSSG to GSH. The reaction only occurs with actively respiring mitochondria.


The effect of 2,2'-dichlorodiethyl sulfide on DNA synthesis of a murine stratified keratinocyte culture system.

A primary stratified keratinocyte culture resembling the epidermis in situ was used as a model for studying the effects of exposure to 2,2'-dichlorodiethyl sulfide, or sulfur mustard (SM), on DNA synthesis. A method that distinguishes between semi-conservative (s.c.) DNA synthesis and repair synthesis was used to determine if the former was inhibited following treatment with SM. In this method the density of the newly synthesized DNA was increased by incorporation of 5-bromo-2-deoxyuridine. Density gradient centrifugation was then used to isolate the heavy DNA for quantification. It was demonstrated that topically applied SM in the dose range of 1-10 nmole/cm2 inhibited s.c. DNA synthesis (replication) in a dose and time related manner. Inhibition of DNA replication by SM would result in inhibition of cell division which must be preceded by s.c. DNA synthesis. This failure to replace damaged germinative cells may lead to the destruction of the basal layer which is observed in vivo and in our epidermal culture following exposure to SM. This may also be related to development of vesication observed in exposed intact human skin.


Polycyclic aromatic hydrocarbon (PAH) ortho-quinone conjugate chemistry: kinetics of thiol addition to PAH ortho-quinones and structures of thioether adducts of naphthalene-1,2-dione.

Polycyclic aromatic hydrocarbon (PAH) o-quinones are products of an NADP+ dependent oxidation of non-K-region trans-dihydrodiols catalyzed by dihydrodiol dehydrogenase (EC 1.3.1.20). Since these PAH o-quinones could be detoxified by non-enzymatic or enzymatic conjugation with cellular thiols, their reactivity with 2-mercaptoethanol, cysteine and glutathione (GSH) was examined by ion-pair reverse phase high pressure liquid chromatography (RP-HPLC). Second-order rate constants for the addition of these thiols to naphthalene-1,2-dione (NPQ) in water ranging from 4.9 x 10(3) - 1.1 x 10(4) min-1 M-1 and the reactions were complete within 10 min. When these reactions were conducted at near physiological pH (50 mM potassium phosphate buffer pH 7.0), the rate constants increased by 2-orders of magnitude. When benzo[a]pyrene-7,8-dione (BPQ) was substituted in these reactions the second-order rate constants decreased by 2-3 orders of magnitude and the reactions took several hours to reach completion. The decrease in reactivity can be explained by the presence of the bay region in BPQ. Methylation influenced the reactivity of PAH o-quinones with GSH and the following order of reactivity was observed: 7,12-dimethyl-benz[a]anthracene-3,4-dione (7,12-DMBAQ) >> 12-methyl-BAQ, 7-methyl-BAQ and BAQ >> BPQ. Of these quinones 7,12-dimethyl-BAQ was almost equi-reactive with NPQ. This suggests that methyl substitution in the bay and peri regions enhances reactivity with GSH. Using NPQ as a model for other PAH o-quinones, N-acetyl-L-cysteine, L-cysteine and GSH conjugates of NPQ were synthesized and characterized by [1H]- and [13C]NMR. Evidence for Michael type 1,4-addition products was obtained in which the resultant adduct could exist as either a catechol or o-quinone. By contrast, L-cysteine was able to form adducts via S- or N-attack and N-attack gave a purple p-iminoquinone. There was no evidence for the formation of bis-N-acetyl-L-cysteinyl-, bis-glutathionyl adducts or phenolic coupled products. The toxicity of thiol conjugates of NPQ remains to be explored.


The reaction of melphalan with deoxyguanosine and deoxyguanylic acid.

The reaction of melphalan (phenylalanine mustard, I) with 2'-deoxyguanosine, followed by removal of the sugar in acid, yielded two products. The major product was identified as 4-(N-(2-guanin-7-ylethyl)-N-(2-hydroxyethyl)amino)phenyl- alanine (II) by ultra-violet absorption, mass and NMR spectroscopy. The minor product has already been identified as the corresponding bis-guaninyl adduct III (Tilby et al., Chem.-Biol. Interact., 73 (1990) 183-194). The reaction of melphalan with 5'-deoxyguanylic acid yielded the deoxyribonucleotide of II and products resulting from reaction with the phosphate group. The initial products, which were formed with a half-life of approximately 40 min at 37 degrees, still had a reactive chloroethyl group; this was displaced more slowly, by reaction with water or with another molecule of dGMP. The products of reaction of melphalan with DNA were released by treatment with acid (0.1 M HCl, 70 degrees, 30 min) and separated from each other on a cation exchange column. They were identified as II, III and an adenine adduct, in a ratio of approximately 3:1:2.


In vitro activation of heat shock transcription factor by 4-hydroxynonenal.

In the activation of eukaryotic heat shock genes, the acquisition of a binding ability to specific DNA sequence by a transcriptional activator, heat shock factor (HSF), is believed to be a crucial step. The induction of this new DNA binding activity of HSF is also obtained in a cell-free system (in vitro activation) by hyperthermia or at physiological temperature by calcium ions, low pH, urea, or non-ionic detergent. We report here the in vitro activation of HSF by treating at 0 degrees C a HeLa cell-free system with the aldehyde 4-hydroxynonenal (HNE), a highly cytotoxic product of lipid peroxidation. The in vitro activation of HSF by HNE occurred only if some components of the cell-free system were not sedimented at 100,000 x g. The reason for this is unclear but the release of active HSF from nuclei of unshocked cells and the involvement of Ca2+ contained in the mitochondria and ER have been excluded. Although HNE is known to be a sulfhydryl blocking agent, the results obtained with N-ethylmaleimide suggest that different mechanisms might be involved in the in vitro activation of HSF by HNE.


Synthetic studies of carbapenem and penem antibiotics. II. Synthesis of 3-acetyl-2-azetidinones by (2 + 2) cycloaddition of diketene and Schiff bases.

It was found that (2 + 2) cycloaddition reaction of diketene with Schiff bases was effectively promoted by imidazole as a catalyst to afford 3-acetyl-2-azetidinone derivatives 4. As an application of this new method, a practical asymmetric synthesis of 4 and its conversion into (3S,4S)-4-carboxy-1-(di-p-anisylmethyl)-3-[(R)-1-hydroxyethyl]-2- azetidinone, which is a key intermediate for the synthesis of carbapenem and penem antibiotics, were accomplished.


Synthetic studies of carbapenem and penem antibiotics. III. A synthesis of a key intermediate for 1 beta-methylcarbapenem.

We synthesized useful intermediates 5 and 6 for 1 beta- and 1 alpha-methylcarbapenems from 4-carboxy-3-[(R)-1-hydroxyethyl]-2-azetidinone 4 as a starting material by using stereoselective hydrogenation and hydroboration, respectively. A practical synthetic route from 4 to the (3S,4S)-4-[(R)-1-carboxyethyl]-3-[(R)-1-hydroxyethyl]-2-azetidinone derivative 1, a useful intermediate for the synthesis of 1 beta-methylcarbapenem antibiotics, was established.


Isolation and structures of antibacterial binaphtho-alpha-pyrones, talaroderxines A and B, from Talaromyces derxii.

New compounds designated talaroderxines A (1) and B (2) were isolated from a new heterothallic ascomycetous fungus, Talaromyces derxii, cultivated on rice. The structures of 1a and 1b were elucidated by means of spectroscopic examination and chemical reactions. Talaroderxines A (1a) and B (1b) are atropisomers of a 6,6'-binaphtho-alpha-pyrone derivative, and have strong antibacterial activity against Bacillus subtilis.


Preparation of three positional isomers of diglucosyl-cyclomaltohexaose.

Three positional isomers of diglucosyl-cyclomaltohexaose (diglucosyl-cG6) were chemically synthesized via 6(1),6(2)-, 6(1),6(3)-, and 6(1),6(4)-di-O-(TERT-butyldimethylsilyl)-cG6S (1, 2, and 3) prepared regiospecifically. Glucosylation of bis(2,3-di-O-acetyl)tetrakis(2,3,6-tri-O-acetyl)-CG6S obtained from the three regioisomeric compounds 1, 2, and 3 with 2,3,4,6-tetra-O-benzyl-1-O-trichloroacetimidoyl-alpha-D-glucopyran ose, followed by debenzylation and then deacetylation, afforded 6(1),6(2)-, 6(1),6(3)-, and 6(1),6(4)-di-O-(alpha-D-glucopyranosyl)-cG6S (10, 11, and 12) together with configurational isomers. The desired compounds 10, 11, and 12 containing two (1----6)-alpha-linkages were isolated from the mixtures of their configurational isomers by high performance liquid chromatography. The three diglucosyl-cG6S synthesized chemically were used as authentic samples to identify the components in a mixture of diglucosyl-cG6S produced by an enzymatic process.


Syntheses of a glycerophospholipid, C16-platelet activating factor and a palmitoyl analogue of M-5, an anti-inflammatory glyceroglycolipid.

From a chiral C4-epoxide (-)-3, which is one of the synthons in our synthetic strategy for complex lipids, a glycerophospholipid C16-platelet activating factor (C16-PAF, 1) and a palmitoyl analogue (2) of an anti-inflammatory glyceroglycolipid M-5, which was previously isolated from the Okinawan marine sponge Phyllospongia foliascens, have been synthesized.


Synthesis and antibacterial activity of some imidazo[1,2-a]pyrimidine derivatives.

A series of 75 imidazo[1,2-a]pyrimidine derivatives were synthesized. The "in vitro" antibacterial activity of these compounds and their corresponding alpha-bromoketones against a variety of gram (+), gram (-) bacteria and Mycobacterium species is reported. Some of the prepared derivatives exhibited potent antimicrobial activity.


Central cholinergic agents. III. Synthesis of 2-alkoxy-2,8- diazaspiro[4.5]decane-1,3-diones as muscarinic agonists.

A series of 2-alkoxy-2,8-diazaspiro[4.5]decane-1,3-diones and related compounds were synthesized and tested for muscarinic receptor binding affinity using [3H]pirenzepine and [3H]oxotremorine M as ligands. They were also evaluated for agonistic activities in the guinea pig ileum assay. 2-Methoxy- 2,8-diazaspiro[4.5]decane-1,3-dione (1i) was found to be a relatively M1 selective agonist. It reversed CO2-induced impairment of passive avoidance response with long duration of action, but also displayed peripheral effects at low doses. To minimize these side effects, we proposed the idea of conjugation of 1i with a muscarinic antagonist. The carbamate linked conjugate (1u) of 1i with methylatropine was therefore examined.


Synthesis of N-substituted C-normorphinans and their pharmacological properties.

Several N-substituted C-normorphinans (VIII and IX) were synthesized and tested for their analgetic and narcotic antagonist activities and physical dependence capacity. Treatment of N-formyl- octahydro-2-pyrindine (IIIc) with polyphosphoric acid readily gave N-formyl-C-normorphinan (IV). The N-nor bases (V and VII) obtained from IV were converted to VIII and IX. The N-methyl derivative (I), which was previously reported to be inactive by Haffner's method, exhibited potent analgetic activity by the hot plate method and the AcOH-induced writhing test. Compounds VIII and IX showed pharmacological properties similar to those of N-substituted morphinans and exhibited agonist (analgetic) and/or narcotic antagonist activities. The C-nor analogue (IXa) of cyclorphan (IIc) exhibited potent analgetic and antagonist activities with no physical dependence capacity in the single-dose suppression tests both in rats and monkeys.


Platelet activating factor (PAF) antagonists contained in medicinal plants: lignans and sesquiterpenes.

Hot aqueous extracts of medicinal plants were tested for their inhibitory effect on the binding of platelet activating factor (PAF) to rabbit platelets. The extracts of Forsythia suspensa VAHL. (Oleaceae), Arctium lappa L. (Compositae) and Centipeda minima (L.) A. BRAUN et ASCHERS (Compositae) showed significant activities. Since the main constituents of F. suspensa and A. lappa are lignans, 30 lignans were tested for their inhibitory effects on PAF binding to platelets and 9 lignans were found active. Four sesquiterpenes were isolated as active compounds from C. minima. In particular 6-O-angeloylplenolin and 6-O-senecioyplenolin are the most potent and specific PAF antagonists found in this study.


7-Oxodihydrokarounidiol [7-oxo-D:C-friedo-olean-8-ene-3 alpha,29-diol], a novel triterpene from Trichosanthes kirilowii.

The structure of 7-oxodihydrokarounidiol [7-oxo-D:C-friedo-olean-8-ene-3 alpha,29-diol], isolates from the seeds of Trichosanthes kirilowii (Cucurbitaceae), was determined by chemical correlation with karounidiol [D:C-friedo-oleana-7,9(11)-diene-3 alpha,29-diol] from the same source. Two other natural products, viz. bryonolic acid (3 beta-hydroxy-D:C-friedo-olean-8-en-29-oic acid) and bryononic acid (3-oxo-D:C-friedo-olean-8-en-29-oic acid), were also correlated with karounidiol.


Studies on the conformational aspects of inulin oligomers.

The conformational studies of inulin oligomers from G-F2 to G-F9, which isolated from Platycodon grandiflorum, suggested a plausible conformational change between G-F7 and G-F8 from the trends in their chemical shift patterns and molecular rotation; the oligomers higher than G-F8 would form some secondary conformations more rigid than shorter oligomers. On the other hand, spin-lattice relaxation (T1) studies of the protons proposed through-space interactions of 2- and 4-H's of glucose moiety in G-F5, presumably with some atom(s) of the terminal fructose moiety. This would reflect that the inulin molecule adopts a 5/1 helix.


Inhibitory effects of glycyrrhetic acid and its related compounds on 3 alpha-hydroxysteroid dehydrogenase of rat liver cytosol.

Glycyrrhetic acid (GA), aglycone of glycyrrhizin (GL), inhibited potently (I50 = 7 x 10(-6) M) and non-competitively the activity of NAD(P)+-linked 3 alpha-hydroxysteroid dehydrogenase of rat liver cytosol. The inhibition was slightly weaker than that of indomethacin, a potent anti-inflammatory agent, but stronger than that of dexamethasone, another anti-inflammatory agent. GL, GA monoglucuronide, and 3-epi-glycyrrhetic acid also inhibited this enzyme activity, but did so less effectively (I50 = 5-8 x 10(-5) M). Carbenoxolone (GA 3-hemisuccinate) and 3-keto-glycyrrhetic acid showed potent inhibitory effects similar to GA, and 18 alpha-GA showed the most powerful inhibition of the activity.


Biochemical studies on oral toxicity of ricin. V. The role of lectin activity in the intestinal absorption of ricin.

In order to investigate a possible role of lectin activity of ricin in its absorption from the small intestine, we prepared two ricin derivatives. BMH-ricin, prepared by crosslinking A and B chains of ricin with 1,6-bismaleimidohexane, was nearly non-toxic but the lectin activity was unaltered. And, NBS-ricin, prepared by the oxidation of tryptophanyl residues of ricin with N-bromosuccinimide, was not only non-toxic but also non-lectinic. After the oral administration of ricin derivatives to rats, their interaction with the digestive tract and absorption into the circulatory systems have been compared with those of ricin, immunochemically and histologically. It was shown by immunostaining that ricin and BMH-ricin could bind to the intestinal mucosa, whereas NBS-ricin could not. No appreciable damage in the small intestine from rats treated with either BMH-ricin or NBS-ricin has been observed, in contrast to ricin treatment where severe impairment of the small intestinal tissues resulted after 5 h. Immunoreactive ricin in the liver has been determined with the ricin enzyme immunoassay (EIA). When compared at 48 h after oral administration, NBS-ricin was not detected, whereas BMH-ricin was found to be 38 micrograms/liver and ricin 100 micrograms/liver. From these results, it was inferred that the lectin activity of ricin plays an important role in the absorption of ricin from the small intestine and that the absorption of ricin protein was enhanced by its high toxicity.


Conformation-dependent change in antitumor activity of linear and branched (1----3)-beta-D-glucans on the basis of conformational elucidation by carbon-13 nuclear magnetic resonance spectroscopy.

The antitumor activity of (1----3)-beta-D-glucans was tested in order to clarify its conformation-dependent response together with conformational elucidation by carbon-13 nuclear magnetic resonance (13C-NMR) spectroscopy. It was shown that the following three conformations, single chain, single helix and triple helix, are readily distinguished by the high-resolution solid-state 13C-NMR method. It turned out that preparations of linear (1----3)-beta-D-glucans of a triple helical conformation were ineffective in the inhibition of tumor growth. These linear (1----3)-beta-D-glucans were converted to an effective form in the inhibition of tumor growth when they were lyophilized from dimethyl sulfoxide (DMSO) solutions as a result of a conformational change from the triple helical to the single chain forms. They were not effective, however, when assayed in DMSO solution. In contrast, it was found that a branched (1----3)-beta-D-glucan is effective not only in either saline solutions of the triple helical sample or the lyophilized sample from DMSO, but also in DMSO solution. The aforementioned drastic change in antitumor activity was interpreted in terms of resulting conformational changes as analyzed by the 13C-NMR method.


Characterization of cationic acid phosphatase isozyme from rat liver mitochondria.

Acid phosphatase isozyme was highly purified from rat liver mitochondrial fraction. The enzyme showed an isoelectric point value of above 9.5 on isoelectric focusing, and the apparent molecular weight was estimated to be 32000 by Sephadex G-100 gel filtration or 16000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme catalyzed the hydrolysis of adenosine 5'-triphosphate, adenosine 5'-diphosphate, thiamine pyrophosphate, inorganic pyrophosphate, and phosphoprotein such as casein and phosvitin, but not of several phosphomonoesters, except for p-nitrophenyl phosphate and o-phosphotyrosine. The enzyme was not inhibited by L-(+)-tartrate, and was significantly activated by Fe2+ and reducing agents such as ascorbic acid, L-cysteine,and dithiothreitol. The enzyme was found to be distributed in various rat tissues including liver, spleen, kidney, small intestine, lung, stomach, brain and heart, but not in skeletal muscle.


Purification and characterization of gamma-enolase from various mammals.

The gamma subunit of enolase (gamma-enolase) was purified from the brain tissues of cow, dog, goat, pig, rabbit, and rat. The purification was achieved in only three steps: ammonium sulfate-precipitation, DE 53 cellulose ion-exchange chromatography, and polyacrylamide gel electrophoresis (PAGE) in a preparative mode. The purification procedure was comparatively more simple than previously reported methods, and the yield of gamma-enolase was sufficient for subsequent structural and immunological analyses. In all mammals, the purified gamma-enolase migrated in sodium dodecyl sulfate-PAGE (SDS-PAGE) with a molecular mass of 46 kilodaltons (kDa), and the immunological cross-reactivity between those gamma-enolases was very strong. The structural homology of these gamma-enolases was examined by peptide mapping using cyanogen bromide cleavage and subsequent two-dimensional electrophoresis. The resulting peptide patterns were highly similar and in cow, dog, and goat, the patterns were almost identical. These results indicate that structural homology, that is, the species non-specificity of gamma-enolase, appears to be very high.


Fluconazole: a potent inhibitor of cytochrome P-450-dependent drug-metabolism in mice and humans in vivo. Comparative study with ketoconazole.

The inhibitory effect of fluconazole (FCZ), a bis-triazole antimycotic, on mouse hepatic microsomal cytochrome P-450-mediated drug-metabolizing enzyme system was compared with those of ketoconazole (KCZ) in vivo and in vitro. Additionally, the change in the hepatic oxidative drug-metabolizing capacity in humans treated with FCZ was followed. The pentobarbital sleeping time in mice given a single dose of 1-10 mg/kg of FCZ or 30-50 mg/kg of KCZ was prolonged significantly, and the potency of FCZ for the prolongation of sleeping time was greater than that of KCZ. In contrast, in vitro the affinity and the inhibitory potency of FCZ for cytochrome P-450 and aminopyrine N-demethylation were 4- to 6-fold smaller than those of KCZ. However, the order of the inhibitory potencies among antimycotics for this enzyme systems in vitro was reversed by the addition of albumin into the reaction mixture. These results indicate that the difference in the plasma protein binding properties between FCZ and KCZ is an important factor which leads to a reverse in the order of their inhibitory potencies for this enzyme system in vitro and in vivo. The ratio of 6-beta-hydroxycortisol (6 beta-OHF) to cortisol (F) in urine, used as an indicator of oxidative drug-metabolizing capacity in humans, decreased to 50% of the original level during treatment with 200 mg/d of FCZ.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of cyclodextrins on biological membrane. II. Mechanism of enhancement on the intestinal absorption of non-absorbable drug by cyclodextrins.

The effects of two kinds of cyclodextrins (CyDs), alpha- and beta-CyD, on biological membranes were investigated by measuring changes in the absorption of a non-absorbable drug, sulfanilic acid (SA), from the rat small intestine, using in situ and in vitro experiments. After pretreatment with a mucolytic agent, N-acetyl-L-cysteine (N-Ac), only beta-CyD increased the absorption of SA significantly compared to the absorption without pretreatment. The mechanism of the enhancing effect of CyDs on the absorption of SA was discussed. Almost no morphological change in the small intestine was observed by pretreatment with N-Ac alone, N-Ac or alpha- or beta-CyD combinations. The liberation of membrane components differed among the CyDs, e.g., alpha-CyD selectively released phospholipid while beta-CyD released mainly cholesterol from the intestinal membrane. It is suggested that the interaction of membrane components with CyDs may be at least partly responsible for the enhanced absorption of SA. Moreover it was found from in vitro electrophysiological experiment, that the alteration in enhanced permeability caused by beta-CyD occurred primarily in the transcellular pathways, rather than in the paracellular pathways of the small intestine. These results suggest that the enhancement of intestinal absorption by beta-CyD, after removal of the mucin layer from the intestinal surface, is due to the interaction between the membrane components and CyD. This interaction would induce disorder in cell membrane lipid, resulting in the increased permeability of the transcellular route.


Interaction between polyethylene films and bromhexine HCl in solid dosage form. IV. Prevention of the sorption by addition of magnesium aluminum silicate.

The effects of magnesium aluminum silicate (MAS) addition on the sorption of bromhexine HCl to polyethylene film in tablets were studied. The addition of MAS prevented the sorption of bromhexine HCl to polyethylene film. In order to investigate the mechanism, the interaction between bromhexine HCl and MAS was studied by the powder X-ray diffraction method. It was observed that bromhexine HCl was preferentially adsorbed to the surface of MAS rather than to polyethylene film. The adsorption was accelerated at high temperature and reduced pressure conditions. The sorption of bromhexine base and bromhexine HCl to packaging material were compared using tablet dosage forms. The sorption of bromhexine base to polyethylene film was greater than that of bromhexine HCl.


An assessment of indomethacin-induced mucosal damage in vivo by measuring the metabolism of salicylamide in rabbit intestine.

Indomethacin-induced mucosal damage was assessed in vivo by measuring salicylamide (SAM) metabolism in rabbit intestine. Intestinal mucosal damage 48 h after oral indomethacin (500 mg/kg) administration was examined using a scanning electron microscope. Duodenal, jejunal and ileal mucosal toxicity was compared with that in controls. Intestinal first-pass metabolism of SAM was studied using in situ intestinal sacs with intact mesenteric venous blood collection. The appearance of both SAM and its metabolites in the mesenteric venous blood was measured following cannulation of the mesenteric vein of the exposed intestine and collecting all venous blood draining from the absorbing region. Following oral pretreatment with indomethacin, the appearance of SAM and SAM glucuronide (SAMG) in the mesenteric venous blood was significantly increased. The concentrations of SAM and SAMG in the blood increased following intraduodenal administration of SAM in vivo in rabbits orally pretreated with indomethacin compared with controls. However, after intravenous administration of SAM, the blood concentration of SAM and SAMG was not increased compared with controls. These findings suggest that the differences in intestinal first-pass metabolism of SAM may be due to the intestinal mucosal damage induced by oral indomethacin pretreatment. The results indicate that the alteration of intestinal first-pass metabolism of a marker compound may be utilized to assess intestinal mucosal damage in vivo.


Homeostasis as regulated by activated macrophage. VI. Protective effect of LPSw (a lipopolysaccharide from wheat flour) against acute infection by Toxoplasma gondii in mice.

An oral administration of partially purified LPSw, a lipopolysaccharide (LPS) from wheat flour, at a concentration of 20 ng/ml in drinking water beginning 1d after infection significantly decreased mouse mortality and prevented animal weight loss in acute infection with Toxoplasma gondii. Whereas 71% (5/7) of mice in a control group that did not receive LPSw died of toxoplasmosis, only 14% (1/7) of mice treated with LPSw died (p less than 0.05). The administration of LPS purified from Bordetella pertussis also significantly decreased the mortality of infected mice. LPS from Escherichia coli and synthetic lipid A (LA-15-PP(506)), however, did not show a significant decrease in mortality.


Homeostasis as regulated by activated macrophage. VII. Suppression of serum cholesterol level by LPSw (a lipopolysaccharide from wheat flour) in WHHL (Watanabe heritable hyperlipidemic) rabbit.

The effect of LPSw (a lipopolysaccharide from wheat flour) on cholesterol catabolism was examined using WHHL (Watanabe heritable hyperlipidemic) rabbit, which is an experimental model of familial hyperlipidemia. The serum cholesterol level of the animal decreased by the addition of LPSw to drinking water. Following cessation of the addition of LPSw to the drinking water, the cholesterol level was decreased for 30 to 40d and then gradually elevated. The serum level of apolipoprotein B, which is a constituent of apolipoprotein of low density lipoprotein (LDL), also decreased in accord with serum cholesterol at a nearly coincident rate. Conversely, the level of apolipoprotein A-I, which is a constituent of apolipoprotein of high density lipoprotein (HDL), did not change, nor did HDL-cholesterol. Furthermore, the atherosclerosis risk factor, expressed as the ratio of apolipoprotein B to apolipoprotein A-I, was decreased by LPSw administration.


Homeostasis as regulated by activated macrophage. VIII. LPSw (a lipopolysaccharide from wheat flour) can regulate bone resorption of chick embryo.

The effect of LPSw (a lipopolysaccharide from wheat flour) on the bone resorption of 18-d chick embryonic calvaria was examined in an organ culture following the method of Raisz. Bone was prelabeled in culture medium containing 45Ca and chased in a cold medium. On addition of test samples, labeled calcium was released indicating the grade of bone resorption. LPSw (10-100 ng/ml) stimulated bone resorption, showing an effect comparable to parathyroid hormone (PTH) (1 U/ml). PTH at 1 U/ml decreased the total amount of calcium and phosphorus, while LPSw did not. LPSw is thus assumed to stimulate bone resorption more actively than PTH.


Homeostasis as regulated by activated macrophage. IX. Enhancement effect of LPSw (a lipopolysaccharide from wheat flour) on hen egg-laying and breaking strength of eggshell.

Oral administration of LPSw (a lipopolysaccharide from wheat flour) given at 60 micrograms/hen/d in drinking water, markedly enhanced eggshell strength. The monthly percentage of eggs laid with a shell strength of more than 4 kg to the total number of eggs was 32% in the group given LPSw in drinking water while it was 12% in the control group given plain water. At the same time, LPSw caused a 30% enhancement of total monthly number of eggs laid over that of control.


Pharmacoepidemiological study on adverse reactions of antiepileptic drugs.

The relationship between the occurrence of side effects (SEs) and drug factors, such as antiepileptic drugs (AEDs), daily dose, duration of treatment, drug combination pattern, total and free serum concentrations, metabolite per parent level ratio as an index of metabolism ability, and co-medicated drugs except AEDs were evaluated in 227 outpatients with epilepsy. The possible influences of certain physiological and/or the pathophysiological factors were also evaluated. SEs with 19 clinical signs were observed in 66.1% of all patients. There was no definite dose- or serum concentration-dependent increase in the incidence of SEs. Stepwise discriminant function analysis revealed that benzodiazepines (BZN) polytherapy with AEDs produced a higher incidence of somnolence and general fatigue than did any other AED or drug combination. The effects of various drug combination patterns on the incidence of SEs were also evaluated on the basis of observed frequencies. The incidence of somnolence was significantly higher in patients taking phenytoin (PHT) plus carbamazepine (CBZ) therapy, and in patients taking BZN plus either PHT, phenobarbital (PB) or CBZ therapy compared with patients taking either PHT, PB or CBZ therapy. Other responsible drug combination patterns were PHT plus valproic acid (VPA) therapy for mental function impairment, acetazolamide (AZM) polytherapy with PB or PHT for dry mouth, and CBZ plus BZN therapy for constipation. In this study, the stratifying points (occurrence limits) of SEs were detected in various variables such as the number of prescribed drugs, daily dose and serum concentrations. Interestingly, these limits are within the commonly accepted "therapeutic range" or "usual daily dose," and some of these limits shifted down when another AED was co-medicated.(ABSTRACT TRUNCATED AT 250 WORDS)


Pharmacokinetics of [6]-gingerol after intravenous administration in rats with acute renal or hepatic failure.

The pharmacokinetics of [6]-gingerol were investigated in rats with acute renal failure induced by bilateral nephrectomy, or those with acute hepatic failure induced by a single oral administration of carbon tetrachloride (CCl4), to clarify the contribution of the kidney and liver to the elimination process of [6]-gingerol. After bolus intravenous administration, a plasma concentration-time curve of [6]-gingerol was illustrated by a two-compartment open model. There was no significant difference in either the plasma concentration-time curve or any pharmacokinetic parameters between the control and nephrectomized rats. It is suggested, therefore, that renal excretion does not contribute at all to the disappearance of [6]-gingerol from plasma in rats. In contrast, hepatic intoxication with CCl4 elevated the plasma concentration of [6]-gingerol at the terminal phase. Its elimination half-life increased significantly, from 8.5 to 11.0 min, in CCl4-intoxicated rats. The extent of [6]-gingerol bound to serum protein was more than 90% and was affected very slightly by the CCl4-intoxication. These aspects indicate that [6]-gingerol is eliminated partly by the liver.


Relationship between the effects on bacterial activity of selected disinfectants and the hydrophobic characters of dibasic acid diesters.

We prepared test solutions which contained 80% (v/v) ethanol and 0.2% (w/v) chlorhexidine (CH) or benzalkonium chloride (BC) with or without a dibasic acid diester. After complete evaporation of the ethanol from the solution on filter paper, an overnight broth culture (Staphylococcus aureus) was repeatedly inoculated onto the filter paper, and viable bacterial counts were measured at 5 min after the last inoculation. By comparison with viable counts for CH or BC alone, we estimated the potentiating effects of dibasic acid diester on the bactericidal activity of CH or BC, and confirmed that this activity of the two disinfectants was potentiated in the presence of certain compounds in the homologs of di-n-butyl esters of aliphatic dibasic acid, and di-alkyl esters of adipic and phthalic acid. Diisobutyl adipate, one of the most effective diesters, substantially enhanced the bactericidal activities of benzethonium chloride, cetyl pyridinium chloride and didecyl dimethyl ammonium chloride, as well as CH and BC, but not those of polyhexamethylene biguanide or alkyldiaminoethyl glycinate. The potentiating effects of dibasic acid diesters observed for both CH and BC seemed to be affected by the hydrophobic character of these diesters themselves and are also expressed well by a particular quadratic equation as a function of these characters: namely, capacity factors, as determined by high-performance liquid chromatography.


Purines. LII. Synthesis and biological evaluation of 8-methylguanine 7-oxide and its 9-arylmethyl derivatives.

The synthesis of 8-methylguanine 7-oxide (3) was accomplished via a "phenacylamine route", which started from condensation of alpha-(4-methoxybenzylamino)propiophenone (6), prepared by coupling of alpha-bromopropiophenone (4) and 4-methoxybenzylamine (5), with 2-amino-6-chloro-5-nitro-4(3H)-pyrimidinone (7) and proceeded through cyclization of the resulting phenacylaminopyrimidinone (8) and removal of the 4-methoxybenzyl group. The N-oxide 3 and its 9-arylmethyl derivatives 9 and 11 showed only very weak antileukemic activity and no antimicrobial activity.


Enzyme labeling in steroid enzyme immunoassays. The p-nitrophenyl ester method for alkaline phosphatase and glucose oxidase labelings.

The p-nitrophenyl ester method was assessed as an enzyme labeling technique. The active ester of a carboxylated testosterone derivative was treated with alkaline phosphatase and glucose oxidase to give labeled antigens, using various molar ratios of steroid to enzyme. Satisfactory immunoreactivities with an anti-testosterone antibody in an enzyme immunoassay system were obtained with the labeled antigens prepared at pH 8.5 by the use of molar ratios higher than 30 and 10, respectively, in the alkaline phosphatase and glucose oxidase labelings.