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.


Digoxin elimination by exchange transfusion. The report covers four cases presenting simultaneous indications for digitalisation and exchange transfusions. Intravenous administration of digoxin was followed: 1. by monitoring of the behaviour of the plasma digoxin level; 2. by determination of the total amount of glycoside eliminated by the blood exchange. Particular attention was paid to the effect of the delay between injection and exchange transfusion on the amount of digoxin eliminated. All four cases showed moderate falls in plasma levels. The amounts of digoxin eliminated by exchange transfusion were in reverse relationship to the delay between administration of digoxin and the blood exchange. At no time did the eliminated fraction exceed 5% of the total amount present in the body.


Nonspecific immunotherapy with BCG vaccine in bladder tumors: a preliminary report. The therapeutic and prophylactic value of nonspecific immunological stimulation with BCG vaccine against papillary bladder carcinomas has been evaluated. Nine patients were treated and followed for periods of 4-9 months. Attempts have been made to compare the effects of BCG vaccine administered via dermal scarifications, or by means of direct intralesional injection into the tumor and/or its base. The initial results obtained so far show the advantages of the intralesional injection method, with perceptible regressions observed in 2 cases. Nonetheless, it is not considered an adequate procedure, since repeated cystoscopies are necessary, and it is not devoid of risks. One patient suffered a severe hypersensitivity reaction. Dermal administration proved a failure in 3 cases where it was used with curative purpose. Sufficient time has not elapsed to allow us to estimate its full prophylactic value in preventing recurrences, although as yet, none of the 4 patients who have been stimulated have shown any signs of recurrence. More time and more cases are required before one can draw valid conclusions, but we consider that dermal scarifications may be useful to patients who have undergone surgery, helping their immunological system to destroy small remnants of malignant cells in the bladder wall, the regional lymph nodes or distant structures, as well as to prevent recurrences in patients with dysplastic urothelium.


Correlation between tumour stage, tumour grade, and immunocompetence in patients with carcinoma of the bladder and prostate. The cellular immunocompetence was examined in 132 patients with transitional cell carcinoma of the bladder and 85 patients with carcinoma of the prostate with reference to the tumour stage and grade. In both groups a significant correlation between tumour stage and grade was found. These two parameters were also correlated with the reduction of immunocompetence. The results are discussed and compared with those in the literature.


Retrospective analysis of 645 simultaneous perineal punch biopsies and transrectal aspiration biopsies for diagnosis of prostatic carcinoma. Transrectal aspiration biopsy of the prostate has been recommended as method of choice for morphological diagnosis of prostatic malignancies. It is less inconvenient to the patient and with hardly any complications. A retrospective analysis of 645 simultaneous perineal punch biopsies and transrectal aspiration biopsies was carried out in order to elicit whether prostatic carcinoma can be detected with transrectal aspiration biopsy as frequently as with perineal punch biopsy, and whether a cytological diagnosis of prostatic cancer is reliable. Carcinoma of the prostate was found in a total of 39.1% either with both techniques or with only one of them. A concordant diagnosis has been obtained in only 25.1%. Prostatic carcinoma was diagnosed more often by perineal punch biopsy  (36.1%) than by transrectal aspiration biopsy (27.7%). However, the diagnosis has been missed by perineal punch biopsy in 2.6% of the cases. Doubtful results and unsatisfactory preparations have been observed more frequently after transrectal aspiration biopsies. Definite 'false-positive' cystological findings have not been detected. Thus, cytological evaluation of aspirated prostatic cells appears reliable. A definite morphological diagnosis can be expected with this technique. Simultaneous application of perineal punch biopsy and transrectal aspiration biopsy can be recommended in order to diagnose prostatic carcinoma with a higher frequency.


Allopurinol in the recurrence prevention of calcium oxalate lithiasis. Since 1973 we have used allopurinol in the prevention and aftercare of recurrent urolithiasis. We give indications for the administration of allopurinol for patients with chronically recurring calcium oxalate lithiasis. Special attention is given to the urinary stone analysis as well as to metabolic disorders as for example hyperuricaemia, hyperuricuria or idiopathic hypercalciuria. In 15 patients with calcium oxalate lithiasis the stone/patient/year ratio could be decreased to 38%. In 19 patients with uric acid/calcium oxalate calculi or alternating stone formations from uric acid and calcium oxalate we succeeded in decreasing this ratio from 1.72 to 0.47 or 27%.


Transrectal ultrasono-tomography.

Transrectal ultrasono-tomography. A relatively simple and painless technique of transrectal ultrasono-tomography is presented. It constitutes a reliable and objective adjuvant to the determination of size and possible infiltration of prostatic and bladder tumours in the human.


Hormone blood levels in patients with prostatic carcinoma and their relation to the type of carcinoma growth differentiation. Human prolactin (HPr), luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, 5 alpha-dihydrotestosterone (DHT), oestrone and oestradiol are measured by radioimmunoassay (RIA) and bilogical prolactin activity additionally by pigeon crop sac assay in plasma of patients with prostatic carcinoma and two control groups. Pigeon crop sac assay is more often positive in cancer patients (50%) than in controls (20%) and HPr distinctly elevated in 20% of the carcinoma patients. Patients are classified according to histological type of tumour: carcinomas with cribriform and/or solid growth show significantly elevated pigeon crop sac activity and higher testosterone, DHT and oestrone levels in plasma than tumours without such growth. Values are discussed with respect to age.


Urological complications in chronic inflammatory diseases of the bowel. In advanced stages, the three most common inflammatory bowel diseases - colonic diverticulitis, Crohn's disease and ulcerative colitis - cause, in some 10% of cases, secondary urological pathology involving either bladder or ureter. Colovesical fistula is found more frequently in diverticulitis and less often in Crohn's disease, which penetrates predominantly from the ileum into the bladder. On the other hand, if uretic stenosis develops it will be caused on the right side by Crohn's disease and on the left by ulcerative colitis. Vesico-intestinal fistulae will close without sequelae after resection of the diseased bowel segment. On the other hand, retroperitoneal ureteric stenosis - despite bowel resection and ureterolysis - will often require nephrectomy if operation is not carried out early enough. Frequent re-checks with infusion excretory urography will help to prevent this. Nephrolithiasis, amyloidosis and contracted bladder are other, though less common, complications of chronic inflammatory diseases of the bowel.


Intracardial blood cyst causing subpulmonary stenosis in congenitally corrected transposition. A patient with congenitally corrected transposition is described. The second sound showed wide fixed splitting of two loud components and hemodynamic and angiocardiographic examinations revealed subpulmonary stenosis. At autopsy a large intracardial blood cyst was found below the pulmonary valves in the outflow region of the morphological left ventricle.


Long-term ECG in ambulatory clinical practice. Analysis and 2-year follow-up of 100 patients studied with a portable ECG tape recorder. A portable tape recorder for long-term ECG monitoring is described. Its light weight (500 g) and small size (138 X 115 X 39 mm) make its usage in routine clinical practice a practical proposition. The most important application has been in the differential diagnosis of Adams--Strokes syndrome. The results from the first 100 patients with a 2-yr follow-up are presented. The importance of GCG recording during  the patients' relevant subjective symptoms is stressed. The mean duration of recording was 2.8 days. In the 28 patients with histories which fitted the symptoms of Adams--Stokes syndrome this diagnosis was confirmed by an arrhythmia recorded simultaneously with the symptoms. In 36 other patients with a similar history the diagnosis was excluded becase of a normal ECG during subjective symptoms. Of the 28 patients with Adams--Stokes sydrome, bradyarrhthmia was the causal factor in 20 patients and these had a pacemaker implanted, whereas the remaining 8 patients had a tachyarrhythmia, which wa treated with antiarrhythmic drugs. The 2-yrs follow-up revealed an improvement and a disappearance of the Adams--Stokes attack in all the patients with an implanted pacemaker. In several of the 36 patients in whom Adams--Stokes syndrome could not be confirmed the syncopal attacks disappeared spontaneously. A large number of arrhythmias, including ventricular and supraventricular tachycardia, 2nd degree AV block and sinus bradycardia were observed during symptom-free intervals in these 36 patients. The introduction of long-term ECG recording routinely in patients with dizziness and syncope of unknown reason has resulted in an increase of the number of patients with a confirmed diagnosis of the Adams--Stokes syndrome, and it has contributed to an increase in the incidence of pacemaker implantation in Malmö from 130 per million inhabitants in 1971 to 220 in 1973, and 1974, respectively.


Wolff-Parkinson-White syndrome type A obscured by left bundle branch block. 2 patients are presented whom the presence of left bundel branch block obscured the signs of the Wolff--Parkison--White syndrome expected with a bypass on the left side of the heart. While it was possible to infer the presence of both disorders from the surface ECGS, electrophysiological studies proved invaluable in defining the precise situation in both cases. When atypical electrocardiographic appearances are observed in the Wolff-Parkinson-White syndrome, careful investigation may reveal the cause to be bundle branch block on the same side of the hear as the anomalous pathway. In both patients there was in addition evidence of sinoatrial disease, which was associated with the occurrence of arrythmias, and this was also confirmed with intracardiac studies.


Auto-interference by a demand pacemaker. Immediately after implantation of a unipolar demand pacemaker, inappropriate inhibition accompanied by instability of the ECG baseline became apparent. Later, higher frequency artifact appeared and the pacemaker fired intermittently in its fixed rate mode, indicating the sensing of severe interference. These phenomena were subsequently found to have resulted form ecposure on the pacemaker surface of a connection between the negative side of the battery and the electronic components.


Paradoxical potassium excretion in response to aldosterone antagonists. The activity of two aldosterone antagonists in reversing the urinary electrolyte changes induced by the mineralocorticoid fludrocortisone was examined in healthy subjects. Between 2-10 h after treatment there were dose-related increases in sodium excretion and the urine log 10 Na/K ratio, but no significant changes in urine volume, potassium concentration, or potassium excretion. Between 12-16 h after treatment there were dose-related increases in urine sodium excretion and the log 10 Na/K ratio. Unexpectedly, there were significant dose-related increases in potassium excretion despite significant dose-related reductions in urinary potassium concentration. The paradoxical increases in potassium excretion were attributed to dose-related increases in urine volume in the same period. These observations may explain the increased potassium excretion occasionally observed during clinical use of aldosterone antagonists and suggest that potassium excretion in man is influenced by the urine flow rate.


Exercise-induced asthma and doxantrazole. In a double-blind cross-over trial in 13 patients with exercise-induced asthma, doxantrazole 200 mg taken orally 1 hour before a standardized exercise test involving stair climbing failed to block post-exercise bronchoconstriction. In an open assessment study with the same patients, increasing the doxantrazole dose to 400 mg did not affect post-exercise bronchoconstriction.


Acute effects of acetylsalicylic acid in patients with chronic renal insufficiency. The effect of acetylsalicylic acid (ASA) in patients with renal insufficiency has been examined. In one investigation (A), in patients with a mean GFR of 23.0 ml/min the acute effects of ASA 750 mg i.v. (lysine-ASA 7.5 ml) and 0.9% NaCl 7.5 ml on renal water and solute output and on the clearance of inulin, creatinine and PAH were compared. In another (B) the effects of simultaneous administration of ASA 750 mg or 0.9% NaCl 7.5 ml i.v. with an infusion of furosemide 250 mg were investigated in six patients (mean GFR 12.9 ml/min) in a cross-over study. In study A there was a significant fall in urinary sodium excretion within the first 15 min after ASA administration, with a maximal decrease to 21% of the control period. Urine flow fell to 35%, osmolal clearance to 41%, inulin clearance to 54% and PAH clearance reabsorption of sodium increased. The effect of ASA lasted for 2-6 h. The mean salicylic acid concentration during the first two hours after ASA administration was 60.0 mug/ml, and the mean protein bound salicylic acid (SA) was 70.4%. There was no effect of placebo (0.9% NaCl 7.5 ml) on renal function. Pretreatment with ALA 750 mg i.v. attenuated the diuretic effect of furosemide 250 mg, and reduced creatinine clearance significantly within 0-2 h after drug administration.


Acute effects of acetylsalicylic acid on renal function in normal man. The acute effects of therapeutic doses of acetylsalicylic acid (ASA) on renal water and solute output, and the possible interaction of ASA with the diuretic effects of furosemide, have been studied in a double blind double cross over study in healthy human subjects. There was a significant decrease in 24 h sodium excretion and Na/K ratio in urine in the ASA-treated subjects. The effect of ASA on urinary sodium excretion was most prominent during day time (8 a.m.-10 p.m.) and on days with low sodium intake, as confirmed by control sodium excretion and plasma renin activity. A decrease in urine volume and an increase in tubular reabsorption of free water were caused by ASA, the antidiuretic effect being most marked at night (10 p.m.-8 a.m.). No action of ASA on the effect of furosemide on urinary sodium excretion was found. Creatinine clearance remained unaltered by ASA treatment, and ASA did not interfere with the increase in urinary creatinine excretion after furosemide treatment.


Monoamine metabolites in cerebrospinal fluid during treatment with clonidine or alprenolol. Lumbar cerebrospinal fluid (CSF) concentrations of the major metabolites of noradrenaline (4-hydroxy-3-methoxyphenyl glycol, HMPG), serotonin (5-hydroxyindoleacetic acid) and dopamine (homovanillic acid) were measured before and during the administration of clonidine or alprenolol to hypertensive patients. The noradrenaline receptor stimulant clonidine significantly decreased the CSF level of HMPG, but there was no consistent change in the concentration of serotonin or dopamine metabolites. Patients on alprenolol showed no change in the levels of these metabolites in CSF.


Carbamazepine and its 10,11-epoxide in children and adults with epilepsy. The plasma concentrations of carbamazepine and its 10,11-epoxide were measured in 37 children and 13 adults with epilepsy during maintenance therapy. The children formed relatively more of the epoxy metabolite than adults. The daily dose, expressed as mg/kg or mg/m2, showed a statistically significant correlation with blood level in children, but not in adults. The cerebrospinal fluid concentrations of carbamazepine and carbamazepine-10,11-epoxide in nine children were 33 and 41 per cent, respectively, of the concomitant plasma level.


Plasma levels and half lives of thioridazine and some of its metabolites. II. Low doses in older psychiatric patients. Plasma-levels of thioridazine, mesoridazine, sulphoridazine and two other metabolites were determined in ten older chronic psychotic patients on thioridazine therapy. The plasma-level before the morning dose of thioridazine was the most reliable parameter for clinical studies. An intra-individual relationship between lower doses of thioridazine and plasma-levels was found. The percentage contribution of psychoactive compounds to the total sum of "thioridazine plus metabolites" ranged from 43-74%. The mean "early disappearance half-life" of thioridazine was 5 hours, and its mean "late disappearance half-life" was 26 hours.


Fluorometric determination of hydroflumethiazide in human plasma and urine after its oral administration. A spectrofluorometric method for determination of hydroflumethiazide in human plasma and urine has been developed. The limit of detection was 10 ng/ml plasma and 100 ng/ml urine. The plasma concentration of hydroflumethiazide was determined for 9-11 hours and excretion in urine for 24-37 hrs after oral administration of about 1 mg/kg body weight to 7 subjects. Plasma half life in healthy subjects was 1.9-2.1 h, and 2.7-8.6 h in patients during the period 4-9 hrs after dosing. Cumulative excretion in urine was 67-79% of the dose during 31-37 hrs in 6 subjects; one patient with renal disease was found to excrete only 25.8% of dose during 24 hours. Renal clearance of hydroflumethiazide was higher in the healthy subjects (0.29-0.44 1 h-1 kg-1) than in the patients (0.040-0.15 1 h-1 kg-1). Plasma half life of hydroflumethiazide was not closely correlated with renal clearance of the drug, which suggests that other factors may play a role in determining plasma half life.


The ambiguity of adverse drug reactions. Three clinical pharmacologists independently evaluated 500 untoward clinical events reported by physicians as adverse drug reactions (ADRs). They often disagreed with the reporting physicians and with each other. They judged 14.4-28.2% of the events to be definite, 26.4-38.0% probable, 21.4-31.0% possible and 15.8-28.2% of the events to be definite, 26.4-38.0% probable, 21.4-31.0% possible and 15.8-28.2% unlikely ADRs. In their opinion 19.1-32.4% of the drugs blamed were definitely, 29.8-34.4% probably, 24.9-36.7% possibly and 8.4-14.5% not responsible for the adverse reactions. Evaluators disagreed among themselves about the drug most likely to have been responsible in 36.4% of the events, about ADRs causing hospital admission in 56.8%, about severe ADR morbidity in 55.8%, about ADR prolongation of hospitalization in 67.3% and ADR prolongation of hospitalization in 67.3% and about ADR contribution to death in 71.0%. The divergence of judgements suggests that suspected ADRs are usually ambiguous clinical events, and that incidence, severity, medical consequences and cost of ADRs can only be estimated.


Reserpine and breast cancer in women in germany. Exposure to reserpine was compared in 181 women interviewed prior to biopsy and found to have breast cancer and 307 women found to have a benign disorder of the breast. The age-adjusted relative risk of breast cancer in those who had taken reserpine was 0.6 (95% confidence limits: 0.4 and 1.1). When the 181 breast cancer patients were compared with a second control group of 101 women with a benign condition requiring surgery, the relative risk was 0.9 (95% confidence limits: 0.4 and 1.7). Neither long-term exposure nor its timing, gave any evidence of an association with breast cancer. The findings in this study do not support the hypothesis that rauwolfia derivatives initiate or promote breast cancer.


Effect of lisuride and other ergot derivatives on monoaminergic mechanisms in rat brain. The effects of the ergot derivative lisuride hydrogen maleate on synthesis, turnover and receptor activity of monoamines were compared with those of LSD, methysergide, yohimbine and methiothepin. In the dopamine (DA) rich areas, c. striatum and mesolimbic forebrain, lisuride, (30-100 mug/kg) decreased the rate of dopa formation after inhibition of the aromatic amino acid decarboxylase with NSD 1015, 100 mg/kp i.p. After axotomy of the ascending monoaminergic fibers, lisuride (0.5 mg/kg) antagonized the accumulation of dopa in c. striatum and mesolimbic forebrain even on the lesioned side; haloperidol effectively counteracted this lisuride-induced decrease in dopa formation. In the predominantly noradrenaline (NA)-innervated neocortex, lisuride in doses of 0.3 and 1.0 mg/kg increased dopa accumulation. Methysergide, yohimbine and methiothepin also stimulated tyrosine hydroxylation. After inhibition of catecholamine synthesis with alpha-methyl-p-tyrosine methylester HCl, lisuride decelerated DA disappearance and accelerated NA disappearance. 3-Methoxytryramine (3-MY) accumulating after inhibition of monoamine axidase with pargyline HCl (100 mg/kg) was used as an indicator of DA release. Lisuride and LSD, 50 mug/kg administered twice, reduced 3-MT formation while methysergide, 50 mg/kp i.p. had no effect on 3-MT accumulation. All compounds except methiothepin reduced 5-hydroxytryptophan (5-HTP) accumulation in whole brain after NSD 1015. In addition, lisuride caused an increase in 5-hydroxytryptamine (5-HT) and a decrease in 5-hydroxyindole acetic acid concentration. In spinal reserpinized rats, lisuride was indistinguishable from LSD in inducing extension and athetoid movements of the hind legs. The data support the view that lisuride stimulates pre- and postsynaptic DA and 5-HT receptors and suggest that lisuride blocks NA receptors in the central nervous system.


Possible storage of (+)-amphetamine in catecholaminergic terminals of the striatum and brainstem. 6-Hydroxydopamine, given intraventricularly, did not affect the high concentrations of (+)-amphetamine present in the rat striatum and brainstem 1 h after its administration but considerably reduced the small amounts of (+)-amphetamine remaining after 5 h. In contrast, 5,6-dihydroxytryptamine did not modify the (+)-amphetamine concentrations at the times tested. These findings suggest that (+)-amphetamine might be stored in the catecholaminergic but not in the serotonergic central terminals.


Effect of dexamethasone on monoamine oxidase inhibiton by iproniazid in rat brain. Chronic (6 days) dexamethasone administration caused a slight decrease of rat brain MAO enzyme activity which was reflected by lower levels of 14C-homovanillic acid (HVA) and increased levels of 14C-3-methoxytyramine (3MT) following intracisternal injections of 14C-dopamine (DA). Opposite results with dexamethasone were obtained in iproniazid (MAO-inhibited)-treated rats. In these animals, brain MAO enzyme activity was significantly increased by dexamethasone. This effect increased with the duration of dexamethasone treatment and appeared to be dose dependent. In the brain areas tested (hypothalamus, midbrain, cerebellum, pons and medulla, olfactory, rest of brain) increases of MAO enzyme activity were also indicated by lower levels of 14C-3MT and increased levels of 14C-HVA formed from intracisternally injected radiolabeled DA. Treatment with other glucocorticoids (16alpha-methyldichlorisone, 16beta-methylprednisone and prednisolone) had a similar effect on 14C-DA metabolism. On the other hand, desoxycorticosterone, progestone, estradiol and testosterone, did not exhibit this property. The data indicate that chronic glucocorticoid treatment may have a slight inhibitory effect on brain MAO and also has the ability to partially reverse or antagonize the inhibition of MAO caused by iproniazid.


Increase of free arachidonic acid by furosemide in man as the cause of prostaglandin and renin release. Arachidonic acid (C20 :4) plasma renin activity (PRA), PGF2alpha, and sodium excretion were determined before and after furosemide in men. C20 : 4 and PRA increase (p less than 0.005) within 10 min after furosemide, PRA then decreased again whereas C20 : 4 levels remained elevated. Maximal exretion of PGF2alpha and sodium occurred 30-60 min after furosemide. Indomethacin prevented the rise of C20 : 4, PRA and PGF2alpha after furosemide, leaving sodium excretion unaltered. The release of C20 : 4 is assumed to be the primary mechanism of furosemide to increase PG biosynthesis and renin release.


Decreased anticonvulsant activity of carbamazepine in 6-hydroxydopamine-treated rats. An intraventricular injection of 6-hydroxydopamine, which produced a marked decrease of catecholamines in the forebrain, significantly lowered the electroconvulsive threshold in rats. The anticonvulsant effect of carbamazepine was also significantly reduced in the animals treated with 6-hydroxydopamine. The results are consistent with the hypothesis that brain catecholamines may play an important role in seizure susceptibility as well as in the anticonvulsant activity of carbamazepine in rats.


Effect of ouabain on reinnervating mammalian skeletal muscle. The resting membrane potential (RMP) of denervated mammalian muscle fibers increases when reinnervation occurs. This recovery of RMP is temporally associated with the return of a ouabain-sensitive fraction of the RMP. The data suggest that the activity of an electrogenic pump within the sarcolemma is, either directly or indirectly, under neurotrophic regulation.


Somatosensory function and cortical unit activity in cats with only dorsal column fibers. Cats received lesions that transected the entire thoracic cord except for partial sparing of the dorsal columns. The cats were required to discriminate the side on which they were touched, the size of simultaneously presented discs, or the direction their fur was stroked to obtain food reward. All cats found by anatomical and/or electrophysiological criteria to have any functional continuity in the dorsal columns were able to master the first of these tasks; some responded above chance on the second. Performance was at chance on blank trials, and cats with complete cord transection failed to discriminate. Lesioned cats did not orient or otherwise react to any nonrewarded stimulus below the level of the lesion. A total of 532 units were recorded under light barbiturate anesthesia from the hind paw projection near the tip of the ansate sulcus in these and other similarly prepared cats. Three-fourths of the units found before and acutely after the cord lesions were made were driven by hind limb stimulation. Only 27% of the units recorded 10 or more days afterwards could be driven. Of these driven units, 15 (38%) responded to foreleg stimulation, 13 exclusively so. No such units were found in intact or acutely lesioned cats.


Discharges of superior colliculus neurons during head and eye movements of the alert cat. 452 single neurons from the superior colliculus were recorded in awake and non-paralysed cats. 75 neurons were obtained from cats with unrestrained horizontal head movements. 228 neurons remained unaffected by saccadic eye movements. Eye movement related discharge followed the onset of saccades in 156 neurons either only in the presence of a visual pattern (92 neurons) or in darkness, too (64 neurons). The latter reaction type probably depends on eye muscle afferents. In 48 neurons eye movements related activity preceded the onset of eye movements. 12 neurons eye movement related activity preceded the onset of eye movements. 12 neurons fired in synchrony with eye movements of any direction (type I). 30 neurons were excited during contralaterally directed eye versions within or into the contralateral head related hemifield. They were inhibited when the eyes moved within or into the ipsilateral head related hemifield (type II). 6 neurons with constant maintained activity during fixation were inhibited by ipsilaterally directed saccades, but remained unaffected by contralateral eye movements. Head movement related discharge followed the onset of head movements in 20 neurons only in presence of a visual pattern and also in darkness in 6 neurons. Ipsilateral head movements or postures strongly suppressed maintained activity and visual responsiveness of some neurons. 15 neurons discharged in synchrony with and prior to contralateral head movements. Ipsilateral head movements inhibited these neurons. Activation or inhibition were usually related to movement and to posture, exceptionally to movement or to posture. Electrical stimulation of recording sites of these neurons through the recording microelectrode elicits contralateral head movements.


The rubro-bulbospinal path. A descending system known to influence dynamic fusimotor neurones and its interaction with distal cutaneous afferents in the control of flexor reflex afferent pathways. The inhibitory effect of electrical stimulation in the near-rubral region on polysynaptic segmental as well as ascending pathways activated by the flexor reflex afferents (FRA) in hind limb nerves was studied in chloralose anaesthetized cats. The effective stimulating region totally coincided with the one from which a D zone climbing fibre response may be elicited in the contralateral cerebellar cortex. The descending path was dependent upon an intact dorsolateral spinal funiculus, where also a characteristic volley could be recorded with a surface electrode on short train central stimulation. The suppressive action on the transmission through the FRA pathways was evoked in the absence of a lower lumbar dorsal root potential, and it was concluded that the effect was exerted by postsynaptic inhibition. It was suggested that this descending path, the effects of which resemble those elicited from the dorsal reticulospinal system, is identical to the rubro-bulbospinal path, previously known to influence dynamic fusimotor neurones. The transmission through the FRA pathways was also suppressed by conditioning stimulation of ipsilateral, low threshold distal cutaneous afferents. The time course of this effect was the same as that with central conditioning stimulation. Facilitatory interaction was revealed with double conditioning and it was suggested that the descending path and the distal cutaneous afferents converge upon a common group of interneurones, which postsynaptically inhibit an early (possibly the first one) interneurone in the FRA pathways. As low threshold distal cutaneous afferents supply the primary peripheral input via climbing fibres to the cerebello-cortical D zone, it was concluded that the different stimuli (central or peripheral) which activate a common group of inferior olivary neurones destined for the D zone also activate a common group of segmental inhibitory interneurones. The results are discussed in relation to current concepts of segmental motor control, and it is suggested that the mechanisms studied could be involved in the regulation of stepping.


Movement adaptation in the visual evoked response. Visual evoked responses to sinusoidal movement before and after adaptation to linear movement were compared. Fourier analysis indicated that the responses evoked by sinusoidal movement were insensitive to direction of movement. Following adaptation, the harmonics describing the waveforms changed in magnitude and phase, and showed an asymmetry of waveshape not evident before adaptation. The data are consistent with the existence of movement-sensitive cells within the intact visual system which change in response characteristics as a result of adaptation to linear movement.


Spatial properties of binocular neurones in the human visual system. The spatial properties of human binocular mechanisms were investigated using the technique of subthreshold summation. Isolation of binocular mechanisms was achieved by means of interocular stimulus presentation. The contrast detection threshold for a sinusoidal test grating viewed by one eye was found to be reduced by a subthreshold grating of the same spatial frequency and orientation seen by the other eye. The interaction between the gratings was approximately linear. Threshold increased as the spatial frequencies or orientations of test and subthreshold gratings were made increasingly different. Spatial stimulus specificities measured in this way were as great for interocular presentation as for simultaneous monocular presentation. The results suggest that human contrast sensitivity for gratings may depend upon binocularly-activated neurones similar to those found in cat and monkey visual cortex.


Dietary-induced overeating in experimental animals. Modifications in dietary conditions can result in small, but consistent, increases in caloric intake which over time accumulate to substantial increases in body weight. Allowing rats access to either high fat diets or a variety of highly palatable foods can lead to obesity. Recent experiments also have shown that providing access to sweet carbohydrate solutions in addition to the complete diet can cause weight gains in normal neurologically intact adult rats. Although animals increase consumption of carbohydrate on this dietary regime, they continue to maintain minimum requirements for other dietary nutrients. It appears that increasing the palatability of the diet can lead to overeating, but cannot induce the rat to select a diet that is deficient in protein, fat, vitamins, or minerals. Further exploration of the conditions associated with dietary-induced overeating in animals may provide insights into conditions related to obesity in man.


Isolated perfused lung--substrate utilization. Lung metabolism has been extremely difficult to determine in vivo primarily because the lung is overwhelmed by a great blood flow that generally makes the Fick principle inadequate. Largely for reasons such as this, investigators have had to rely on in vitro preparations. The isolated perfused lung has the apparent advantage of being similar to the lung in vivo when compared with other preparations. For instance, there is evidence that the capillary bed of the lung may alter substrates and influence their subsequent metabolism. Substrates have contact with the capillary endothelium in isolated perfused lungs but not to tissue slices, homogenates, or isolated cells. Our studies indicate that precursors of saturated phosphatidylcholine may include lipids, which are hydrolyzed in the capillary of the isolated perfused lung and thus become substrates such as free fatty acids, etc. However, tissue slices do not use the esterified lipids to the same extent, presumably because in this preparation the enzymes in the capillary endothelium do not have contact with the esterified lipids. Substrate utilization of the isolated perfused lung may be considerably altered by inflation of the lung or by pulmonary edema. Although glucose utilization and palmitate oxidation by the isolated perfused lung and by tissue slices of the rat lung are very similar, if the isolated perfused lung develops pulmonary edema, glucose utilization increases by nearly 100%. This phenomenon is apparently not due solely to fluid in the airspaces because in control studies with fluid added into the airways the glucose utilization did not increase to the degree observed with edematous lungs. Lung distention is associated with increased glucose consumption but marked distention is also associated with pulmonary edema. The effect of lung distension may be a direct effect or it may be secondary to the pulmonary edema.


Nutrient transport systems in dog brain. The cerebral circulation of the dog was isolated and an extracorporeal system was used to perfuse the brain with diluted blood. Transport of blood-borne substances was investigated by measurement of arteriovenous (a-v) differences (net transport) and by the indicator dilution method (unidirectional influx). The net movement of amino acids between blood and brain was qualitatively and quantitatively different when measured by whole blood a-v differences than when measured by plasma a-v differences. This result suggests that erythrocytes are involved in the movement of amino acids between the blood and brain, a finding similar to that observed in other vascular beds (Elwyn et al. Am. J. Physiol. 222: 13338 1972). The transport of 13 different blood-borne substances was investigated at normal and elevated solute concentrations using the indicator dilution method. Saturable processes were observed for the unidirectional transport of tyrosine, tryptophan, L-dopa, free palmitic acid, adenine, thymine, adenosine, and guanosine. However, the blood-brain interface did not appear to contain a carrier for dopamine, folic acid, cyanocobalamin, guanine, hypoxanthine, and free oleic acid.


Effect of starvation on muscle glucose metabolism: studies with the isolated perfused rat hindquarter. Studies in man and experimental animals suggest that the metabolism of glucose by skeletal muscle is depressed during starvation. To investigate the basis for this, the effect of starvation on the uptake and disposition of glucose in skeletal muscle was studied in the isolated perfused rat hindquarter. In contrast to earlier work carried out in heart, neither glucose uptake, whether stimulated by insulin or exercise, nor glycolysis were depressed by 48 hr of starvation or by perfusion of the hindquarter with acetoacetate, palmitate, or octanoate. Glucose oxidation, assessed from the oxidation of 1-[14C]lactate, was depressed by approximately 75% in starved rats and by 30% in fed rats perfused with acetoacetate. Exercise increased lactate oxidation 10-fold in both fed and starved rats; however, the relative difference between the groups persisted. In general, changes in lactate oxidation were paralleled by changes in the activity of pyruvate dehycrogenase (active form). The data suggest that glucose metabolism in skeletal muscle is inhibited during starvation at the step of pyruvate oxidation and that this inhibition persists during exercise. The also suggest that the diminution of glucose uptake that occurs in skeletal muscle of intact organisms during starvation may not be related to the presence of high concentrations of free fatty acids and ketone bodies.


Vascular perfusion of rat small intestine: metabolic studies with isolated and in situ preparations. Two different preparations of vascularly-perfused rat small intestine are described and their application to metabolic studies in this tissue illustrated. The first, a totally isolated, blood-perfused intestinal preparation, has an apparent requirement for glucocorticoid and norepinephrine. Lymph is produced and collected, permitting studies of fat transport and lipoprotein biosynthesis. Results indicate that intestine synthesizes some, but not all, of the proteins associated with chylomicrons and other lipoproteins of intestinal lymph. The isolated intestine from 260-g rats metabolizes 75 mumoles of circulating glutamine per hour, mostly to CO2, lactate, citrulline, proline, alanine, and ammonia. In a second preparation, all venous blood is collected from a short, isolated intestinal segment in situ with intact arterial blood supply. Adaptations allow the quantitative determination of rates of uptake and metabolism of substances from both blood and lumen. Results indicate that over 36% of CO2 produced by intestine of fasted rats is derived from plasma glutamine and only 8% from glucose. Virtually all glutamate and aspartate and significant amounts of glutamine and arginine absorbed from the lumen are also metabolized. The resulting metabolic products were quantitated and include CO2, lactate, proline, citrulline, alanine, and glucose.


Metabolism of cyhlomicrons in perfused adipose and mammary tissue of the rat. Perfused adipose and mammary tissues of the rat were used to study the role of capillary endothelium and lipoprotein lipase in the metabolism of chylomicrons. Triacylglycerol of chylomicrons was hydrolyzed stepwise during uptake by both tissues, first to di- or monoacylglycerol and fatty acid, and then to glycerol and fatty acid. Cytochemical studies in perfused adipose tissue showed that lipolysis occurred at the luminal surface of capillaries, within vacuoles and microvesicles in the endothelium, and between the endothelium and pericytes. Radioautography of perfused lactating mammary tissue showed that acyl and glyceryl moieties derived from chylomicron triacylglycerol were both incorporated into milk lipid droplets. Perfused lactating mammary tissue also took up cholesterol from chylomicrons. There was marked reduction of lipoprotein lipase activity and uptake of both triacylglycerol and cholesterol in perfused mammary tissue of hypophysectomized lactating rats. It is proposed that uptake of triacylglycerol by extrahepatic tissues requires the direct action of lipoprotein lipase, while uptake of cholesterol is dependent on reduction of the triacylglycerol core volume, resulting from action of the enzyme on the core and uptake of lipolytic products by the tissue.


Rumen bacteria: interaction with particulate dietary components and response to dietary variation. The bovine rumen resembles many other ecosystems in that its component bacterial cells are universally surrounded and protected by extracellular structures. The most common form of these structures is a fibrous carbohydrate slime that extends away from the cell and may mediate the attachment of the bacterium to a surface. This attachment is relatively specific and it may occur at the surface of the rumen epithelium or on the cell walls of a specific tissue within the plant-derived food of the animal. The production of the extracellular slime is under nutritional control and slime may be overproduced when soluble carbohydrates are available in high concentration. This overproduction results in cell-cell adhesion among the rumen bacteria with the eventual formation of slime-enclosed microcolonies and, in extreme cases, the generation of sufficient viscosity to cause feedlot bloat.


Ruminal microbial yields: factors influencing synthesis and bypass. Bacterial growth (protein production) in the rumen is typically limited by anaerobic energy supply. But the mass of bacteria produced per mole of ATP (YATP) varies markedly with turnover or growth rate of bacteria, availability of cell components, accumulation of ash or starch, and intraspecies transfer of reducing equivalents. Increased turnover rate of rumen contents appears to enhance bacterial protein production, increase ruminal acetate and methane production and increase bypass of fiber and concentrate components of the ration.


Estimating rapid changes in the rates of glucose production from glycogenolysis and recycling through lactate. In order to obtain independent estimates of changes in the rate of glucose production from glycogenolysis from the glucose carbon recycled throught lactate, previously validated nonsteady tracer methods are used. Changes in glycogenolysis are estimated by calculating the rate of appearance of a label ([6-3H]glucose) uniformly distributed through glycogen and simultaneously the rate of recycling of 14C is measured under nonsteady-state conditions. It is indicated that these results can be extended to the determination of absolute rates of glucose production from glycogen breakdown and gluconeogenetic precursors.


The turnover and conversion to glucose of alanine in newborn and grown dogs. The extent of transfer of carbon atoms from alanine to glucose was examined in pups and grown dogs. [U-14C]Alanine and [2-3H]glucose were injected intravenously and by using the SAAM-26 program a compartmental model was formulated from the tracer data to quantify the kinetics of the alanine- and glucose-carbon system. A 3-compartment model was necessary to describe the alanine-carbon kinetics. Strict identification of these compartments with physiological counterparts was not possible. The overall transport (turnover) rate of alanine-C was found not to change significantly with age when calculated on the basis of body weight. Carbon atoms from alanine reach glucose by three identifiable pathways of different speed. The kinetics suggest that the different pathways are at least in part due to differences in the metabolism of the three carbon atoms of alanine. In pups less than 11 days of age 45-49% of the carbon atoms leaving the alanine subsystem enter glucose; in adults it was 70%. In young pups 18-19% of glucose-C utilized was derived from alanine-C, whereas in adults, 62%. Accordingly a smaller percentage of carbon atoms utilized as alanine appears in glucose and a smaller percentage of glucose-C is derived from alanine in pups than in grown dogs.


Measurement of glucose recycling and liver glycogen synthesis in mice using doubly labeled substrates. Tracer experiments have been carried out using gorging and nibbling mice to study several related aspects of carbohydrate metabolism: 1) inhibition of gluconeogenesis shortly after animals ingest a glucose-rich meal; 2) the extent to which dietary glucose carbon is recycled by way of 3C compounds after dietary glucose is absorbed; and 3) recycling of glucose by exchange between free and a hypothetical, "bound" glucose pool. Fasted, gorging mice were allowed to eat 120 mg [U-14C,6-T]glucose (58% glucose diet) in 4 min. Plasma glucose-C specific activity rapidly reached that of the dietary glucose-C. Superficially, this suggested nearly complete inhibition of hepatic gluconeogenesis. However, plasma [6-T]glucose, glycogen-[14C, 3H]glucose analyses, and [14C]-glycerol conversion to glucose showed that hepatic gluconeogenesis continued during alimentary hyperglycemia. Half of liver glycogen seemed to be formed from a hepatic G-6-P pool that was never labeled. Indirect kinetic evidence of a large, bound exchangeable glucose pool was presented. Since no direct evidence of such a pool has been obtained, the possibility is raised that a serious artifact of the tracer technique exists or else some unconventional model of carbohydrate metabolism is required to explain our data.


Management of hyperstimulation syndrome. A case of hyperstimulation syndrome secondary to Pergonal therapy is presented. Successful management was based principally on severe sodium and fluid restriction without the use of volume expanders. The rationale for this therapeutic approach is presented and discussed. Although this iatrogenic disease should be virtually eliminated with the monitoring of daily urinary estrogens, severe hyperstimulation may still occur as a result of laboratory error.


Meiotic figures in the testicular biopsy of subfertile males. The phases and sequence of meiotic division are herein reviewed and illustrated as they occur in the human testis. Normally all phases of meiosis can be identified, and arrest at a phase is indicative of faulty spermatogenesis. The correlation of these arrest patterns with the clinical course and response to clinical therapy is now in progress. These meiotic studies can be undertaken in any laboratory with cytogenetic capabilities, and this study is intended to serve as a base line illustration of the normal.


Effect of intrauterine estriol on reproductive function in the rabbit. The effect of intrauterine estriol on spontaneous ovulation, ovum fertilization, blastocyst development, and blastocyst implantation in rabbits has been investigated. Estriolreleasing intrauterine capsules constructed of biocompatible polymer were implanted in one uterine horn of adult New Zealand White female rabbits, while placebo-containing capsules were implanted into the contralateral horn. The animals were artificially inseminated and ovulated or mated to fertile bucks. The does were killed and their reproductive tracts were examined 54 hours, 6 days, and 10 days after ovulation-inducing injection or after coitus. The results indicate that intrauterine estriol released at a steady state rate of 1.25 mug/day effectively inhibits blastocyst development and implantation. This contraceptive effect was clearly local, since implantations in the contralateral,placebo-bearing horn were not inhibited. Furthermore, the same dose of estriol, when released systemically from a subcutaneously placed capsule, had no effect on implantation. Intrauterine estriol appeared also to have no effect on spontaneous ovulation or ovum fertilization.


Haemodynamic effects of dobutamine in patients with coronary artery disease. The haemodynamic effects of the infusion of dobutamine in dosages of 2-5, 5-0 and 10-0 mug/kg/min were studied in twelve patients with coronary artery disease. At the lowest dose, infusion of dobutamine caused a significant increase in cardiac output; greater increases occurred with doses of 5-0 and 10-0 mug/kg/min. The means by which the myocardium produced the increase in cardiac output differed according to the dose of the drug; 2-5 and 5-0 mug/kg/min produced an increase in stroke volume without any significant variation in the heart rate. At the highest dose rate, an increase in heart rate made a distinct contribution to the overall increase in output. The ventricular dp/dt max. increased at all three dosages. Blood pressure, pulmonary arterial and capillary pressure, and left ventricular and diastolic pressure did not show any significant change. The separation of the inotropic and the chronotropic effects of dobutamine according to the infused dose appears to have extremely interesting clinical implications.


Maprotiline (Ludiomil) in depression: a report of a monitored release study of 10,000 patients in general practice. In a three-week study of 10,000 general practice patients with depressive illness, a single dose of maprotiline 75 mg at night was effective therapy in three-quarters of the patients who completed the study. At this dose the drug was well tolerated, and troublesome side-effects presented in only a small percentage of patients. Drowsiness was the most commonly reported side-effect, and the main reason for withdrawal of treatment. The drug was acceptable to most physicians, and in this study was used in a wide range of patients and over a broad spectrum of depressive illness.


The mechanism of action of a normophasic oral contraceptive 'Fysioquens'. Six women with normal menstrual cycles were treated over six cycles with Fysioquens, a normophasic contraceptive consisting of 7 tablets of ethinyloestradiol (EE) 0-050 mg and 15 tablets containing 1-0 mg lynestrenol + 0-050 mg EE. In the 1st, 3rd and 6th treatment cycles and in the cycles before and after treatment, hormonal assays and other tests were carried out to determine whether ovulation had occurred. No treatment cycles showed any signs of ovulation, but it did occur in the pre-and post-treatment cycles. Blood pressures and liver function tests remained normal, but two of the six volunteers showed a slight increase in body-weight.


An assessment of the bacteriological activity of Locabiotal (Fusafungine) --a non-systemic antibiotic. Various tests were performed to establish the bacteriological potency of Locabiotal, an aerosol for local application to the nose and throat containing fusafungine. Using impregnated filter paper on a plate of solid sensitivity media considerable inhibition of the growth of common pathogens was shown. Plates of nasal swabs from adults and buccal swabs from children showed similar inhibition. Further tests after the application of the spray to the nose and throat indicated that inhibition lasts for at least eight hours.


A general practice assessment of Locabiotal in the treatment of upper respiratory tract infections. An open clinical trial of Locabiotal, in which the antibiotic, fusafungine, is presented in a micronized aerosol was performed in general practice. One hundred and ten patients suffering from upper respiratory infections without associated hyperpyrexia or toxic symptoms were treated for 7 days. There was a marked improvement in the symptoms rated during the first 48 hours. After the full course of treatment 37 patients regarded its effect as excellent and 48 as good.


Comparative effect of food on absorption of ampicillin and pivampicillin. The effect of food on the absorption of simultaneously ingested ampicillin or pivampicillin was compared in a crossover study in eight healthy volunteers. The absorption of both ampicillin and pivampicillin was delayed by simultaneous food intake as judged by serum concentration and urinary excretion of ampicillin. The total absorption of ampicillin, but not that of pivampicillin was decreased by simultaneous food intake as indicated by the are under the serum ampicillin concentration-time curves and by 24 hours urinary excretion of ampicillin. The excretion of ampicillin into urine was about 30% of the dose when ampicillin was ingested with water into an empty stomach and about 20% when ingested with food. The respective excretion of ampicillin following the ingestion of pivampicillin was about 60% of the dose taken either with or without food.


Purification of oestradiol receptor from human uterus by affinity chromotrgraphy. The oestrogen receptor from human myometrium has been extensively purified by affinity chromatography and isoelectric focusing. The latter step is necessary to remove contaminating sex-steroid-binding globulin. The unpurified 3.7-S cytoplasmic receptor has a molecular weight of 41,000, a Stokes radius of 27.0 A and a frictional ratio (f/f0) of 1.19; the KD (4 degrees C) for [3H]oestradiol-17 beta was 1.03 X 10(-10) M. After purification, the molecular weight was 30,000, the Stokes radius 23.6 A, frictional ratio 1.15 and isoelectric point 6.15.


The presence of an estradiol binding component in cytosol from immature rat epididymides. Cytosol prepared from epididymides of sexually immature (21-23-day-old) rats contains a macromolecular binding component for estradiol-17 beta. This binding moiety sediments as an 8-S species on 5-20% sucrose gradients containing 0.01 M KCl. Under conditions of high ionic strength (0.4 M KCl) the 8-S peak of estradiol binding is shifted to the 4-S region, suggesting dissociation of receptor aggregates. Time-course studies indicated that binding equilibrium was essentially achieved after 2 hours incubation at 0 degrees C. Although unlabeled estrone and estriol are capable of inhibiting [3H]estradiol binding to epididymal cytosol, they are less effective than unlabeled estradiol. Unlabeled 5 alpha-dihydrotestosterone (5 alpha-DHT) at a 100-fold molar excess did not cause a statistically significant inhibition of [3H]estradiol binding. Unlabeled estrogens, but not unlabeled 5 alpha-DHT or cortisol (at the concentrations used), were capable of displacint [3H]estradiol from its binding sites. The dissociation of [3H]estradiol from the binding component is very slow, with half-time of dissociation being greater than 16 hours. The epididymal estrogen binder is saturable at low concentrations of ligand. The dissociation constant was of the order of 10(-11)M and the concentration of binding sites was approximately 10(-14) mol/mg protein. This estrogen binder has the characteristics which are usually attributed to steroid receptors and is clearly different from the testicular androgen-binding protein and the epididymal androgen receptor.


Suppression of 20 alpha-hydroxysteroid dehydrogenase activity in cultured rat luteal cells by prolactin. 20alpha-Hydroxysteroid dehydrogenase (20 alpha-SDH) activity increases in the cycling corpus luteum of the rat, beginning at 14.00 h on the day of diestrus, but remains low in corpora lutea of pregnancy throughout the first 19 days of gestation. When cells derived from 7-day-old corpora lutea of pregnant rats were cultured for 7 or 12 days, there was a spontaneous rise in 20 alpha-SDH activity from an initial value of 0.44 +/- 0.27 to 4.1 +/- 0.7 units/mg supernatant protein. Addition of LH (NIH-S-18; 2.0 mug/ml) or prostaglandin F2alpha (2.8 X 10(-5) M) to the medium from day 4 to the end of incubation period caused a slight but significant reduction in 20 alpha-SDH activity (20%, P less than 0.05). Supplementation of the medium with ovine prolactin (HIH-P-S11; 10.0 mug/ml) from the time of seeding or from the 2nd to 4th day of culture reduced the activity of 20 alpha-SDH measured on day 12 by 61% (P less than 0.001). This finding suggests that the suppression of 20 alpha-SDH by prolactin, hitherto demonstrated only in vivo results from a direct action of the hormone on the luteal cell.


Androgen regulation of transferase II activity in the mouse kidney. Castration of adult male mice reduced the ability of the transferase factors of the kidney to stimulate amino acid incorporation by polysomes in vitro. The administration of testosterone propionate (TP) to the mice for 11 days greatly increased the activity of the transferase fraction. The changes occurred only in transferase II. The induction of the increase in transferase activity was evident 24 h after the injection of TP and required a lag period of at least 12 h. The concentration of pH 5 enzymes (protein) was slightly decreased by castration and was restored by TP administration. The radioactivity in the hot perchloric acid extract of the protein after amino acid incorporation was increased but the activity of the pH 5 enzyme fraction on amino incorporation was not significantly changed by androgen administration.


Circadian [3H] leucine incorporation into protein of the rat brain: sex differences. The incorporation of 3H from [3H]leucine into protein in various regions of the brain and in the anterior pituitary was measured at 6-h intervals throughout a 24-h period in groups of male and female rats. No rhythm was observed in the male, while there were significant circadian rhythms in certain areas of the female brain, with peaks at 15.00 h. Statistically significant rhythms of the acid-soluble tritium and of the protein concentrations in the individual tissues were also observed, but were unrelated to the rhythms of incorporation of the label into protein.


Protein concentration in immunological preparations and pyrogenic reaction of rabbits. The drop of arterial pressure was found to be caused by proteins injected intravenously into test animals. The drop was related to the protein concentration and the volume of the dose. Low pressure was caused neither by the histamine nor by the protein characterized by its property of liberating histamine. Antihistamines had no influence on quantitative or qualitative depressory reaction, caused by the presence of dissolved proteins. Concentrated protein solutions, when applied intravenously, did not contribute to identifying the pyrogenic reaction, but had a damaging effect on the organism and homeostasis of animals used in the experiments.


Regularities of the frequency of minus temperatures in the rabbit pyrogen test. The author draws the attention on the fact that in the pyrogen test on rabbit lowering of temperature is frequently observed; the drop can reach minus 0.6 degrees C or more. Attempts to explain this phenomenon are presented.


Pyrogen tests of infusions, blood anticoagulant solutions, plastic materials and rubber products. The methods of the pyrogen test in rabbit as adopted by the authors are presented. The test includes positive and negative controls. The conditions of using the same rabbits on two consecutive days are discussed. Methods of sampling of sterile infusions and the preparation for pyrogen test of anticoagulant solutions containing citrate, phosphate and/or edetate ions are presented. The necessity of pyrogen control of distilled water is stressed. Attention is called on the importance of testing for pyrogenicity of the plastic materials and the rubber-wares to be applied during the production of anticoagulant solutions and infusions. A pyrogen test highly sensitive for detecting traces of detergent is applied for washed glassware. It is emphasized that sensitive pyrogen tests are indispensable not only when new derivatives are being introduced, but also during routine control, because occasional changes in the manufacturer's technology may sometimes be demonstrable in this way.


Comparative pyrogen reactivity of rabbit and man to human albumin and immunoglobulin solutions. According to the prescriptions of the Hungarian Pharmacopoeia regarding the pyrogenicity of human albumin and immunoglobulin solutions, the temperature increase should not exceed 0.5 degrees C in rabbits. If the average rise in temperature of rabbits is between 0.6 to 1.1 degrees C, the preparation--in other respects meeting the requirements--can be tested in man (generally 20 persons per preparation). Products causing an average rise in temperature less than 0.6 degrees C and provoking individual rises of temperature below 1 degrees C in man, can be issued. On the basis of these data going back for many years, no reliable correlation could be found between the pyrogen test in rabbits and temperature rise in man. The same preparation which failed to pass the rabbit pyrogen test proved to be apyrogenic in man.


Irreversibility of toxoids. An improved method of testing. Reversion of toxoid to toxin has been described by several authors yet present day regulations of all National Control Authorities have no specific requirements to check for this possibility. Most producers however have included some form of test in their own protocols for release. We have developed a satisfactory test for both diphtheria and tetanus toxoids which involves storing diluted toxoid at 20 degrees C and 34 degrees C for 6 weeks and performing a full toxicity test on the samples. In the case of diphtheria toxoid we have evidence that it may be dangerous to rely on the simpler guinea-pig skin test, since we have shown that several lots, while giving a negative skin reaction, exhibit diphtheria toxicity when given systemically. It is felt that an in vivo conversion to toxin may occur in some lots which would not be detected by the skin test.


The limulus amebocyte lysate test micromethod and application in the control of sera and vaccines. To estimate the amount of endotoxin in sera and vaccines relatively high quantities of limulus lysate are necessary. Because this makes the control rather expensive, a micromethod was developed in which the amount of limulus lysate was reduced fivefold. This method was used to estimate endotoxin in typhoid vaccines. The relation between the reactions in man and the amount of endotoxin in the vaccines was examined. In these experiments it appeared that the limulus amebocyte lysate (LAL) test demonstrates predominantly the endotoxin in solution. The test was also used as a control test for calf sera. The culture quality of these calf sera as estimated by the lymphocyte stimulation test appeared to be influenced by previous bacterial contamination as could be demonstrated with the LAL test.


In vivo and in vitro innocuity tests applied at the National Institute of Haematology and Blood Transfusion. The system of testing for toxocity in the authors' laboratories is presented; it includes toxicity tests in rabbits of the sodium citrate and citric acid ingredients of anticoagulant solutions. The necessity of controlling the biological activity of containers, rubber-ware, accessories, etc. applied in infusion and blood preservation practice is stressed. Innocuity tests are performed in vivo, in the course of the production of the plastic materials to be used, from extracts of the ingredients, of intermediary products and of the finished product. The fish test has proved to be a sensitive method for demonstrating biologically harmful degradation products occasionally appearing in extracts of samples taken during the production and subsequent treatment of plastics. As to in vitro testing, experiences with the 51Cr release test and the control of the haemolyzing effect are presented. Testing of silicon tubes and rubber stoppers used in blood preservation and of infusions and anticoagulant solutions as well as the innocuity tests for anti-D immune globulin and other plasma fractions are presented.


The encephalitogenicity of the Hempt vaccine. The encephalitogenic property of rabies vaccines of brain tissue origin (Hempt vaccine produced in different institutes, Semple vaccine, International Standard of Rabies Vaccine) was studied in animal experiments. Guinea pigs inoculated with the vaccines in complete Freund adjuvant were observed for clinical signs and histopathological changes characteristic of experimental allergic encephalomyelitis. Normal sheep brain suspension was used as control. The minimal encephalitogenic dose of the Hempt vaccine was found about 3,500 times more than that of the control material. Hempt vaccine proved much less reactogenic than the International Standard of Rabies Vaccine and the Semple vaccine. Subsequent lots of Hempt vaccine were slightly different in respect of their encephalitogenic activity.


Studies on the toxicity of a variety of human vaccines as measured by laboratory tests. The results obtained from our laboratory tests for pyrogenic activity on 8 bacterial vaccines are compared with the clinical reactogenicity of the same 8 vaccines in man. It is concluded that, although none of the tests correlate exactly with clinical reactogenicity, the rabbit pyrogenicity test provides the best indication of such activity. The limulus test, although valuable as a means of detecting contaminant endotoxin in pyrogen free preparations appears to be of little value as a gauge of pyrogenicity in bacterial vaccines.


Comparison of the toxic effect on mice of a massive dose of French BCG after low, medium and high dose pretreatment of the French, Glaxo and Japan BCG strains. Numerous laboratory experiments and field trials for tuberculosis prevention have been analyzed. The results concern the comparison of different BCG strains, the correlation between the protection and allergenic property and the degree of residual virulence as an inherited character in producing post vaccination side effects. There are more and more reports about incidence of side effects following BCG immunostimulation. We used the French, Glaxo and International Reference (Japan) strains for this toxicity test because these are the most used strains for vaccine production. The differences obtained in toxic effects of the three tested BCG strains in this experimental model suggest that the investigations should be continued and comparative analyses with histological and immunochemical methods should be extended.


Toxicity testing of peptide for human use. The extent and design of animal tests to obtain information on the possible risks involved with the use of a new peptide in man is still a matter of dispute. It might be relevant to carry out subacute or prolonged toxocity studies in animals also on peptides of human origin to evaluate the possible risk involved if systematically administered peptides reach distant target organs in much higher concentrations than normally, especially peptides whose biological action is close to the site where they are secreted. It seems possible to handle synthetic analogues of naturally occurring peptides similar to any other synthetic compound concerning toxocity testing in animals. Improved methods of peptide synthesis and of chemico-physical characterization may in the future decrease the problem of differences between batches. Today tests should be carried out on hypotensive effect in cats and perhaps also a subacute toxocity study undertaken in animals on each batch prepared by the solid phase technique.


Pertussis vaccine: reactivity in animal and man. Clinical investigation of the reactivity of diphtheria-tetanus-pertussis (DTP) vaccines is a routine control in the GDR. This paper shows the results of the clinical control of 26 batches of DTP vaccine in comparison with animal tests (weight-gain-test, histamine-sensitizing test, dermonecrotic test). The animal tests gave no support to innocuity. The batches were tested on 427 children and passed the requirements for reactivity. After a storage for 2 years the control of 3 different batches was repeated. Two of them showed side reactions in children in spite of the fact that the results of the weight-gain test did not change.


Pertussis vaccine: mouse-weight-gain (toxicity) test. Toxicity tests based on mouse-weight-gain (whether control of pertussis, DPT, cholera or typhoid vaccine) are not satisfactory, particularly since the vaccines may be adsorbed and non-adsorbed preparations. Comparison between results of tests in animals some of which were treated with vaccine and others with saline does not allow to make conclusions as valid as those supplied by a standard control vaccine (adsorbed and not adsorbed) instead of saline.


Comparison of the pyrogen tests in rabbits and with limulus lysate. In the past years an assortment of samples of plasma proteins, enzymes, vaccines and blood substitutes were tested comparatively in rabbits (pyrogen test, European Pharmacopoeia) and with the LAL test (Pyrogent, Byk-Mallinckrodt, Inc.). Specificity and sensitivity were tested with endotoxins and lipid A of gram-negative bacteria. The limulus amebocyte lysate (LAL) test gave similar results or was tenfold more sensitive than the assay in rabbits. More than 300 samples of drugs were examined by both tests. All preparations positive in the rabbit test were positive in the LAL test too. In the testing of plasma proteins the LAL test was more sensitive. The examination of 45 samples of vaccines for pyrogens gave the same result in both assays. Streptokinase does not inhibit the LAL test unspecifically. The LAL test is not an alternative but an additional method in the detection of lipopolysaccharides in drugs.


Comparison of the limulus amebocyte lysate (LAL) test with the rabbit test: false positives and false negatives. Although the test for pyrogens on rabbits, described by the European Pharmacopoeia, is efficient for the detection of clinically important concentrations of pyrogens, it has some practical disadvantages; for instance, it blocks the further use of a rabbit for three weeks once this has been used to test a solution which proved to contain pyrogens. The LAL test for bacterial endotoxin lipopolysaccharides was explored as an alternative method for the detection of pyrogens in infusion fluids and medicinal preparations. False negatives in the LAL test were obtained on some corticosteroid preparations and on some infusion fluids from a batch which had caused febrile reactions in the clinic.


Tests alternative to the rabbit bioassay for pyrogens. The rabbit bioassay is currently the only legally acceptable method to test for pyrogenic contamination of parenteral preparations and medical devices designed to enter the parenterum. However, research efforts have recently made available alternative procedures to test for pyrogens which may have significant value in quality control. Part of the reluctance to accept tests other than the rabbit test may be due to the low visibility of emerging understanding of a significant theory of testing in which it is contended that the parameters of sensitivity and specificity are inversely related and that accuracy is an ideal. The rabbit pyrogen assay will detect 1 to 10 ng of enterobacteriaceal endotoxin (ET). The limulus test will detect 0.01 to 0.1 ng/ml of ET; some of the other tests approach the rabbit assay in sensitivity. Since it is current dogma that pyrogen is equivalent to ET, the basis for the use of ET to standardize pyrogen tests is rationalized. The source of ET in practice is bacterial contamination; therefore, numbers of bacteria that contaminate parenteral preparations can be directly related to potential pyrogenicity. Further, viable counts of bacteria in parenteral preparations, prior to sterilization, is a reliable test for pyrogens. Other tests such as nitroblue tetrazolium reduction and actinomycin-D enhancement of lethality of pyrogen for mice deserve consideration in quality control procedures. The limulus test, the most practical of currently available alternative tests for detection of ET and therefore, of pyrogen has application where the rabbit test cannot be used. Therefore, control personnel must learn of the availability, performance and interpretation of the limulus test. Practical considerations must govern the choice of an alternative test when the use of a test other than the rabbit bioassay is indicated.


Comparability of pyrogen tests.

Comparability of pyrogen tests. The results of the rabbit pyrogen test carried out in different but similarly equipped Institutes on 36 batches of plasma substitute are comparable. If extreme variations occur in the results of the test the cause is not likely to be found in the conditions and execution of the work, as is shown by testing material from the same bottle. When the material is taken from the same bottle the results obtained by the different Institutes are similar. One explanation of the non-comparable results seems to lie in the cleaning of the bottles. It is therefore essential that manufacturers utilize properly washed bottles. Another cause of non-comparable results seems to be linked with the time factor; as it is known from the literature, pyrogen can disappear during storage.


Detection of endotoxin in biological products by the limulus test. The limulus amebocyte lysate (LAL) test is established as a beneficial quality assurance measure for the parenteral drug industry because of its sensitivity, specificity, and simplicity. Limulus amebocyte lysate reacts with various forms of endotoxin to form an opaque gel under acceptable conditions of pH, temperature, and ionic content. Although certain materials and conditions may alter the lysate-endotoxin reaction, the test is not significantly limited by inhibition or non-specific activation. Many U.S. drug firms apply the LAL test generally for monitoring production water and other ingredients, for an in-process control, and as a supplemental end product test for pyrogenic contamination. Specific applications are made for bacterial and viral vaccines, antineoplastic agents, radiopharmaceuticals and drugs designed for intrathecal injection. Efforts to standardize LAL test technique and lysate potency continue.


Studies of the sensitivity and reproducibility of pharmacopoeial pyrogen testing. Rabbits generally numbering 27 or 36 in each experiment were injected in 1970 with doses of the pyrogenic preparation Pyrago (HUMAN) and in 1971-72 and 76 with doses of the International Pyrogen Standard (I.P.St.). The maximum temperature rises were registered, randomized and interpreted according to the requirements of the BP, USP, P. Hung. and P. Nord. 0.29 degrees C mean temperature rises proved to be non-pyrogenic according to BP and P. Hung; whereas according to the USP and P. Nord. in some percents 'to be rejected' qualifications were also established. At more increasing mean temperature rises 0.49 degrees C and 0.58 degrees C, increasing pyrogenic and decreasing 'passable' qualifications were obtained. Beginning with 0.70 degrees C and at 0.86 degrees C and 1.08 degrees C, 'passable' groups disappeared. Exclusion of the rabbits showing low sensitivity (before randomization) barely influenced the results with 3.5 ng kg-1 I.P.St. in the experiment in which the mean temperature rise was 0.49 degrees C. If, however, the mean temperature rise was higher (0.57 degrees C or 0.69 degrees C), such a selection practically resulted in the disappearance of 'passable' qualifications in the triplet groups and a great predominance of 'to be rejected' qualifications in the large groups.


Remarks on some properties and effects of sodium nucleinate. The pyrogen standard of the Pharmacopoeia of the German Democratic Republic--which is recommended in the Compendium Medicamentorum and the Pharmacopoeia of the CMEA countries, consists of a selected batch of sodium nucleinate, the pyrogenicity of which had been demonstrated in very many rabbits. One ampoule of this standard contains 10 mg freeze-dried sodium nucleinate. After inoculation of 0.4 mg per kg of body weight into rabbits an increase in temperature of 0.8 degrees C or more will occur. Westphal assumes that the reason for the pyrogenicity of sodium nucleinate is possibly due to its obligatory contamination with bacterial lipopolysaccharides. Some investigations to find out if the properties and effects of this standard are different from those of isolated bacterial lipopolysaccharides have been performed by the author: adsorption by plastic granulates, inactivation by heating and by incubation with horse serum, temperature changes produced by inoculation into rabbits and results of investigations using the limulus amebocyte lysate test. The results of the investigations did not show significant differences between sodium nucleinate and isolated bacterial lipopolysaccharides. This is in agreement with Westphal's hypothesis mentioned above.


Limulus test, parenteral drugs and biological products: an approach. The authors report their experience with the LAL test for endotoxin assay. They have examined parenteral drugs, blood products and biological substances employed in immunology. The results were compared with rabbit pyrogen tests. All the data suggest that if correctly performed, the LAL test is a simple, rapid and sensitive test. This assay appears to be very useful for materials employed in immunology in order to test the possible contamination with bacterial endotoxin, since the interference between endotoxin and immunologic response is well known.


Early protein malnutrition in the rat: behavioral changes during rehabilitation. Behavioral development was studied in 2 groups of rats during the postweaning period. Pups were selected from litters nursed by dams fed either a standard protein (25% casein by weight) or a low protein diet (12% casein) during lactation. Two pups from each litter were housed together and fed the control diet throughout rehabilitation. Behavioral observations were made by repeated time-lapse photography at 5-day intervals. Differences in home cage behavior were observed at the onset of rehabilitation in the postnatally malnourished rats. Increased feeding behavior was observed during the 1st week of rehabilitation. Locomotor behavior was depressed during the rehabilitation period in the experimental animals. Climbing activity, however, was significantly greater. Self-grooming activity was elevated throughout rehabilitation in post-natally malnourished animals. During the course of rehabilitation, the behavioral differences between the 2 groups gradually disappeared.


Maternal pituitary-adrenal responsiveness as a function of differential treatment of rat pups. The effects of litter separation and pup treatment on the maternal pituitary-adrenal system were investigated in 3 experiments. Lactating females did not show a pituitary-adrenal response to separation from their pups. However, the lactating females showed an increase in plasma corticosterone when their pups had been briefly removed and then returned. If, in addition, the pups were subjected to a noxious stimulus (electric shock) during the 2-min separation, mothers showed a further increase in corticosterone. When pups were returned after 3 hr of separation, mothers again showed a differential pattern of corticoid responsiveness. The magnitude of the mother's pituitary adrenocortical response depended upon the intensity of treatment given to the pups. These data lend physiological support to behavioral studies which have shown that maternal behavior is differentially influenced by pup-produced stimuli.


A critical period for social isolation in the rat. Rats housed in social isolation show heightened levels of object-contact in an open-field and are slower than socially-housed controls to emerge from a small enclosure into an unfamiliar environment. Isolation between 25 and 45 days of age produced an irreversible effect upon object contact but had no lasting effect if between 16 and 25 days or after 45 days. In contrast to object contact, emergence was affected by isolation at any age and the effect was reversed by subsequent social housing. Thus the effects of isolation upon object contact and upon emergence apparently do not depend upon a single underlying variable.


Mirror-image response during the first two years of life. Children aged 1-24 months viewed themselves in planar or distorted mirrors that were either clear or partially occluded. Analysis of videotaped records revealed that interest in viewing one's face did not vary with age among the 1- to 24-month-old subjects. All performed the simple behaviors of observing themselves although the older ones performed more complex (related) behaviors. Neither the flat nor the distorted mirror exposure was preferred suggesting that the infant observes the general configuration of the face for its intrinsic interest. Specific developmental trends were noted in the kind and amount of activity during the 1st 2 years of life culminating in a sequence of behaviors that suggested self-recognition.


Acquisition and extinction of an operant response in differentially reared rats. Rats reared in either socially isolated or control environments were trained to bar press for food on a variable interval schedule of reinforcement, beginning at 125 days of age for 37 consecutive days. Following this phase the subjects were tested for response persistence during an extinction test. Next, the subjects were compared on measures of spontaneous recovery and the rate of response reacquisition when the reinforcement contingencies were once again reinstated. Though no differences were discerned between the groups on measures of acquisition, maintenance, and reacquisition, isolated rats emitted many more responses than the controls during extinction testing.


Early handling influences on behavioral and physiological responses during active avoidance. The effects of early handling on behavioral and physiological responses of the rat during active avoidance learning were investigated. Handled and nonhandled males and females were run in a 2-way shuttlebox task with an unconditioned stimulus of either .5 mA or .8 mA. Animals exposed to .8 mA showed a higher corticosterone response, a shorter response latency, and increased defecation. Early handling did not affect performance of the task, however handling did alter the patterns of the plasma corticosterone response over the course of avoidance training. Handled females showed less elevation of plasma corticosterone than nonhandled females, but all females showed a decrease in corticosterone over the course of avoidance learning. All males showed the drop in corticoids if run at .5 mA shock, but only handled males showed this change at .8 mA shock.


The cardiac self-stimulation hypothesis and the chick's differential attraction toward intermittent sound. Two experiments were conducted to determine whether the chick's attraction toward specific rates of intermittent sound stems from long-term prehatch exposure by the chick to its own fetal heart beat. Contrary to prediction, Experiment I showed stronger attraction toward an intermittent square wave white noise signal matched for pulse rate, overall pulse duration, and mean intensity, than the sound of the actual fetal heart beat from which this attraction is said to derive, Experiment II demonstrated that long-term prehatch exposure to an intermittent auditory signal having a normally non-preferred rate will not establish an attraction for that same rate after hatching unless it contains a frequency component having latent appetitive properties. Alternatives to the cardiac self-stimulation hypothesis are discussed and, in the context of related evidence, the fetus' general capacity for exposure learning is reconsidered.


Development of dustbathing components in bobwhite and Japanese quail. The development of dustbathing components was investigated in Bobwhite and Japanese quail. The incidence and frequencies of components for both species increased with age, but dustbathing occurred earlier in Japanese quail. The dust toss, head rub, and side rub components invariably developed in the same order they occur during an adult bout. The topographies of many components changed systematically with a "rudimentary" form preceding the typical adult form. Large individual differences were observed in the development of the temporal relations among components, but these differences were unrelated to the relative frequencies of components.


Retinopathy and plasma growth hormone levels in idiopathic hemochromatosis with diabetes. In patients with idiopathic hemochromatosis, retinopathy was investigated by ophthalmoscopy and fluorescein angiography and was present in eight out of 23; this prevalence is similar to that reported in patients with diabetes aged between 30 and 60 years at onset of diabetes and with the same duration of the disease; in these eight patients retinopathy was of mild degree, with no impairment of visual acuity, fewer than 10 microaneurysms in each fundus, and no other retinal abnormatic islets directly. These studies confirm that anticholinergic drugs may be useful adjuvants in treating these patients.


Cholinergic blockade in reactive hypoglycemia. The effects of cholinergic blockade on the plasma glucose and insulin responses during oral and intravenous glucose administration were studied. Propantheline (30 mg.) was given by mouth 45 minutes before standard glucose tolerance testing to produce symptomatic chollinergic blockade. In 10 normal subjects a flattening of the over-all plasma glucose response to oral glucose was observed compared with the control test, whereas insulin secretion was not different. In seven patients with repeated episodes of symptomatic reactive hypoglycemia, cholinergic blockade eliminated both symptomatic and chemical hypoglycemia in each, raising the mean nadir glucose from 44 +/- 4 mg./dl. to 84 +/- 8 mg./dl. (p less than 0.01) and significantly reducing insulin secretion. In contrast, following intravenous glucose challenge, cholinergic blockade produced no significant difference in the rate of glucose utilization or insulin secretion in either group. These results are compatible with the hypothesis that excessive vagal stimulation may contribute to the hypoglycemia seen in patients with reactive hypoglycemia but suggest that the predominant effect is on the gastrointestinal tract rather than on pancreatic islets directly. These studies confirm that anticholinergic drugs may be useful adjuvants in treating these patients.


Tolbutamide-induced hemolytic anemia.

Tolbutamide-induced hemolytic anemia. A 67-year-old female diabetic is presented who developed a Coombs'-positive hemolytic anemia after a year of treatment with tolbutamide. An IgG antibody was identified in the patient's serum that caused the agglutination of both the patient's red blood cells and tolbutamide-coated erythrocytes in the absence of complement. Such a reaction did not occur with the patient's erythrocytes when not exposed to tolbutamide. Agglutination of the patient's serum also occurred with erythrocytes treated with other sulfonylureas (chlorpropamide, glibenclamide, carbutamide) but not with phenacetin.


Blood glucose, insulin, and free fatty acid levels during oral glucose tolerance tests in 158 obese children. In 158 obese children, aged from three months to 15 years, blood glucose, immunoreactive insulin, and free fatty acid levels were measured during a standard oral glucose tolerance test carried out prior to treatment. The results were analyzed for the total sample as well as for three age groups: 0-5 years, 6-10 years, and 11-15 years and compared with those of 70 normal-weight children matched for age and sex. Glucose tolerance is normal in the obese children. It is different from the controls only two hours after glucose loading, when a slight but significant elevation is found. The glucose levels at one and two hours are significantly higher in the obese children of group III than in the younger ones. Fasting F.F.A. levels are similar in normal and obese children, but the F.F.A. decrease following glucose absorption is significantly diminished in the obese. The F.F.A. levels of the youngest obese are significantly higher than those of the older ones. A constant and important hyperinsulinism, fasting and postabsorptive, is demonstrated in obese children of all ages, even before five years and at the beginning of obesity. Age- and sex-related differences in insulin secretion are much more marked in the obese than in normal children. The degree of hyperinsulinemia is related to the degree of obesity, but not to its duration. The results suggest that hyperinsulinism is associated with obesity from its onset rather than being a long-term consequence of overweight. However, the origin of hyperinsulinism in obesity and the mechanism of insulin resistance still remain obscure.


Dietary intake and hyperlipidemia in controlled diabetic outpatients. Statistical analyses were performed in order to determine the effects of the control of the diabetes as well as the food intake on serum lipid levels of 73 diabetic outpatients. They had had elevated fasting blood glucose levels, mostly complicated by various grades of hyperlipidemia, before the initiation of treatment. Hyperlipidemia was found to be ameliorated in nearly half of those patients after the start of diabetic treatment. However, the elevation of serum triglyceride persisted in 30 per cent of controlled diabetics, and no differences were found in occurrence of hypertriglyceridemia between diet-treatment group, sulfonylurea group, biguanide group, combined group and insulin group. Estimation of diet intake revealed that the controlled hypertriglyceridemic patients consumed slightly (but significantly) greater amounts of sucrose, alcohol, and total calories than the controlled normotriglyceridemic patients. In addition to such inadequate diet consumptions, the tendency to be overweight and the subtle increment of fasting blood glucose levels were also shown to have contributed to hypertriglyceridemia. It is thus concluded that the lipid disorder in controlled diabetic outpatients is the result of multifactorial influences and that well-conducted diet therapy and stricter regulation of blood glucose are essential in the management of posttreatment hyperlipidemia.


Relation of fasting plasma glucose concentration to plasma insulin and glucagon concentrations. Studies in latent diabetics and women who have produced large-for-dates babies. Mothers who have had gestations diabetes (latent diabetics-LD), as well as those who have produced a large-for-dates baby (LFD) but who were not known to have been diabetic, have raised fasting plasma glucose levels, and these may induce fetal overnutrition. The increased birthweight of babies of obese mothers may also be due to their raised fasting plasma glucose levels. LD and LFD have normal or raised fasting plasma insulin levels even though they have both decreased insulin secretion to small changes in plasma glucose and normal or increased insulin sensitivity. The high fasting plasma glucose probably results from the decreased insulin-secretory response to glucose. Normal subjects have little day-to-day variation of their fasting plasma glucose, whereas subjects with a high fasting plasma glucose have less precise control. Although LD and LFD had abnormal insulin responses, they have normal plasma glucagon concentrations that do not correlate with glucose tolerance or insulin sensitivity. The reported abnormalities of glucagon in diabetes are probably a secondary, not a primary event.


Habitual physical activity and glucose tolerance. Males age 16-64 in a total community. The data being reported are part of an epidemiologic study of health and disease in an entire community, Tecumseh, Michigan. Approximately 1,300 males age 16-65 were classified into three groups on the basis of their habitual leisure and occupational physical activity. A blood sample was drawn one hour after a glucose challenge and analyzed for glucose concentration. A measure of body fatness (sun of four skinfolds) was also available on the men. Glucose concentration of the three physical activity groups was compared by analyses of variance within narrow age group. There was no significant relationship between glucose tolerance and habitual physical activity. There was a slight, but in some age groups significant, relationship between glucose tolerance and body fatness. The population was then divided within each age group into subgroups (thirds) by sum of skinfolds. The analysis was repeated in each age-fatness group. Glucose tolerance was better in the active men but only among the leanest subgroup of subjects.