Reappraisal of the value of mode-A-echoencephalography.

The result of the first 425 cases studied in the Department of Neurology and Psychiatry, NTUH, by Mode A ultrasonic pulse techniques in the diagnosis of intracranial lesions is presented and compared with the findings in EEG, angiogram, air study and operation. The overall diagnostic accuracy in predicting midline echo shifting is 95.3%. The degree of echo shift is shown to be more closely related to the shift of venous angle than that of the anterior cerebral artery. The benefit of the combined use of echo and EEG is stressed. A method aiming at decreasing the operator bias is presented.


Age of onset and psychometric correlates of MMPI profiles in major motor epilepsy.

The purpose of the present study was to examine the MMPI performances of patients with major motor epilepsy and of nonepileptic brain damaged patients in relation to age of onset, duration of brain dysfunction and adequacy of intellectual functions. The present results suggest that MMPI indices of emotional disturbance are more closely associated with the degree of psychometric and cognitive impairment than they are with age of onset or duration of brain dysfunction. This effect is evident both in patients with major motor epilepsy and in nonepileptic brain damaged subjects.


Effects of changes in standing medication distribution times on PRN drug use.

Effects of changes in standing distribution times of medication on "as needed", prn, medication use on an inpatient psychiatric unit were studied. It was found that significantly less prn doses were distributed when standing medications were administred at 8 A.M., 12 Noon, 4 P.M., and 9 P.M. rather than at 10 A.M., 2 P.M., 6 P.M., AND 10 P.M. Data on three categories of prn drugs were collected. Of the three groups, significant effects were found for tranquilizers and hypnotics, but not for analgesics. The results are discusses in relation to psychosocial interactions on the unit and pharmacologic effects. The current psychopharmacology literature is replete with studies of drug efficacy and pharmacologic mechanisms of action. Less clinical investigation has been conducted on the effects of drug administration. Since 1958, when Tibbits first questioned the need to give chlorpromazine in divided standing doses, many clinical investigators have evaluated the effects of medication administration frequency on clinical outcome, Clinical trials of this type have been conducted for major tranquilizers and tricyclic antidepressants. A patient's complete medication schedule includes drugs given on a regularly standing basis and those given on a pro re nata (prn), according to the circumstances, basis. The effects of varying the times throughout the day at which standing medication doses are distributed have not been investigated. This study was designed to investigate the effects of alteration in administration schedules on the prn distribution of drugs in an inpatient psychiatric hospital setting.


An implementation of therapeutic community in a private mental health center.

The concepts of milieu therapy have, in the past 20 years, been widely applied to various facilities for treatment of people with emotional and mental illness. One type of milieu therapy, the Therapeutic Community, has gained increasing acceptance, as a means of implementing milieu therapy. This paper describes the application of therapeutic community concepts to the in-hospital population of a private, open-staffed, open-door psychiatric facility, and some of the effects that this has had on both the staff, and the patient population. Since the development of the therapeutic community concept almost 25 years ago, programs have been developed in various centers which implement the concepts of social psychiatry in different ways. This paper describes the development of a particular therapeutic community on the in-patient adult and adolescent psychiatric services of a private psychiatric hospital. Parkwood was developed as a small (40 bed) psychiatric hospital in Atlanta, Georgia in 1966 by a group of physicians wanting to implement the ideas of milieu therapy in an area where no such program was available. In its initial stages, the hospital -- of attractive, middle-class decor -- was built in a wooded setting on the edge of a large metropolis. It was open-staffed, with psychiatrists having an eclectic, though psychoanalytically based, orientation toward treatment. The concepts of milieu therapy were interpreted to mean the following: a warm, pleasant atmosphere, in  pleasant surroundings, conducive to the usual brief psychotherapy, chemotherapy, or somatic therapy, which had previously been used in non-milieu psychiatric hospitals. The total hospital patient population was divided in half and met in group sessions with a psychiatrist once each week. The Medical Director, a psychiatrist, had weekly staff meetings, in which he discussed various administrative problems. In 1969, a Medical Director with a therapeutic community orientation was employed. Over a five-year period, the total complexion of the hospital changed to its present state, an open-staffed, open door, comprehensive, community mental health center financed on a private basis. The facility now provides out-patient, partial or total hospitalization, emergency services, and community education for children, adolescents, adults and alcohol addicted patients. This paper deals with the therapeutic community on the adult and adolescent units.


Preoccupation with death in schizophrenic men.

A systemic review of psychiatric histories of 205 hospitalized, schizophrenic men (veterans) revealed that 80 had expressed fear of dying, being killed, of no longer existing, or other evidence of preoccupation with death. These expressions of fear appeared either at the clinical onset of the psychosis, or during exacerbations, and often coincided with schizophrenic panic. The similarity of expressions of the fear among the patients was striking, and they were practically indentical from one exacerbation to the next. This consistency suggests that the fear of death stems from schizophrenic psychopathology rather than from individual situational influence. The explicitly stated preoccupation with death was quite common in this sample (40%), and contrary to expectation, was not limited to cases with favorable outcome.


Primary intracranial hypotension: the syndrome of spontaneous low cerebospinal fluid pressure with traction headache.

A case of primary Intracranial Hypotension (PIH) is described. This syndrome consists of spontaneous lowering of CSF pressure with traction headache. It is a benign disorder and is self-limited. The cause is unknown, but local choroid plexus vasospasm, possibly due to a hypothalamic disturbance, has been suggested, as has leakage of CSF through spontaneous arachnoid tears. Transient symptomatic relief was achieved with 5% CO2 inhalation, apparently due to increase in cerebral blood flow.


[How dangerous is lumbosacral myelography with water-soluble contrast media? (author's transl)].

In the exact diagnosis of space-occupying processes of the lumbosacral spine myelography with water-soluble contrast media is unsurpassed. Neurotoxicity of the water-soluble contrast media is known and feared in many centres. However, after 2500 exclusively in-patient lumbosacral myelographies and after a prospective study of 100 patients we were able to demonstrate that this investigation is only slightly dangerous.


[The importance of cytodiagnosis of perbronchial fine needle aspiration of mediastinal or hilar tumours (author's transl)].

In almost all bronchoscopy patients perbronchial puncture of hilar lymph nodes was performed routinely. There were 1883 punctures performed in 1841 patients. In histologically proven malignant processes tumour cells were demonstrated in the smear in 84% of the cases, whereas in 97 cases of sarcoidosis a cytological diagnosis could be made in 94%. On account of the high specificity, the harmlessness of the procedure and the minimal stress caused to the patient in comparison with mediastinoscopy or thoracoscopy perbronchial fine needle asperation is highly recommended in unclear mediastinal processes as well as to determine the operability of a bronchial carcinoma or stage of a tumour according to the TNM system.


[Successful treatment of a supraventricular re-entry tachycardia in a patient with WPW and sick-sinus syndrome with permanent rapid atrial and QRS-inhibited ventricular pacemaker stimulation (author's transl)].

A supraventricular re-entry tachycardia was successfully treated in a patient with WPW and sick-sinus syndrome by permanent rapid atrial pacing and ventricular demand pacing. The use of a demand pacemaker with bipolar electrode avoided slow ventricular rates resulting from second degree A-V block during rapid atrial pacing. The trigger system of this pacemaker was not disturbed by unipolar atrial stimulation. Ventricular rates responded satisfactorily to physical and to emotional stress as a result of improved A-V conduction in these circumstances. Rapid atrial pacing has prevented the re-entry tachycardia. In an emergency the atrial pacemaker can be inhibited by a magnet, but otherwise discharges permanently at a constant rapid rate.


[Exercise testing and atrial pacing in the diagnosis of latent coronary insufficiency (author's transl)].

Right atrial pacing at different rates was performed on 15 women and  11 men with persistent angina with normal resting and exercise ECG. In 21 subjects there developed during bicycle exercise unequivocal S-T depression of myocardial ischaemia at maximal frequency. In 16 of them the diagnosis was confirmed by rate-dependent attacks of angina. These results demonstrate that, especially in the elderly, the maximal rate (+/-15%) thought to give a full load to cardiovascular functions is not suitable for diagnosing with certainty coronary insufficiency on exercise. In such patients rate-stimulation test may help in recognising the basic disease. On the other hand, in younger patients, capable of marked increase in heart rate on maximal exercise, rapid atrial stimulation has no diagnostic advantage. This test is indicated in those subjects who during ergometric exercise have an insufficient increase in heart rate because of a general reduction in exercise tolerance.


[Post-traumatic and post-operative paresis of the leg plexus (author's transl)].

Post-traumatic and post-operative leg-plexus paresis is often misdiagnosed as sciatic or femoral paresis and thus falsely thought to be rare. In 20 post-traumatic cases the cause was predominantly or exclusively due to sacral plexus damage, with paralysis of the muscles innervated by the sciatic nerve and of the gluteal muscles. The most frequent cause was severe pelvic trauma with ring fracture of the pelvis, separation of the symphysis and iliosacral gap or fracture of the hip joint. Postoperative leg-plexus paresis was noted in eight cases after total hip-joint replacement (predominantly damage to the lumbar plexus) and twice after operatively treated fracture of the acetabulum. Both pressure and tearing lesions can be the basic cause.


[The management of a case of congenital erythropoetic prophyria (author's transl)].

The present condition of a now 18-year-old boy with congenital erythropoetic porphyria is reported. The case had been first described 15 years ago in this journal (Heilmeyer, L., et al.: Dtsch. med. Wschr.88 [1963], 2449). Physical and mental development has been normal. He has been on a maintenance dose of beta-carotene, 50 mg daily, for the last four years except during the winter months. This has allowed him to be in the open air in normal clothing. At the same time, the improvement in light tolerance has enable him to pursue normally the work of a biology laboratory technician. The genetic features are such that neither marriage nor children need be advised against.


[Children of mothers with phenylketonuria (author's transl)].

Microcephaly and considerable motor and mental retardation occurred in two non-phenylketonuric children of an untreated mother with phenylketonuria. The cerebral damage of the children must be considered the consequence of the maternal metabolic disorder. Since the first phenylketonuric children who were treated on strict diet are now reaching the age of marriage and pregnancy, the problem of maternal phenylketonuria is becoming topical. Published reports indicate that of 72 well documented cases with a maternal phenylalanine level above 200 mg/1 (1210 mumol/1) 39 offspring had microcephaly, in 33 intra-uterine growth had been retarded and in 25 there are cerebral palsy and seizures. Almost all had mental retardation. At the same time there have been reports about three normal children whose mothers had kept to a phenylalanine-low diet during their pregnancy.


Management of cystitis in young women.

The common clinical problem of recurrent symptomatic urinary tract infections (UTI) in young women may be inconvenient and distressing to both the patient and her family, particularly if the infections are not managed in a rational manner. A suggested approach is included in table 1.


Effect of inorganic iodide on thyroglobulin hydrolysis in cultured thyroid glands.

The process of thyroglobulin hydrolysis in mouse thyroid glands labelled in vitro was studied from 2-24 h after they had been maintained in tissue culture. The culture medium was supplemented with mononitrotyrosine to prevent deiodination of iodotyrosines. Hydrolysis of labelled thyroglobulin, under these conditions, led to the release of labelled iodotyrosines and iodothyronines. The rate of formation of these compounds was measured as an index of thyroglobulin hydrolysis (TH). TH was markedly stimulated by TSH. NaI inhibited TSH stimulation of TH at a concentration of 10(-5)M or greater. NaI, at similar concentrations,also markedly diminished or abolished the incorporation of 131I into thyroidal proteins from radioiodide-supplemented media. The addition of various inhibitors of iodination effectively blocked the effect of iodide on TH. In experiments where radioimmunossay was used to measure medium hormone concentrations, the release of unlabelled thyroxine (T4) and triiodothyronine (T3) induced by TSH was found to be significantly decreased in the presence of 10(-4)M NaI. These studies demonstrate that iodide inhibits the hydrolysis of thyroglobulin at or near concentrations which also inhibit iodination of thyroidal proteins. The present data suggest that formation of an iodinated compound is necessary for the effect of iodide. In addition, these studies demonstrate the utility of this in vitro system for the investigation of thyroid physiology.


Effect of acute potassium loading on plasma renin and on urinary aldosterone in rats.

The disposition of aldosterone radiometabolites in rats has been studied following iv [3H]aldosterone administration. Of injected [3H]-aldosterone, 0.31% is recovered in 24 h urine as free aldosterone and 0.08% as acid-labile conjugate. A simple, sensitive and reliable radioimmunoassay of free aldosterone has been developed and the effect of acute oral potassium loading (171, 513 or 769 mueq of KCl/100 g body weight) on 4 h aldosterone excretion, plasma renin concentration and sodium and potassium balance has been investigated. There was a positive correlation between log urinary aldosterone and potassium load (r = 0.92, P less than 0.001). Potassium induced a natriuresis which was correlated directly with the dose of potassium administered (r = 0.89, P less than 0.001). Ptasssium loading also increased plasma renin concentration which was correlated with the sodium excretion rate (r = 0.64, P less than 0.01). Prevention of a negative sodium balance during the 769 mueq potassium load was obtained by administration of 513 mueq sodium. In this experiment, plasma renin concentration increased little, whereas the aldosterone excretion rate was as high as during the 769 mueq potassium load without sodium addition.


Stimulation of gastrin secretion in the pig by parathyroid hormone and its inhibition by thyrocalcitonin.

Gastrin secretion was studied in 16 young anesthetized pigs weighing 14-26 kg. Test substances were infused (0.1 ml/min x 10-20 min) directly into the gastric antrum via a catheter in the right gastroepiploic artery. Samples were collected from a catheter in the right gastroepiploic vein and plasma gastrin was measured by radiommunoassay. The following results were observed: 1) basal gastrin in antral venous blood was 10-5 times that in peripheral blood (620+/-222 pg/ml vs. 41+/-10 pg/ml, 2) native bovine parathyroid hormone (PTH) and synthetic human 1-34 PTH (0.02-4U/min) produced rapid (within 10-30 min) and pronounced (approximately 10-fold) increases in gastrin release with no increase in plasma calcium and, in several animals, in the face of a falling plasma calcium concentration, 3) neither acute thyroidectomy nor infusion of porcine thyrocalcitonin (TCT), 0.5-2.5 U/min) consistently altered basal gastrin secretion (N=3-6), and 4) infusion of TCT (0.5 U/min)along with PTH (2U/min) significantly suppressed the 10-11-fold increase in gastrin release observed when PTH subsequently was infused alone in each pig (N=6). The results demonstrate that PTH can stimulate gastrin secretion in the pig and that TCT can suppress this effect.


Arginine vasopressin secretion in thyroidectomized sheep..

Indwelling, exteriorized, jugular vein catheters were placed in five thyroidectomized ewes at a time when myxedema was manifested clinically and chemically and three euthyroid sheep were used as controls. Post-operatively, tracer doses of [125I]-iodovasopressin were injected and serial blood specimens were obtained for determination of volume of distribution, plasma disappearance, and blood production rates. Serum vasopressin was measured by radioimmunoassay. The mean volumes of distribution for vasopressin in the hypothyroid and euthyroid sheep, respectively, were 8.15 and 5.90 liters, mean t1/2 of vasopressin 9.5 and 19.3 min, mean serum vasopressin concentrations 5.1 and 1.2 muU/ml, and mean blood production rates 2.84 and 0.23 mU/kg/h. Renal and organ biologic effectiveness of the elevated vasopressin levels was suggested by the lowered serum osmolalities in the hypothyroid sheep over controls (272 vs. 301 mosmol/kg). These results suggest an augmented secretion of vasopressin in the myxedematous state.


Serum 5alpha-androstane-3alpha,17beta-diol, androsterone, and testosterone concentrations in the male rat. Influence of age and gonadotropin stimulation.

Serum concentrations of testosterone plus dihydrotestosterone (T-DHT), 5 alpha-androstane-3 alpha, 17 beta-diol (Diol) and androsterone were measured during sexual maturation in male rats. Diol concentrations of 1 to 2.25 ng/ml were found in animals 10-90 days of age with no significant changes. Diol was the major androgen (5.75-8 times T-DHT) from age 20-40 days. Androsterone rose to 1.25 ng/ml at 25 days of age and declined to values of greater than 0.5 ng/ml from age 30-90 days. Testosterone-DHT levels were 1 ng/ml or less from 10-40 days of age, and then rose to a peak at 60 days. The ratio of Diol toT-DHT was significantly elevated from age 20-35 days, indicating that Diol is the major androgen in circulation at this time. Acute treatment of 33 day old rats with LH, but not with FSH, resulted in a dose-dependent increase in serum T-DHT, Diol, and androsterone. The dose-response and time course of response for the three steriods were nearly identical. Changes in testes capacity to secrete androgens were assessed 25, 33, 40 and 60 days of age by administering a maximum dose of LH before blood collection. Maximum response (sum of androsterone, Diol and T-DHT) occurred at 40 days with no further increase at 60 days.


Luteinizing hormone secretion and female lordosis behavior in male pigs.

Serum concentrations of luteinizing hormone (LH) and testosterone were monitored during the first 9 weeks of life in male pigs. In boars, testosterone was constant during this period, and LH was elevated from birth through day 21 and declined between days 21 and 28 to levels observed in adult boars. Castration on day 3 reduced testosterone concentrations without having an immediate effect on LH, but after day 21, LH was consistently greater in castrated males than in boars. Responsiveness of the early postnatal testes to LH was shown by reduced testosterone concentrations after treatment with LH-antiserum on day 2. Norethindrone had no effect on LH or testosterone when given on day 2 but decreased the concentrations of these hormones in 5-month-old boars. Similarly, estrogen had no effect on LH secretion in 4-day-old boars. In 9-month-old castrated males, estrogen decreased LH concentrations during the first 48 h of daily treatment, but after this, LH increased and returned to pretreatment levels by 88 h. Also, daily estrogen treatment of castrated males induced female lordosis behavior. From these observations we conclude that negative feedback control of LH secretion is inoperative in neonatal male pigs and that exogenous estrogen can induce responses characteristic of females in castrated postpubertal males.


Effect of thyrotropin on ornithine decarboxylase and of polyamines on RNA polymerase in the thyroid.

Two major forms of RNA polymerase were partially purified from calf thyroid nuclei. Type I showed greatest activity in 30 mM(NH4)2SO4, transcribed native DNA best and was resistant to the inhibitor alpha-amanitin. Type II showed greatest activity in 100 mM(NH4)2SO4, transcribed denatured DNA template best and was sensitive to low concentrations of alpha-amanitin...


Relation of triiodothyronine and reverse triiodothyronine administration in rats to hepatic L-triiodothyronine aminotransferase activity.

The effects of administration of 3,5,3'-triiodothyronine (T3) to normal and to hypothyroid male rats upon the hepatic activity of L-triiodothyronine aminotransferase were determined using 3,5-dinitro-L-tyrosine as substrate in the assay. Initial studies in normal rats demonstrated that basal enzyme activity was highest in liver and kidney of the organs tested, and that virtually no activity was detectable in skeletal muscle, serum, thyroid or pituitary gland. Hepatic enzyme activity increased from birth to a peak at 80-120 days and declined thereafter. Daily administration of T3 to normal rats in doses of 5 mug/100g BW for 8 days significantly elevated hepatic enzyme activity above normal. In daily doses of 2.5mug/100g BW, T3 restored the depressed enzyme activities in hypothyroid rats to normal. Daily administration of 3,3',5'-triiodothyronine (reverse T3) to normal rats in doses of 17.5 mug/100g BW and greater for 3 days increased L-T3 aminotransferase activity more than 30% above normal levels. Reverse T3 appeared to be approximately as active as T3 in increasing the hepatic activity of L-T3 aminotransferase.


In vivo inhibition of thyroid secretion by indomethacin.

Indomethacin (Ind) was administered to adult female rats to reduce endogenous prostaglandin (PG) synthesis in order to investigate the role of PGs in thyroid hormone secretin. This treatment markedly reduced thyroidal PGF levels (667.7 vs. 1822.1 pg/my, P less than .001). Although the plasma TSH concentrations were normal in the Ind-treated group (41.14 vs. 40.01 mug/100 ml), dramatic decreases were observed in plasma T3 (24.5 ca. 6.7 nf/100 ml, P less than .001) and T4 (5.6 vs. 0.7 mug/100 ml, P less than .001) levels...


Changes in hepatic steroid metabolism in rat following transplantation of four different clones of pituitary tumor cells.

The effect of four different clonal pituitary tumors on hepatic steroid metabolism was studied in male and female rats and this effect was related to the secretion of prolactin, LH and FSH from the tumors. Tumors derived from the cell line C811RAP caused an alteration of the steriod metabolism to a completely female pattern in male rats whereas it had little effect on metabolism in female rats. This tumor cell line was the only one which appeared to secrete prolactin according to the observed level of this hormone in host serum. Furthermore, extract from C811RAP tumor tissue increased the 5alpha-reductase activity in cultured HTC CELLS. The other cell lines tested (C29RAP, C13RAP and C311RAP) did not appear to secrete prolactin but did have general effects on the hepatic steroid metabolism of both sexes. These effects could not be interpreted as changes towards a female or a male pattern of metabolism. When extract from C13RAP or C211RAP tumor tissue was assayed in the HTC cell system, small or insignificant effects on the 5alpha-reductase activity were observed. In conclusion, it appears that the C811RAP pituitary tumor cell line produces a factor(s) that in duces a female type of hepatic steroid metabolism in the rat. This factor does not seem to be produced by the other cell lines tested. Based on the clonal nature of the cells and the fact that the C811RAP cells also secrete prolactin, it is reasonable to suggest that the mammotrophs of the normal female pituitary are the cells that produce this factor.


Parallel shift in circadian rhythms of adrenocortical activity and food intake in blinded and intact rats exposed to continuous illumination.

Twenty-four hour patterns of plasma corticosterone levels and food intake were determined at 3-4 h intervals weekly for 7 consecutive weeks in blinded and intact female adult rats exposed to continuous illumination. Plasma samples were obtained from the tail vein. Resting levels and circadian periodicity of plasma corticosterone levels determined by this method were essentially similar to those obtained by decapitation. Circadian periodicity of adrenocortical activity and food intake shifted in both blinded and intact rats. In each group, the patterns of shift in these two activities were essentially similar. Between the two groups, however, the patterns of shift were markedly different. In blinded rats under either constant light or regular light-dark alternation a fairly constant phase shift was observed after the 3rd week and phase reversal of rhythms was found at the end of the 7th week. An additional determination of 48 h pattern of plasma corticosterone levels at the end of the 7th week demonstrated a clear periodicity in the majority of the blinded rats when the patterns were determined individually. On the other hand, in the intact rats exposed to constant light the parallel shift in the two activities was observed for only two weeks. Although a significant difference between the peak and trough values was observed up to the 6th week in food intake and the 4th week for corticosterone levels, a plot of the plasma corticosterone levels determined at 4 h intervals for 48 h did not show any periodicity in individual rats at the 4th and 7th week. In the case of partial food deprivation (feeding time, 1900-2100 h), peak elevation of plasma corticosterone levels occurred at 1900 h for two consecutive weeks in the intact rats exposed to continuous illumination. These results indicate that rhythmicities of adrenocortical activity and food intake persist longer in the blinded rats than in the intact rats kept under constant light. The fact that the phase shift of adrenocortical activity was prevented by the artificial fixation of the feeding period indicates that the shift of adrenocortical activity is a secondary phenomenon to the shift in the periodicity of food intake.


The importance of thyroglobulin structure in thyroid peroxidase-catalyzed conversion of diiodotyrosine to thyroxine.

We have previously demonstrated that thyroid peroxidase (TPO) not only catalyzes the iodination of thyroglobulin and other proteins, but that it also catalyzes the intramolecular conversion of DIT residues to T4 (coupling reaction). The present study was designed to determine whether the native structure of thyroglobulin contributes to the efficiency of TPO-catalyzed coupling. Two lines of evidence are presented in support of the view that the conformation of thyroglobulin is important for TPO-catalyzed coupling. The first was based on comparison of T4 yields in thyroglobulin and other proteins. The second involved the effect of guanidine pretreatment on T4 yields in thyroglobulin. Both types of experiment provided evidence that the native structure of thyroglobulin contributes to the efficiency of the coupling reaction. Specificity of thyroid peroxidase activity, on the other hand, does not appear to be of importance in the coupling reaction.


Mechanisms of decreased insulin responsiveness of large adipocytes.

We have studied glucose metabolism using large adipocytes isolated from older, fatter rats (greater than 12 months old, greater than 550 g), and smaller cells obtained from younger, leaner animals (4-5 weeks old, 126-160 g). 2-Deoxyglucose uptake was equal in large and small adipocytes, while insulin mediated oxidation of [1(-14)C]glucose was greatly diminished (7-fold) in large cells. Thus, the defect in oxidation of the number one carbon atom of glucose (pentose pathway oxidation) is distal to the 2-deoxyglucose uptake system. However, this intracellular defect is not present in all pathways of glucose oxidation as demonstrated by the finding that [6(-14)C]glucose oxidation was comparable in small and large adipocytes. Thus, the number six carbon atom of glucose is oxidized normally indicating that glycolytic and Krebs cycle activity is intact in the large adipocyte. Furthermore, in large adipocytes conversion of glucose to total lipid was normal in the basal state and moderately decreased at high glucose concentrations in the presence of insulin (up to 35%). When the radioactivity in total lipids was fractionated, a severe decrease in glucose incorporation into fatty acids was found in the large cells. Total glucose uptake was also measured, and found to be 10-50% decreased in large cells, suggesting that the decreases in pentose pathway glucose metabolism and conversion to fatty acids lead to accumulation of free intracellular glucose with glucose efflux and a decrease in net glucose uptake. Comparing the 2-deoxyglucose uptake and glucose oxidation data showed that insulin promotes [6(-14)C]glucose oxidation by stimulating the processes responsible for 2-deoxyglucose uptake whereas insulin promotes [1(-14)C]glucose oxidation both by increasing these processes and by increasing the activity of the C-1 oxidative pathway. 1) the 2-deoxyglucose uptake system of the large adipocyte is basically intact, 2) [1(-14)C]glucose oxidation is markedly decreased in large adipocytes, while [6(-14)C]glucose oxidation is normal, and 3) in comparing small and large adipocytes, it appears that it is the ability of insulin to enhance glucose oxidation via the pentose pathway and to promote glucose incorporation into fatty acids which is most impaired in large adipocytes.


Lactogenesis in explant cultures of mammary tissue from pregnant cows.

The hormonal requirements for lactogenesis were investigated using explant cultures of mammary tissue obtained from cows at 30-40 days prepartum. Hormones used were insulin, hydrocortisone, and prolactin, and parameters examined were radioactive acetate incorporation into fatty acids, secretory response ratings, and histological and ultrastructural analysis. Data indicated that insulin was essential for mammary epithelial cell survival, but insulin alone did not result in the intiation of milk synthesis. The culture of explants in a medium containing insulin plus hydrocortisone resulted in alterations in the cytology of alveolar cells but no induction of milk synthesis. Biosynthesis results and secretory response ratings indicated that the initiation of milk synthesis occurred when explants were cultured in insulin and prolactin; however, synthesis was limited and alveolar integrity was not well maintained. Results from all parameters demonstrated that the maximal lactogenic response was obtained when the culture medium contained insulin, hydrocortisone and prolactin. After 48 h of culture in this medium, the rate of acetate incorporation into fatty acids had increased 3-fold and the markedly distended alveolar lumina contained many fat droplets and abundant eosinophilic staining secretion. However, an unusal amount of casein-like micelles and especially lipid also accumulated in the alveolar cells. The accumulation of milk components within the epithelial cells may be related indirectly to the accumulation of products in the lumina or perhaps related to a difference in the hormonal requirement between the initiation of milk synthesis and initiation of milk secretion.


Factors affecting cytidine uptake and utilization by the rat uterus.

The influence of cytidine dosage on uptake and distribution of [3H]cytidine by the uterus of the adult ovariectomized control rat was examined 20 min after injecting the nucleoside. Intracellular accumulation of free cytidine was observed only at a dosage of 4.2 mumol/100g. At lower dosages, the nucleoside was phosphorylated as rapidly as it entered uterine cells, thereby preventing its accumulation within cells. [3H]Cytidine (4.2 mumol/100 g) was used in time-course studies in ovariectomized control and estradiol-treated rats. From this study the following differences between uteri of control and estradiol-treated animals were noted: 1) both nucleotides and RNA were five to six times more highly labeled in treated than control animals after a five minute labeling period, 2) this difference between control and treated animals was reduced to twofold when the labeling period was extended to 20 min, 3) free nucleoside had begun to accumulate intracellularly in treated animals, but had not equilibrated with uterine extracellular space in controls 5 min after [3H]cytidine injection, and 4) at 20 min, the free nucleoside of the uterus was labeled about equally in control and treated rats. It is concluded that estradiol causes a rapid equilibration of circulating cytidine with the uterus and that increases in labeling of RNA and precursor nucleotides 2 h after hormone treatment are largely a consequence of this rapid equilibration.


Effect of lesions in the ventral noradrenergic tract produced by microinjection of 6-hydroxydopamine on gonadotropin release in the rat.

Noradrenergic innervation to the hypothalamus is provided principally by the ventral noradrenergic tract (VNAT) which carries the axons of noradrenergic neurons whose cell bodies lie in the brain stem. To determine the importance of the VNAT in the stimulation of LH release induced by progesterone in ovariectomized, estrogen-primed rats, 6-hydroxydopamine (6-OHDA), an agent which selectively destroys catecholaninergic neurons, was microinjected bilaterally into this tract, 2 days after priming of the ovariectomized animals with 5 mug of estradiol benzoate, sc. Following microinjection, 2 mg of progesterone was injected sc to provoke LH release. A surge of FSH and LH release occurred 6 h after progesterone in control animals and those injected with the ascorbic acid diluent into the VNAT. Injections of 6-OHDA into the tract completely blocked both the LHP AND FSH surge. Control injections of 6-OHDA into the superior colliculus, the caudate-putamen or the frontal cortex did not alter the release of FSH and LH induced by progesterone. Twenty-four hours after injection of 6-OHDA into the VNAT, there was a slight reduction in norepinephrine content in the anterior hypothalamic area and a significant reduction in the region of the arcuate nucleus and median eminence. In this experiment, 6-OHDA injections into the VNAT blocked not only the FSH and LH release induced by progesterone but also the increase in serum prolactin which was present in control animals. In normal females, injections of 6-OHDA into the VNAT blocked the proestrous discharge of LH and partially blocked that of FSH. It is concluded that acute interruption of the VNAT induced by a 6-OHDA can block not only the stimulation of FSH and LH induced by progesterone, but also the preovulatory discharge of gonadotropins. The results suggest that increased impulse traffic in the VNAT on the afternoon of proestrus may be involved in induction of the proestrous gonadotropin surge.


Identification of aromatase in the reptilian brain.

The present study tests the hypothesis that brain aromatase is an "ancient" property of nervous tissue and may be identified in homologues of the limbic system in a non-mammalian vertebrate, the turtle Chrysemys picta. Tissue homogenates (180 mg wet weight/2 ml) were incubated with [7 alpha-3H]androstenedione and cofactors for 60 min at 37 C. Estrone (E1) was isolated and characterized by thin layer chromatography, methylation and recrystallization to constant specific activity. No estradiol-17 beta was detected. Aromatase was found only in the forebrain but was diffusely distributed throughout this major brain division. No other neural or non-neural tissues, including mid- and hindbrain structures, testis, and ovary, synthesized detectable quantities of E1 in our system. The strio-amygdaloid complex of both sexes synthesized more E1 per unit weight than the preoptic-hypothalamic area (POA-HTH) or other forebrain structures. It is possible that the conversion of androgen to estrogen has biological significance in this species since the reaction occurred throughout the physiological temperature range experienced during activity in nature and sex differences in brain aromatase activity during the breeding season were apparent. These experiments in Chrysemys demonstrate that the synthesis of estrogen from androgen by the brain is not limited to mammals, but also occurs at a more primitive level of phylogenetic development. Restriction of aromatase to forebrain structures of the turtle is consistent with the neuroanatomic distribution of enzyme activity in the limbic system of mammals. Estrogen yield from adult turtle brain incubates (3.2-22.6 pmol/g) is more like that reported for fetal (2.7-33 pmol/g) than for adult (0.1-1.9 pmol/g) mammals. We suggest that the in situ synthesis of estrogen by the central nervous system has its orgins early in vertebrate evolution and may be a primitive characteristic of brain-steroid interactions that regulate physiological and behavioral sex in vertebrates.


Prolactin activation of phospholipase A activity in membrane preparations from mammary glands.

The addition of prolactin to the 150,000 x g sedimented fraction of mammary gland homogenates increased by about two-fold the rate of [3H]-arachidonic acid released from phosphatidyl choline. This enhanced activity was observed during incubation periods of 1-3 hours and occurred with prolactin concentrations ranging between 2.5 and 100 mug per 300 mul. The enhanced rate of arachidonic acid release from phosphatidyl choline suggests that prolactin stimulates phospholipase A activity and this may be the primary site of action of prolactin in the mammary gland. Specificity of the prolactin effect is suggested by the fact that growth hormone at 100 mug per 300 mul had no effect on the rate of [3H]-arachidonic acid release from phosphatidyl choline.


Testicular LH binding in the hamster: modification by photoperiod and prolactin.

Suppression of testicular weight and activity induced in the hamster by light deprivation can be partially reversed by treatment with prolactin (PRL). The present study investigates the possibility that the stimulatory effect of PRL in this preparation may be mediated through increased LH binding. Hamsters exposed to 5 h light per day for two months to induce gonadal atrophy were injected daily for 2 1/2 weeks with saline, 250 mug PRLP, 20 MUG LH+150 mug FSH, or PRL+LH+FSH. Short light control animals exhibited significantly less LH binding than controls on 14 h of light per day. Treatment with LH+FSH had no effect on LH binding while PRL alone or in combination with LH+FSH increased binding to levels greater than the long light controls. Peripheral testosterone concentrations reflected the level of LH binding. Scatchard analysis indicates that the decreased binding in the short-day animals is due to reduced LH receptor numbers and that PRL treatment elevates receptor levels thereby increasing LH binding. These results suggest that the mechanism by which PRL stimulates testicular function in hamsters with regressed gonads is through increased binding of endogenously produced LH.


Effects of an estrogen antagonist on enzyme activities and [3H]estradiol nuclear binding in uterus, pituitary and brain.

The estrogenic and anti-estrogenic properties of CI-628 on these specific biochemical markers of estrogen action was not identical in the different target tissues. The CI-628 dependent changes in protein content and enzyme activities in the uterus were less than half of the EB changes. In the preoptic area and CM-amygdala, CI-628 resulted in approximately the same changes in CAT and MAO activity as EB. 3) In the pituitary and uterus, CI-628 antagonized the EB-dependent increase in G6PDH activity. In the CM-amygdala and preoptic area, CI-628 pretreatment did not anatagonize the EB dependent changes in CAT and MAO. 4) CI-628 reduced nuclear binding of[3H]estradiol to approximately 5% of control in the uterus and pituitary and this effect lasted for at least 96 h. In contrast, CI-628 reduced nuclear binding in the preoptic area-hypothalamus-amygdala to 13% of control at 18 h after CI-628, but at 96 h nuclear binding of [3H]estradiol had recovered to 70% of control. Thus, the effect of CI-628 ON THESE SPECIFIC BIOCHEMICAL MARKERS OF ESTROGEN ACTION WAS NOT IDENTICAL IN THE DIFFERENT TARGET TISSUES. The possibility is discussed that the basis for the heterogeneity of responses may reside in the different time course of CI-628 interaction with cellular estrogen receptors in the target tissues.or the heterogeneity of responses may reside in the different time course of CI-628 interaction with cellular estrogen receptors in the target tissues.


Liver protein kinase activity and triiodothyronine.

Protein kinase activities were determined in liver from normal, thyroidectomized and triiodothyronine (T3)-treated rats. Changes related to thyroid hormone were observed in cytosol and nuclear protein kinase activities. When protamine was used as substrate for phosphorylation, thyroidectomy induced a decrease of protein kinase activity associated with nuclei but an increase of activity was found in the cytosol. Fifteen hours after injection of T3 the levels in nuclei and cytosol were restored to normal. When casein was used as substrate, hypothyroidism led to a lowering of protein kinase activity in both fractions and T3 treatment augmented the activity in both. These studies suggest that thyroid hormones modify hepatic protein kinase activity. Results differ depending upon the substrate used. The hormones also appear to alter the subcellular distribution of some protein kinase activities.


Appearances of responses to glucagon in cultured neoatal rat heart cells.

Heart cells from neonatal rats have been cultured. The ability of 10(-5)M glucagon to stimulate adenylyl cyclase activity, and to increase the cAMP concentration and the beating rate in these cells was followed as a function of time in culture. The cultured cells show no response to 10(-5)M glucagon until 5 weeks. By contrast, the cells do respond to 10(-5)M epinephrine with an increase in beat rate, adenylyl cyclase activity and cAMP levels when freshly prepared or after 1 week in culture. Previous studies on the newborn rat heart, acutely isolated, have also shown that the neonatal rat heart is insensitive to glucagon until 4-5 weeks after birth. We conclude that the cultured neonatal rat heart cells can also mature in the same time frame with respect to a glucagon response.


Presence of two neurophysins in the human pineal gland.

The tentative identification of two neurophysins in human pineal glands is reported. The presence of these carrier-proteins for neurohormones was demonstrated by two different radioimmunossays: one highly specific for human pituitary estrogen-stimulated neurophysin (h-ESN) and the second for pituitary bovine neurophysin II (b-NII). These neurophysins accounted for about 0.425% of the total soluble pineal proteins. They were eluted from Sephadex G-75 column with the same elution volume as 125I labeled h-ESN. Their dilution curves were parallel to the standard curves of h-ESN and b-NII, respectively. Electrophoresis on polyacrylamide gel separated two distinct neurophysins. The less anodic neurophysin corresponding to b-NII and the faster moving one to h-ESN. The two proteins were named human pineal neurophysins I and II (h-PNI and h-PNII) in the order of their electrophoretic mobility. Purification of pineal neurophysins by isoelectric focusing resulted also in the separation of two neurophysins: one, recognized by antibody against h-ESN with an isoelectric point (pI) of 4.6, the other recognized by antibody against b-NII with a pI of 4.9. The presence of neurohormone(s), assessed by radioimmunoassay for [8-arginine] vasopressin, was also demonstrated. Under optimal conditions for the association the neurophysin and the neurohormone were eluted from Sephadex columns as a complex. This complex could be dissociated in 0.1N formic acid and the neurophysin and the neurohormone separated on Sephadex G-75.


Naphthylamidase (arylaminopeptidase) activities in rat cartilages and bone.

Extracts of epiphyseal articular and costal cartilages and of metaphyseal bone showed different profiles of activity against 21 aminoacyl beta-naphthylamides. Except for the activity against arginyl and lysyl beta-naphthylamides, most of the activities required the presence of Triton X-100 for extraction, suggesting that in situ, the enzymes are bound to or within membranous structures. Histochemical studies demonstrated that most, if not all, activity was intracellular. The activity is enhanced by Co2+. TAME (2 X 10(-4) mol/l) inhibits activity against leucyl beta-naphthylamide and some others, but has little, if any, inhibition of activity against alanyl beta-naphthylamide. Gel filtration of the extract showed a high molecular weight active fraction eluting with proteoglycan, and an active fraction with an approximate molecular weight of about 230,000. Two active fractions are obtained by gradient elution from DEAE-cellulose.


Effects of aluminium, lead and zinc on delta-aminolaevulinic acid dehydratase.

The effects of aluminium, lead and zinc on ALA dehydratase were examined in vitro and in vivo. Aluminium alone added to the erythrocyte enzyme assay caused a highly significant increase in enzyme activity. A similar result was found with zinc alone. When lead and aluminium were added together, the inhibition found with lead alone was depressed in an additive manner by aluminium. Zinc and aluminium together showed an activation that was stoichiometrically additive. Studies in vivo using rats injected with aluminium and lead showed similar results for hepatic ALA dehydratase as those found in vitro.


Citrate uptake and oxidation by fragments of rat ventral prostate.

Citrate oxidation was studied utilizing an in vitro preparation of rat ventral prostate which was very similar, with respect to citrate metabolish, to the intact prostate. The rate of citrate oxidation was very slow in comparison to kidney, although citrate entered prostatic tissue and accumulated intracellularly. Citrate was converted to isocitrate at a rate which resulted in a constant citrate/isocitrate ratio over a 10-fold variation in medium citrate concentration. The prostate oxidized significantly more alpha-ketoglutarate and malate than citrate. These results suggested that limited citrate oxidation could account for the accumulation of high prostatic citrate levels.


Maternal serum hyaluronidase activity in pregnancy.

An unselected series of 291 pregnant women was followed during pregnancy. Maternal serum was assayed for both hyaluraonidase activity and HPL. Urine was assayed for 24 h oestrogen output. A retrospective examination of the results showed that hyaluronidase activity did not provide as good an indication of fetoplacental well-being as did either HPL or pregnancy oestrogen. There were differences, however, in the mean levels obtained in complicated pregnancy, compared to the levels in normal pregnancy. Fetal death was associated with increased hyaluronidase activity whereas fetoplacental dysfunction was associated with lower levels of hyaluronidase activity.


Enzymes of citrate-cleavage pathway in liver of subjects with adult-onset diabetes.

In the liver of adult diabetics, the activity of two enzymes of the citrate-cleavage pathway was increased, the change being statistically significant for NADP-malate dehydrogenase (+ 46%, p less than 0.05) but not significant for ATP citrate-lyase (+ 55%, p greater than 0.10). The increased activity of NADP-malate dehydrogenase, together with the previously described elevation of pentose cycle dehydrogenases, suggests enhanced NADPH generation. Considering the recently proposed possibility of extramitochondrial acetyl-CoA formation by routes other than the citrate-cleavage (i.e., via cytoplasmic acetyl-CoA synthetase), our data is consistent with the occurrence of increased lipogenetic capacity.


A clotting defect in an Arab colt foal.

A multiple clotting defect in a 3 month old Arab colt foal associated with a deficiency in Factors VIII, IX, and XI is described. No abnormalities in clotting factors were detected in the colt's sire, dam, half-sister and half-brother.


A method of ether anaesthesia in ponies.

A method of inhalation anaesthesia using diethyl ether, following induction with thiopentone, is described in ponies. The high concentrations of ether needed to maintain anaesthesia were obtained by using a Marrett head in circle vaporizer. This methods would only be used in ponies, since the diameter of the tubing is too small for use on larger horses.


An experimental approach to surgery of the equine fetlock and its associated structures.

An illustrated approach to the medial aspect of the equine fetlock joint is described, whereby the collateral ligaments are transected. This approach is suggested as a possible technique for metatarsal or metacarpal bone fracture repair by retrograde pinning, arthrodesis of the fetlock joint and other joint and sesamoid bone surgery. The histological evidence of satisfactory healing is presented and the advantages and disadvantages are discussed.


Veterinary aspects of doping.

Doping can improve or impair performance and can be done either deliberately or accidentally. Accidental doping to win is the offence which most concerns the veterinary surgeon. The distinction between legitimate therapy and assisting an unfit horse to win a race by giving it a drug is a fine one. General guidelines are presented for the veterinary surgeon in practice.


Liver fluke infection in horses and ponies.

Thirty eight cases of Fasciola hepatica infection in horses with associated clinical signs are reported. A method of examining large amounts of faeces for fluke is described. A safe method of treatment for infected horses is given which involves oral medication with oxyclozanide at a dose rate of 15 ml/50 kg body weight.


Rupture of the diaphragm in the horse.

Four cases of fatal diaphragmatic rupture in the horse are described. In 2 cases there was no history of injury but the other 2 animals had sustained recent thoracic trauma. Three of the horses had signs of colic and bowel obstruction complicated, in 2 cases, by respiratory embarrassment; the fourth died rapidly, presumably of shock. Previously published cases are reviewed and the causes, clinical effects, diagnosis and treatment of the disorder are discussed.


Ataxia in a Welsh cob filly due to a venous malformation in the thoracic spinal cord.

An 8-month old Welsh Cob filly had shown progressive ataxia of the hindquarters since about 3 months of age. Autopsy revealed a venous malformation resulting in malacia in the ninth thoracic segment of the spinal cord. It is proposed that the tissue destruction associated with this congenital lesion gave rise to the ataxia.


A review of the diagnosis and treatment of fluid and electrolyte disorders in the horse.

Disturbances of fluid and electrolyte balance may occur in the horse as a consequence of many disease processes. Disorders may be classified into 3 main categories, primary water loss, mixed water and electrolyte depletion and acid base disturbances. This paper reviews the changes which occur with each type of disorder and discusses methods of diagnosis and assessment of such imbalances. The principles of treatment are described and the management of fluid replacement therapy in 3 of the disorders, mainly acute intestinal obstruction, inability to drink and diarrohea is discussed.


Thyroid and testicular hormone responses to graded and prolonged exercise in man.

Eight men were studied during graded (47, 77 and 100% of maximal oxygen uptake) and prolonged (76%) exhaustive treadmill running. Plasma catecholamine levels increased progressively with intensity and duration of exercise. Serum concentrations of thyroid-stimulating hormone (TSH) increased with increasing work loads and were 107 (58--243)% (P less than 0.001) above resting values after maximal work. Thyroxine, triiodothyronine and luteinizing hormone in serum never changed significantly. While a small increase in testosterone concentrations (13 [1--24]%) after maximal exercise probably could be explained by changes in plasma volume, a definite increase (31 [14--56]%) occurred after 40 min of prolonged exercise. During continued exercise testosterone concentrations then gradually declined. Testicular stimulation by the increased catecholamine concentrations possible contributed to the rise in testosterone concentrations, but no evidence was found for a direct catecholamine induced increase in the activity of the thyroid gland. The exercise induced increase in TSH levels possibly explians the increased thyroid hormone secretion rate, which previously has been found in individuals participating in physical training programs.


Perceived exertion of absolute work during a military physical training program.

The purpose of this study was to compare the rating of perceived exertion (PRE) and heart rate (HR) in two groups of 60 military personnel who differed in their level of fitness as determined by maximal oxygen uptake (VO2 max). At an initial testing period (T1), Group I represented a sample of personnel not participating in a training program while Group II had engaged in an endurance program (2--4 mile run/day) for 5 months. Six months later (T2), Groups I and II were retested after having participated in the program for 6 and 11 months, respectively. RPE and HR were measured at the end of each min of a 6 min run at an absolute workload of 6 mph, 0% grade on the treadmill. At T1, Group II had a significantly lower HR at each min of work but no difference existed in RPE between groups at any time during the run. At T2, both groups showed a significant decrease in HR and RPE during each min when compared lobgitudinally. The data suggest that the perception of the intensity of absolute work does not differ in groups differing in their level of fitness when studied cross-sectionally. However, significant reductions in perceived exertion occur following physical training.


Submaximal working capacity, heart size and body size in boys 8-18 years.

Heart diameters, heart volume (HV), PWC130, VO2 at 130 heart rate, and cardiorespiratory reactions during work at 3 kgm-s-1 were obtained in 237 boys ranging in age from 8--18 years. Results indicate that heart size, PWC130, VO130, and exercise HR, VO2/HR, f and SBP change significantly  with age. On the other hand, HV - kg-1 and work VO2, VE and VE/VO2 remain rather stable throughout the growth period. Correlation analysis indicates that about 85% of the observed variation in the size of the heart during growth can be accounted for by the body weight, while about 70% of the variation in light submaximal working capacity (VO130) can be explained by HV alone. Holding age, height and body weight constant by partial correlation procedures yeilds significant relationships between HV and VO130 (r = 0.461), and between HV - kg-1 and VO130 (r = 0.414). Age, height, weight and size of the heart correlated simultaneously against VO130 account for 75% of the variance in the dependent variable. It would seem important to suggest the need for study of the interactions between age, size and maturity, in addition to indicators of size and efficiency of the oxygen delivery system, and indices of muscle oxygen utilization efficiency. Such an approach will permit a more definite partitioning of the variance in submaximal aerobic capicity during growth, and would probably yeild a more conservative estimate of the relationship between the size of the heart and submaximal working capacity during growth.


Estimation of body density and lean body weight from body measurements at high altitude.

Body density and other anthropometric data were obtained on 101 Indian soldiers who were continously staying at high altitude (3920 m) for more than 10 months. Use was made of a human body volumeter, and body density was calculated from observed body weight and volume. Measurements were taken on the body using standard techniques. A stepwise linear regression analysis was performed to establish possible relationships of 36 body measurements with density and lean body weight. Thigh anterior, juxta-nipple skin folds and forearm and ankle circumference were selected in the regression equation predicting body density. Multiple correlation coefficient (R) equal to 0.765 was obtained for this equation. For the predicted lean body weight, R equalled 0.930. The regression equations included body weight, thigh, anterior and juxta-nipple skin fold thicknesses, and forearm circumference. Contribution of other body measurements in the regression of these parameters was not significant. The analysis also revealed that a new set of coefficients is required for the measurements included in the published regression equations.


Maximal O2 uptake of boys and girls -- ages 14--17.

White high school girls (n = 120) and boys (n = 120) aged 14--17 years, selected from 9th, 10th, 11th and 12 grades of a northern, midwest U.S. high school performed running exercise on a motor driven treadmill for determinations of maximal O2 uptake (VO2 max). The mean VO max for all age groups was 40.8 +/- 4.0 and 54.7 +/- 5.6 ml/kg-min-1 for girls and boys respectively. The difference in VO2 max across age groups varied only from 40.2--41.2 ml/kg-min-1 for girls and 54.0-56.3 ml/kg-min-1 for boys. These differences were not significant (P greater than 0.05). The reported VO2 max data are compared with those reported in other studies for bicycle ergometer and treadmill excercise using similar age groups.


[On tremor during movement (author's transl)].

An analysis of the movement of stretching the right forearm showed that the frequency of tremor slightly decreases, whereas the average (absolute) acceleration of tremor and the average electromyoactivity (EMA) of the agonist increase as expected.


Oxygen transport system during exhaustive exercise in Japanese boys.

The study was designed to investigate the age-related increase in maximal oxygen uptake with special reference to the other physiological parameters (ventilation, pulmonary diffusing capacity, cardiac output etc.). 77 normal boys aged 10-18 years were tested by the progressive loading method on a bicycle ergometer. VO2 max increased by approximately 55% with the age from 10-18 years. The increase in VO2 max was accompanied with increase in ventilation (49%), pulmonary diffusing capacity (45%) and cardiac output (5%).


Role of thyroid gland on the peroxidase and iodinating enzymes of submaxillary gland.

The peroxidase (EC 1.11.1.7) and iodinase (EC 1.11.1.8) activities of rat submaxillary gland were found to be increased after thyroidectomy. The enzyme activities were maximal on the seventh day after operation and then decreased slightly. However, the enzyme activities were still more than 100% even 28 days following operation. Administration of thyroxine (10mug/100 g body weight) prevented the increase. Puromycin, cycloheximide, and actinomycin D, the inhibitors of protein synthesis, as well as thiouracil partially abolished the increase of activities. These results suggest that thyroxine acts as a regulator of the iodinase and peroxidase enzyme(s) of submaxillary gland,


Effect of 2-deoxy-D-glucose on the cell-surface glycoproteins of hamster fibroblasts,.

1. Growth of baby hamster kidney (BHK) cells in medium containing 2-deoxy-D-glucose is retarded in direct proportion of the 2-deoxyglucose concentration. The severity of the effect is reduced in medium containing high relative concentrations of glucose. 2. 2-Deoxyglucose inhibits the incorporation of radioactivity from mannose, galactose, glucosamine, fucose and N-acetylmannosamine precursors into acid-insoluble cellular material. Incorporation of radioactively labelled leucine into protein is not affected by 2-deoxyglucose. 3. BHK cells grown in the presence of 2-deoxyglucose become less sensitive to the toxic action of certain plant lectins, ricin of Ricinus communis and Phaseolus vulgaris phytohaemagglutinin, which bind specifically to cell surface galactose and N-acetyl-galactosamine residues. By contrast, 2-deoxyglucose increased the sensitivity of BHK cells to the weak toxicity of concanavalin A, which binds to surface mannosides. Treated cells also become more agglutinable with concanavalin A. 4. Cell surface glycoprotein labelled by lactoperoxidase-catalysed iodination have been examined by dodecylsulphate-polyacrylamide gel electrophoresis. The radio-iodinated glycoprotein prepared from cells grown in medium containing 2-deoxyglucose migrate more rapidly than glycoproteins from cells grown in the absence of inhibitor.


Properties of a baby-hamster-kidney cell line with increased resistance of 2-deoxy-D-glucose.

A cultured cell line with increased resistance to 2-deoxy-D-glucose was obtained from cloned baby hamster kidney fibroblasts, BHK 21/C13, after repeated exposure to high concentrations of 2-deoxyglucose. The increased resistance could not be attributed to a decreased permeability of deoxysugar. The resistant cell line incorporated radioactive 2-deoxy-Dglucose in glycoproteins at a similar rate as parental BHK 21C13 cells. Incorporation of radioactive glucosamine, galactose and to lesser extent mannose into cellular glycoproteins was inhibited by 2-deoxyglucose to similar extents in the resistant cells and parental BHK 21/C13 cells. Changes induct cells were detected by altered to toxic plant lectins and by surface labelling as described for parental cells in the preceding paper. It is suggested that the toxicity of 2-deoxy-D-glucose to normal fibroblasts is not mediated through effects on glycosylation of cellular glycoproteins.


On the role of ferredoxin and ferredoxin-NADP+ reductase in cyclic electron transport of spinach chloroplasts.

Antibodies prepared against purified spinach ferredoxin and ferredoxin-NADP+ reductase were used as specific inhibitors of electron-transfer reactions dependent on either ferredoxin or ferredoxin-NADP+ reductase; The possible role of both electron carriers in cyclic electron transport was checked using cytochrome b6 photoreactions as indicator. It could be demonstrated that the ferredoxin antibody inhibits cytochrome b6 photoreduction. Ferredoxin-NADP+ reductase, however, appears not to be involved in this pathway: reductase antibody in concentrations sufficient to completely inhibit electron transport to NADP+ had no effect on cytochrome b6 photoreduction. Quantitative treatment of the immunoassay data showed that osmotically shocked chloroplasts contain both bound ferredoxin and ferredoxin-NADP+ reductase in concentration approximately equal to that of cytochrome b6.


RNA polysomes and RNA synthesis in the silk glands of the silkworm Bombyx mori;.

Total RNA polymerase activity, as well as the proportion of alpha-amantin-sensitive and resistant during activity, have been measured in the posterior silk glands of the silkworm as a function of growth the fifth larval instar. During the first 5 days, termed the growth phase, the total enzyme activity and particularly the portion that is alpha-amantin-resistant increases to reach a peak value and thereafter declines during the secretory phase, Much of the enzyme remains firmly bound and insoluble. Heparin only only does not inhibit this insoluble and probably chromatin-bound activity which would indicate lack of initiation, but it enhances the activity. A large proportion of newly transcribed RNA is released from the transcription complex. The synthesis of RNA has been studied both qualitatively and quantitatively during the same period. RNA synthesis becomes important on the second day of the fifth instar, as does the RNA polymerase, and stays at a high level for several more days. The results from these studied as well as those with incorporation of 32P indicate interference of varying precursor pools in quantitatively measured RNA synthesis. However, RNA content as well as RNA synthesis in vitro show a close correlation with RNA polymerase activity. The labeled RNAs extracted at different days of the fifth instar have been fractioned on sucrose gradients; this demonstrated that the predominant product of RNA synthesis, as followed by [3H]uridine incorporation at short time intervals, is 45-S preribosomal RNA and 4-5 S RNA. The 45-S RNA is transformed to 19-S and 30-S ribosomal RNA as time progresses or after a chase with unlabeled and/or actinomycin D. There also exists a component heavier than 45S which is fairly rapidly labeled to a small extent.


Fate of histone messenger RNA in synchronized HeLa cells in the absence of initiation of protein synthesis.

The fate of cytoplasmic histone mRNA was studied under conditions in which initiation of protein synthesis in synchronized HeLa cells is S phase was blocked by increasing the osmolarity of the growth medium with NaCl. In contrast to the interruption of DNA replication with hydroxyurea, which results in an exponential degradation of translatable histone mRNA with a half-life of about 10-13 min, blocking the initiation of protein synthesis leads to only a marginal loss of biologically active histone mRNA in the cytoplasm. When the initiation of protein synthesis was interrupted by treating cells with 150 mM NaCl, 40-50% of the total cytoplasmic histone mRNA previously translated in polyribosomes appears in the cytoplasm integrated into mRNA-protein particle(s) sedimenting between 15 S and 30 S. On the other hand, in untreated S-phase cells or in cells blocked with hydroxyurea only 3-6% of the total translatable histone mRNA is found in the cytoplasm not bound to ribosomes or their subunits. In addition, the degradation of histone mRNA in hydroxyurea-blocked S-phase cells is prevented when the initiation of protein synthesis is inhibited with NaCl. These studies clearly indicate that the inhibition of initiation of protein synthesis per se is not the cause for the rapid degradation of cytoplasmic histone mRNA observed when DNA replication is turned off and that the inactivation of these mRNAs is a process dependent on continuous protein synthesis.


Progesterone receptors in the chick oviduct. Determination of the total concentration of binding sites in the cytosol and nuclear fraction and effect of progesterone on their distribution.

1. Exchange techniques were developed for measurement of total progesterone receptor binding sites concentration in the cytosol and nuclei of chicken oviduct. 2. The level of progesterone receptor in the cytosol was under both oestrogen and progesterone control. Primary stimulation by oestradiol benzoate increased the receptor concentration from approximately 10 000 sites/cell to approximately 40 000 sites/cell in the magnum cytosol; subsequent withdrawal from oestrogen treatment led to a decrease to approximately 14 000 sites/cell after 6 weeks. After progesterone administration (3 mg/kg) to oestrogen-stimulated, withdrawn chicken, the receptor concentration decreased to approximately 40% of the initial level within the first 4 h; afterwards the receptor level rose again and by 40 h exceeded slightly the initial one. 3. The nuclear levels of the receptor reached a maximum at 1 h after the progesterone injection; however, the gain of the binding sites by the nuclei (approximately 900/nucleus) did not account for their loss from the cytosol. The maximum nuclear receptor level was not influenced by the dose of progesterone within range of 1-10 mg/kg. 4. The extractibility of the nuclear progesterone receptor by 0.5 M NaCl was strongly influenced by preincubation of the nuclei at 30 degrees C; irrespective of this preincubation, approximately 12% of the nuclear receptor resisted extraction with 2 M NaCl/5 M urea.


Nucleotide sequence (n=159) of the amino-acid-accepting 3'-OH extremity of turnip-yellow-mosaic-virus RNA and the last portion of its coat-protein cistron.

The experiments described in this paper and the following one establish the sequence of the 3'-OH terminal 159 nucleotides of turnip yellow mosaic virus RNA. Uniformly 32P-labeled turnip yellow mosaic virus RNA was partially digested with T1 ribonuclease and the fragments were fractionated by polyacrylamide gel electrophoresis. Fragments originating from the 3'-OH end of the RNA molecule were identified by testing for the 3'-terminal oligonucleotide, C-COH, after total U2 ribonuclease hydrolysis. Once identified, the 3'-OH terminal fragments were sequenced by the methods of Sanger et al. The first 51 nucleotides of the longest of the sequenced fragments (158 nucleotides) extends into the 3'-terminal part of the coat protein cistron. The coat protein cistron is followed by a stretch of 108 untranslated nucleotides whose function, though still unknown, is probably linked to the tRNA-like properties which have been attributed to the 3'-OH extremity of this viral RNA. Two possible secondary structures are proposed for the sequence and the implications of the findings with regard to the tRNA-like properties of the extremity are discussed.


Subcellular distribution of aminoacyl-tRNA synthetases in various eukaryotic cells.

The total amount, size distribution and binding of aminoacyl-tRNA synthetases to ribosomes in a variety of mammalian and avian cells was studied under standard conditions of sample preparation and assay. Aminoacyl-tRNA synthetases appear to exist in three general forms; 'free' enzyme of about 4-9 S, one or more 'enzyme complexes' of about 18-25 S, and in association with ribosomes. The aminoacyl-tRNA synthetase activity for many individual amino acids was surprisingly similar in cell types chosen to be diverse with respect to differentiation state, transformation, and growth rate. Total activity for all amino acids varied about 4-fold, based on a constant volume of cells. Embryonic tissues had a comparatively high proportion of total synthetase activity associated with ribosomes, whereas this value was relatively low for mouse liver. Distinctive distribution patterns with common and variable features were observed for individual enzymes. The only aminoacyl-tRNA synthetases found not to be associated in significant amounts with either 18-25 S enzyme complexes or ribosomes in any of the cell types examined were the enzymes for alanine, histidine, and serine. All cell types evidenced 18-25-S synthetase activity for arginine, aspartic acid, glutamine, glutamic acid, isoleucine, leucine, lysine, methionine, proline, and valine, although in quite variable porportions of the total activity observed for these amino acids. For example, of the valyl-tRNA synthetase activity not associated with ribosomes, 35% and 100% were found to sediment at 18-25 S in Friend leukemia cells and mouse liver respectively. All cells had two easily distinguishable peaks of arginyl tRNA synthetase activity at 4-9S and 18-25S respectively; however, the relative proportion of enzyme activity in the peaks differed between cell types. Phenylalanyl-tRNA synthetase was not observed to occur in an 18-25-S complex in any of the cell types examined but was bound to ribosomes in variable but generally relatively high proportions. Numerous other specific differences are described. No underlying physiological or biochemical principle has been recognized to account for the specific distribution patterns observed. However, they may reflect variations in cellular architecture that may be related to regulation of protein synthesis.


DNA synthesis in HeLa cells and isolated nuclei after treatment with an inhibitor of spermidine synthesis, methyl glyoxal bis(guanylhydrazone).

Addition of methyl glyoxal bis(guanylhydrazone) to HeLa S3 suspension cultures resulted in increased putrescine levels and decreased spermidine and spermine levels preceding a drop in incorporation of [3H]thymidine, [3H]uridine and [14C]leucine into macromolecules. When putrescine, spermidine, spermine or cadaverine was added simultaneously with methyl glyoxal bis(guanylhydrazone), the drug had no detectable effect on the synthesis of macromolecules. In nuclei isolated from cells treated with methyl glyoxal bis(guanylhydrazone) the reduction in the rate of DNA synthesis was equal to the reduction of [3H]thymidine incorporation in the corresponding whole cells. The capability of the nuclei to synthesize DNA could not be restored by adding spermidine or spermine to the system in vitro. The rate of DNA chain elongation was only reduced slightly by methyl glyoxal bis(guanylhydrazone) indicating that decreased levels of spermidine and spermine lead to a decrease in the number of replication units active in DNA synthesis within each cell.


Characterization and partial purification of an inducible protein related to hexose proton cotransport of Chlorella vulgaris.

1. Cells of Chlorella vulgaris induced for hexose transport contain a membrane-bound protein component that is missing in non-induced cells. This was shown by double labelling experiments with [14C]phenylalanine and [3H]phenylalanine applying the method of Kolber and Stein [Nature (Lond) 209 (1966) 691]. The specific protein is completely absent from soluble fractions. 2. An enrichment of the double-labelled peak was observed in membrane fractions enriched in plasmalemma. The best purification was obtained when cell walls were purified; the residual membranes attached to the walls contained the transport protein with a 12-times-higher specific activity than the crude extract. 3. The transport protein had the characteristics of an intrinsic membrane protein. Its molecular weight was found to be 30 000 on sodium dodecylsulphate gels. The protein did not show sugar binding activity, however. 4. Induced cells lose their state of induction with a half life of about 4 h; 7 h after induction the double-labelled transport protein is no longer detectable.


Accumulation of tRNAMetf on 80-S ribosomes in vitro under the influence of a Met-tRNA deacylase from rat-liver microsomes.

A Met-tRNA deacylase has been partially purified from the 0.5 M KCl wash of rat liver microsomes. In preparative sucrose gradients, the active component sediments as a single band at about 6 S, corresponding to an estimated molecular weight of 1.7 X 10(5). The deacylase is specific for Met-tRNA, without discriminating between Met-tRNA f and Met-tRNAm. Met-tRNAf bound to the initiation factor IF-MP in the ternary complex, IF-MP-GTP-Met-tRNAf, or to initiation-factor-dependent, complexes with 40-S subunits or 80-S ribosomes, is protected against deacylation. However, in the course of the initiation-factor-dependent joining of the 40-S subunit complex to 60-S ribosomal subunits, the bound Met-tRNAf is exposed to added deacylase. Under these conditions, deacylation is inhibited by GTP. The tRNAMetf remains bound and accumulates on the 80-S ribosomes.


Forms AI and AII DNA-dependent RNA polymerases as components of two defined pools of polymerase activity in mammalian cells.

Two species of form A (or I) RNA polymerase have been identified in eucaryotic cells and there is evidence that this alpha-amanitin-insensitive activity exists as two discrete pools: a pool of 'free' activity, which is identified by its ability to transcribe poly d(A-T) in the presence of actinomycin in vitro, and a pool of enzyme in the form of a transcription complex ('engaged') which is unaffected by inhibitors of initiation of RNA synthesis. 1. The principles underlying and the practical application of the technique used to define the pool of 'free' RNA polymerase activity have been analysed in considerable detail. On the basis of actinomycin titrations of poly[d(A-T)]dependent activity in isolated organelles, it is concluded that a pool of 'free' RNA polymerase A activity exists in mammalian nuclei which, under certain circumstances, is lost from nuclei during their isolation. The evidence presented suggests that nucleoli, resolved from nuclei by the classical sonication technique, contain form A polymerase exclusively in the transcription complex form. 2. Different techniques used to solubilise RNA polymerase activity from nucleoli are shown to give rise to different proportions of the two form A RNA polymerase species (AI and AII, as defined by their differential elution from phosphocellulose): whereas low-ionic-strength extraction gives rise to form AII, high-salt, sonication extracts contain predominantly the form AI enzyme. It is shown that the sonication technique results in the conversion of form AII to form AI. By a careful appraisal of the products of these procedures and a novel polymerase solubilisation technique, it is concluded that RNA polymerase AII is the 'engaged' form of the enzyme found in the transcription complex. 3. Making use of the finding that the 'free' form of the enzyme is lost to the cytoplasmic fraction on nuclear isolation, this activity has been characterised without the requirement for solubilisation techniques which might result in the conversion of one form to another: the 'free' species is shown to be form AI RNA polymerase. 4. These conclusions that two discrete pools of form A RNA polymerase activity contain different species of the enzyme are briefly discussed in the light of other published information concerning their subunit structures and their potential role in the expression of the ribosomal RNA coding sequences.


On the involvement of cytochrome P-450 in the binding of ribosomes to a subfraction of rat-liver rapidly sedimenting endoplasmic reticulum.

Rat liver endoplasmic reticulum has been separated into four ribosome-containing subfractions, two from rapidly sedimentation endoplasmic reticulum and two from the microsomes, by differential centrifugation and sucrose density centrifugation. Ribosomes from one of the rapidly sedimenting subfractions were extracted by Trion X-100 as a complex with cytochrome P-450, optimally at a detergent protein ratio of 2/1 (w/w). Upon extraction approximately 50% of the cytochrome P-450 in the membrane appeared complex-bound to ribosomes, and, maximally, 6-7 subunit molecules of the cytochrome were attached per ribosome. The specific concentration of cytochrome P-450 on these ribosomes was 2.5-times higher than in the parent membrane. Cytochrome b5, glucose-6-phosphatase, NADPH-cytochrome c reductase, NADH-ferricyanide reductase, cytochrome oxidase and phospholipids were present in small or trace amounts on the ribosomes in relation to cytochrome P-450. Ribosomes extracted from other subfractions contained much less bound cytochrome P-450. Phenobarbital treatment induced an increase in the cytochrome P-450 content that was different for the various subfractions. This increase could not be correlated with changes in the amounts of cytochrome-ribosome complexes released by detergent. We propose that cytochrome P-450 is part of a specific binding site in the membrane for a fraction of the ribosomes attached to the endoplasmic reticulum. The ribosomes may be anchored to cytochrome P-450 via nascent chain proteins.


Inversion of the unnatural cis C/D sterol ring junction of 5alpha, 14beta-cholest-7en-3beta-ol by rat-liver enzymes.

Labelled 5alpha, 14beta-cholest-7-en-3beta-ol with the 'unnatural' cis C/D ring junction was synthesized. Incubations of this sterol with rat liver homogenate under aerobic conditions gave radioactive 5alpha-cholest-7-en-3beta-ol, 5alpha-cholest-8(14)-en-3beta-ol andcholesterol indicating the presence of enzymes in the rat liver capable of inverting the C/D cis to a trans configuration. No radioactivity was found associated with added 14beta-cholesterol showing the specificity of the 14alpha configuration for the enzymic conversion of a sterol molecule into cholesterol.


Structural studies of yeast flavocytochrome b2: cooperative roles of the alpha and beta globules in the formation of the flavin-binding sites.

The purpose of the study reported here was the localization of the heme binding sites on the two globular fragments, alpha and beta, of the 'cleaved' form of the flavocytochrome b2 chain. These fragments were partially resolved by means of molecular sieving under denaturing conditions (3 M or 6 M guanidine in the presence of 2-mercaptoethanol). They were then renatured in the presence of excesses of FMN and protoheme. The protoheme was found to be quantitatively bound to the alpha subunit, confirming previous findings. The flavin binds neither to alpha alone nor to beta alone, but only to the reassociated alphabeta protomer. the results are discussed in terms of the possible occurrence of gene fusion in the formation of the complex flavocytochrome chain of this very particular L-lactate cytochrome c reductase found specifically in yeasts.


alpha-Mannosidase from Phaseolus vulgaris. Composition and structural properties.

Both alpha-mannosidases I and II from Phaseolus vulgaris have molecular weights about 210000-220000 and contain approximately 2 mol zinc/mol protein. alpha-Mannosidase I seems to consist of more glutamic acid than alpha-mannosidase II, while the latter is richer in serine. They are glycoproteins: alpha-mannosidase I contains 8.3% carbohydrate by weight while alpha-mannosidase II contains 16.5%. This enzyme form shows a greater thermal stability than alpha-mannosidase I. The structure of alpha-mannosidase has been investigated by equilibrium sedimentation analysis in guanidine hydrochloride, electrophoresis in dodecylsulphate, and alkaline electrophoresis after exposure to high pH. The protein appears to be composed of two non-covalently bound subunits of molecular weights about 110000. Electron micrographs revealed images of molecules that consisted of two rod-shaped monomers of roughly square cross-sections 4.2 X 4.2 nm. Each rod was about 7.4 nm long. The monomers seemed parallell along the long axis.


Monitoring of the specific radioactivity of S-adenosylmethionine in kidney in vivo.

The specific radioactivity of S-adenosylmethionine was followed in the cat kidney during the infusion of L-[Me-3H]methionine into the corresponding renal artery. For this purpose 14C-labelled 4-(2-aminoethyl)pyrocatechol([14C]dopamine) as methyl acceptor was injected locally every 15 min and the 3H and 14C activity of the methylation product homovanillic acid, isolated from urine, was measured. Approximately 5% of the 14C label is excreted during the first renal passage as [14C]homovanillic acid. The specific activity of S-adenosy[Me-3H]methionine in the kidney was calculated from the known specific radioactivity of [14C]dopamine injected and the measured radioactivity ratio, 3H: 14C, of homovanillic acid isolated from urine. The specific activity of S-adenosyl[Me-3H]methionine reaches a constant value in kidney about 30-60 min after the beginning of the L-[Me-3H]methionine infusion. This plateau value was 28% +/- 14% (n = 5) lower than the specific activity of L-[Me-3H]methionine in the venous blood from the corresponding kidney. The difference between the specific radioactivity of S-adenosyl[Me-3H]methionine in kidney and of free methionine in plasma is explained by the existence of a methionine source of minor specific activity in the kidney. The average life span of S-adenosylmethionine in the kidney is 19.5 +/- 8.7 min (n = 5).


A single-strand-specific DNA-binding protein from mouse cells that stimulates DNA polymerase.

A protein of 30000-35000 molecular weight was isolated from mouse ascites cells. This protein binds preferentially to single-stranded DNA. Evidence is presented that this protein maintains single-stranded DNA in an extended configuration. In the presence of single-stranded template the protein stimulates mammalian DNA polymerase alpha but not the mammalian DNA polymerase beta and not some microbial DNA polymerases. The protein is phosphorylated in vitro by a chromatin-associated protein kinase. The modified DNA-binding protein does not stimulate the DNA polymerase alpha.


Electron microscopy of negatively stained jackbean urease at three levels of quaternary structure, and comparison with hydrodynamic studies.

Electron microscopy, with sodium phosphotungstate as negative stain, has been carried out on purified jackbean urease prepared at three levels of quaternary structure: (a) A1 urease, Mr = 240 000, S20,W = 11.5 S (b) alpha urease, Mr = 480 000, S20,w = 18.3 S (c) polymers of alpha urease above the tetramer stage. The compatibility of the images from level to level leaves no doubt that the enzyme itself is being visualized, and the following geometry is suggested by electron microscopy: A1 molecules are cyclic trimers, which pair up in eclipsed position across a 1-nm cleft to form the hexameric alpha, which displays D3 (or 32) symmetry of a trigonal prism. Polymers consist of alpha molecules aligned with their clefts coplanar and an angle of 120 degrees between each triplet of 3-fold axes. These features correspond reasonably well with sedimentation and electrophoretic studies of the solvated enzyme, which have indicated a hemispherical A1, a spherical alpha, and string-of-beads polymers. Sedimentation constants of the urease polymers up through the pentamer level were found to be compatible with the rosette, straight-chain, and zig-zag forms seen in the electron microscope, and with the suggested protomer arrangement in A1 and alpha urease.


Covalent structure of collagen: isolation of chymotryptic peptides and amino acid sequence of the amino-terminal region of alpha2-CB3 from chick skin.

Five chymotryptic peptides were isolated from alpha2-CB3 of chick skin collagen by a combination of ion-exchange and molecular sieve chromatography. Together, they account for the entire amino acid content of the parent peptide. Their alignment was deduced by isolation and sequence analyses of overlapping tryptic peptides. In addition, the amino acid sequence of the three consecutive chymotryptic peptides, containing 132 amino acid residues, from the NH2-terminus of alpha2-CB3 was determined by automated Edman degradation of the peptides and tryptic peptides derived from them. The resulting sequence shows an identity of approximately 80% when compared with those of the homologous portions of rat and calf skin collagen. In contrast, extensive substitutions are present when the sequence was compared with that of the corresponding segment of the alpha1 chain from the same species.


Chemical and physical properties of mycobacteriophage D29.

Mycobacteriophage D29 has a head of uniform size (average diameter 65 nm) and regular shape and a tail of variable length. The stability of the bacteriophage is optimal between pH 9 and 10. The virus contain double-stranded DNA and six structural polypeptides, three major and three minor. The molecular weights of these six polypeptides, as determined by polyacrylamide gel electrophoresis in the presence of dodecylsulfate, are 150 000, 138 000, 13 000, 66 000 and 24 000. The virus contains no lipids as shown by (a) the lack of structural changes after inactivation of the bacteriophage with chloroform, (b) the absence of lipids containing [32P]phosphate or [35S]sulfate in labeled virus, and (c) the absence of an electron paramagnetic resonance spectrum in bacteriophage which had been incubated with a nitroxide-containing fatty acid.


Peptidyl transferase center of rat-liver ribosome cores.

Protein-deficient particles have been obtained by treating rat liver 80-S ribosomes or their 60-S subunits with 1 M NH4Cl in the presence of 50% ethanol at 0 degrees C (Po-cores) and 37 degrees C (P37-cores). The Po-cores from 80-S ribosomes are totally inactive in polyphenylalanine synthesis but fully active in the 'fragment assay' to test peptidyl transferase activity. The polymerizing activity of the cores is restored up to 40--50% of control activity by incubation in the presence of the split proteins. Three proteins are totally lost in the treatment, namely proteins L12, L40/41 ans S25. A series of up to nine different spots in the region of the L40/41 proteins are detected when the split fraction is analyzed by two-dimensional electrophoresis. This series of spots is, however, reduced to only two proteins when the second dimension is carried out in the presence of sodium dodecylsulphate. 80-S ribosome-derived P37-cores are about 80% active in the 'fragment reaction' while 60-S-subunit-derived particles are inactive in this assay. The inhibitory effect of a number of antibiotics is differentially affected by the treatment suggesting different localization of their binding sites. A comparative study of the proteins released by treatment in the two types of particles suggests the involvement in the peptidyl transferase center of the ribosome of one or more of the floolwing proteins: L21, L24, L27, L28 and L36.


Synthesis and turnover of phenylalanine ammonia-lyase in root tissue of sweet potatoe injured by cutting.

1. Antibody toward phenylalanine ammonia-lyase of root tissue of sweet potato injured by cutting was obtained by immunizing of a rabbit with purified enzyme. 2. Phenylalanine ammonia-lyase activity developed markedly in disks of sweet potato roots in response to cut injury, reached a maximum after 12 h, and then decreased thereafter. Phenylalanine ammonia-lyase content, which was determined by the quantitative immunoprecipitation method, changed in parallel with enzyme activity. The rate of incorporation of the label from [14C]leucine into phenylalanine ammonia-lyase initially increased with the time of incubation after slicing, reached a maximum in 6h of incubation, then remained at an almost constant level regardless of the decrease in enzyme activity. The results indicate that the increase in enzyme activity is due to de novo synthesis of phenylalanine ammonia-lyase and the subsequent decrease in activity is not based on decreased synthesis of phenylalanine ammonia-lyase.


The synthesis of several 8-substituted derivatives of adenosine 5'-monophosphate to study the effect of the nature of the spacer arm in affinity chromatography.

Reaction of adenosine 5'-monophosphate (Ado-5'-P) with bromine at pH 4.0 yielded 8-bromo-adenosine 5'-monophosphate and following reaction with several alpha,omega-diaminoalkanes gave the corresponding 8-(omega-aminoalkyl)-Ado-5'-P derivatives. Condensation of these analogues with N-trifluoroacetyl-glycine or beta-alanine in the presence of a water-soluble carbodiimide generated several 8-substituted derivatives. These analogues: (see article) comprised an 8-substituted Ado-5'-P ligand to which a spacer molecule of similar length but differing hydrophobicity was attached. The derivatives were purified, characterised and attached to CNBr-activated Sepharose. The chromatographic behaviour of the resulting absorbents was investigated in terms of their ability to bind both lactate and alanine dehydrogenase. The enzymes bound tighter to the more hydrophobic derivatives with the strength of the interaction decreasing with increasing hydropholicity. The effect of the introduction of a single hydroxyl group in the spacer arm was also studied with several enzymes known to exhibit anomalous behavior on affinity chromatography. In free solution, (Ado-5'-P)-8-NH CH2CH2CH2CH2CH2CH2NH2 and (Ado-5'-P)-8-NH CH2-CH CH2NH CO CH2NH2 were competitive with NADH for rabbit muscle lactate dehydrogenase, (see article) with Ki values of 2.0 and 1.8 mM respectively. These data suggest a difference in accessibility of the hydrophobic and hydrophilic derivatives when attached to Sepharose rather than a fundamental difference in affinity. This suggestion is supported by the fact that there is no obvious correlation between the Ki values in free solution of a series of 8-(omega-aminoalkyl)-Ado-5'-P derivatives of increasing chain length, and hence hydrophobicity, and their chromatographic behaviour when immobilised. The data suggest that the hydrophobicity/hydrophilicity of the spacer arm determined the accessibility of the immobilised ligand to interaction with the complementary enzyme.


Study of protein synthesis in rat liver mitochondria use of cycloheximide.

1. The effects of short-term and long-term administration of cyclohexidine on rat liver mitochondrial protein synthesis have been examined and were found to be different. 2. Long-term administration of cycloheximide resulted in inhibition of total cellular protein synthesis including that of mitochondria while, at short-term intervals, 8-10% of mitochondrial protein synthesis was cycloheximide-resistant. 3. The inhibitory effect was also reflected in terms of protein synthesizing ability of mitochondria in vitro, the inhibition becoming apparent at 40 min and showing progressive increase with time. 4. The observed inhibition of mitochondrial protein synthesis by cycloheximide was not due to either inhibition of energy metabolism or alteration of amino-acid pool. 5. Cycloheximide did not enter mitochondria or sonic preparation under conditions in vitro. On the other hand, after administration of [3H]cycloheximide, significant quantities of the label were found to be associated with mitochondria and mitoribosomes. 6. These results indicated that cycloheximide reached the site of action in mitochondria under conditions in vivo but was unable to do so in vitro. 7. The results are discussed to elucidate the possible mechanisms involved in the inhibition of truly mitochondrial protein synthesis by cyclohexamide.


Entry of mRNA into polyribosomes during recovery from starvation in mouse sarcoma 180 cells.

Starvation in mouse sarcoma 180 cells leads to a block in initiation of protein synthesis which results in the conversion of polysomes to 80-S ribosomes. The mRNA is released as nucleoprotein particles. Addition of nutrients to the starved cell causes the rapid uptake of the mRNA into large polysomal structures. The ribosomes enter these structures more gradually, without much effect on the sedimentation profile of the labeled mRNA. The results indicate that the stored mRNA in starved cells is readily available for uptake into polysomes. They also suggest that the mRNA may be binding to components other than ribosomes during the initial stages of polysome formation.


Structural studies on 2-acetamido-1-N-(4-L-aspartyl)-2-deoxy-beta-D-glucopyranosylamine and 2-acetamido-6-O-(alpha-L-fucopyranosyl)-1-N-(4-L-aspartyl)-2-deoxy-beta-D-glucopyranosylamine by 360-MHz proton-magnetic-resonance spectroscopy.

The 360-MHz proton magnetic resonance spectra of 2-acetamido-1-N-(4-L-aspartyl)-2-deoxy-beta-D-glucopyranosylamine (GlcNAcbeta1 leads to Asn) and 2-acetamido-6-O-(alpha-L-fucopyranosyl)-1-N-(4-L-aspartyl)-2-deoxy-beta-D-glucopyranosylamine (Fucalpha1 leads to 6GlcNAcbeta1 leads to Asn) in deuterium oxide were completely interpreted. The chemical shifts and coupling constants were refined by simulation of the spectra. By means of an adapted Karplus equation the pyranose ring conformation of the sugars was calculated. The change of the geminal coupling constant J6a,6b in the N-acetylglucosamine residue of Fucalpha1 leads to 6GlcNAcbeta1 leads to Asn with respect to GlcNAcbeta1 leads to Asn is proposed to be characteristic for the (1 leads to 6) glycosidic linkage.


Hemihypotrophy in a girl with a translocation t(13q;7p).

A 10 year old girl with a mental age of 7-8 years, normal height and head circumference and several minor anomalies had hemiasymmetry of the entire body, the left side being uniformly smaller than the right. The smaller side was considered the abnormal side and her condition interpreted as hemihypotrophy on the basis of a chromosome abnormality which involved mosaicism, with lymphocytes showing a balanced but very unequal translocation of most of 13q transferred to 7p and both translocation chromosomes being present, and all examined fibroblasts lacking the small translocation chromosome and hence being monosomic for 13p, proximal part of 13q and a terminal portion of 7p.


Fatal CNS dysgenesis with severe microencephaly, mental retardation, seizures and paucity of myelin, autosomal recessive trait?

Siblings are reported with severe mental retardation, spastic cerebral palsy and seizures; in addition they had progressive or intermittent jaundice and recurrent infections; they died at 3 and 4 years respectively. Neuropathological studies in one showed a small brain with an almost complete lack of myelin in cerebral white matter, brain stem, cerebellum and anterolateral parts of the spinal cord. The condition most likely represents a dysgenesis of myelin (dysmyelination), possibly due to an inability of oligodendrocytes to form myelin and/or metabolic defects in the process of myelination. This mental retardation condition is probably inherited as an autosomal recessive trait and may represent a special type of a primary CNS developmental defect.


HLA frequencies in primary immunodeficiency diseases (pidd).

Thirteen patients with primary immunodeficiency disorders and their twenty-five healthy first-degree relatives were tissue typed and their HLA make-up was compared with that of a normal control population. HLA-A2 occured in 92.3% of patients as opposed to 60.8% in the control group (P less than 0.02), HLA-A9 in 7.6% vs. 25% (P less than 0.02) and HLA-B8 in 0% vs. 21% (P less than 0.04). One of the patients with severe combined immunodeficiency showed one "extraneous" HLA specifity.