[Solid phase radioimmunoassay for plasma testosterone using plastic microtiter tray (author's transl)].

In order to simply radioimmunoassay for plasma testosterone and to measure many samples at the same time, a method of solid phase radioimmunoassay utilizing a plastic disposable microtiter tray (DMT) by which chromatography can be omitted was investigated. The antiserum was obtained by immunizing rabbits with testosterone-3 BSA which had been synthesized according to the Erlarnger's method. Plasma samples (male: 0.05ml, female: 0.2 ml) were extracted with 1.0 ml of ether. After freezing the plasma layer in an acetone-dryice bath, the ether phase was transfered to a glass tube and evaporated to dryness. These samples and the dried standard testosterone were dissolved with borate buffer containing 3H-testosterone and transfered to plastic DMT which had been precoated with the diluted antiserum, and incubated for 24 hrs. After removal of the incubated solution, the cups of DMT were cut off and were dissolved with toluene scintillator in counting vials. The radioactivity was counted with a liquid scintillation counter. Other steroids except for 5alpha-dihydrotestosterone (5alpha-DHT) had a low degree of cross reactivity with the antiserum. Five alpha-DHT which could be measured together with testosterone in this assay was not a problem clinically because of its strong androgenic activity. The best standard curve was obtained when the antiserum was diluted to 1:1000. The sensitivity of this assay was 10 pg/tube. The maximal adsorption of antibody to plastic DMT was observed when the pH of antiserum was within the range of 6.5-9.5 and the precoating time was 24 hr at room termperature. The best pH of incubation buffer was 8.0, and the antigen-antibody reaction became a plateau when the incubation exceeded 6 hrs. Water blank in this assay was 4.6 +/- 2.1 pg/tube. The recovery of testosterone (50, 100, 200 pg) added to 0.1 ml female plasma was 99 +/- 6.8%. Coefficients of variation within assay and between assay were below 11.2% and 20.0%, respectively. Correlation between this method and the dextran-coated charcoal method was fairly good (r=0.938). Plasma testosterone levels in 10 normal males and 12 normal females were 616 +/- 202 (mean +/- SD) ng/dl and 66 +/- 29 (mean +/- SD) ng/dl, respectively. The levels were low in patients with hypopituitarism, hypogonadism and acromegaly. They were normal in patients with Cushing's syndrome due to adrenal hyperplasia and adenoma, but they were high in a patient with adrenal carcinoma. In a patient with testicular feminization, the level was 632 ng/dl. This increased after the administration of HCG, and decreased to 127.5 ng/dl after castration. This solid phase radioimmunoassay (using plastic DMT) is economically feasible as well as simple because it is possible to separate the bound hormone from the free hormone of all samples at the same time and there is little restriction in time and temperature. According to the above results, this method is suitable for routine clinical use.


[Stimulatory effect of prostaglandins on biosynthesis of thyroid hormones and plasma TSH levels in the rats (author's transl)].

It is now postulated that prostaglandins (PG) are widely distributed in mammalian tissues and stimulate adenyl cyclase-cyclic AMP system in various organs. The present study was undertaken in an attempt to clarify the effect of PG on pituitary-thyroid axis in vivo. Rats were maintained on a low-iodine diet for 8 days. The animals received graded doses (10-500 mug) of PGE1, PGE2 and PGF2alpha ip daily from day 5 to the day of autopsy. The 4-hr thyroid 131O uptake was consistenly higher in 10 mug PGE1, 10 mug PGE2 and 100 mug PGF2alpha-treated rats than in the controls. Higher T3/T4 ratios were found in the diagested thyroids of PG-tristration. However, a single injection of PG did not affect thyroid 131I uptake in iodine-deficient animals. No change in the thyroid 131I uptake was found in iodine-replete rats chronically treated with PG. The increase in plasma TSH levels during treatment with 30 mug PGE1 and PGE2 was clearly shown. A similar result was also obtained with 100 mug PGF2alpha. The increase in thyroid 131I metabolism produced by the repeated administration of PG in iodine-deficient rats is due presumably to the direct stimulation on the thyorids of rats. In addition, PG elevated plasma TSH levels in such animals, probably mediated through the hypothalamic-pituitary system.


[Comparison of blood glucose and insulin concentrations in man after intravenous injection of 1.0, 0.1 and 0.01 mg of glucagon (author's transl)].

The effect of various doses of glucagon on hyperglycemic action and insulin secretion was studied in 12 healthy volunteers to determine whether 1.0 mg of glucagon, the usual dose, is indeed necessary to obtain an adequate hyperglycemic response. For this purpose 2.0, 1.0, 0.5, 0.1 and 0.01 mg of glucagon (Novo) were used respectively to confirm the existence of a dose response relationship. Of these doses, three different doses were administered to one subject within a week after overnight fasting. Maximum increment of blood glucose was not statistically significant between 1.0 (32.5+/-4.4mg/dl) and 0.01 mg (29.5+/-2.7 mg/dl), averaging about 30 mg/dl. The peak time of blood glucose concentration was 30 min. with 1.0 mg, and 10 min. with 0.01 mg. On the contrary, insulin secretion after glucagon injection decerased as the dose decreased. Maximum blood insulin (IRI) concentration was 69.7+/-17.2muu/ml with 1.0 mg, and 25.3+/-11.3 muu/ml with 0.0l mg (p less than 0.05). The peak time of insulin concentration was 2 min. with 1.0 mg, and 10 min. with 0.01 mg. From these data it is concluded that the dose response relationship after intravenous glucagon injection of between 1.0 mg and 0.01 mg doses exist concerning insulin sercretion, but not concerning hyperglycemic response.


[Studies on the renin-angiotensin system in adrenal regeneration hypertension (author's transl)].

1) To clarify the role of adrenal enucleation on plasma renin activity (PRA), plasma renin substrate (PRS), PRA response to furosemide administration and vacular reaction to renin in adrenal regeneration hypertension (ARH), serial changes of PRA and PRS during adrenal regeneration, PRA response to furosemide administration, and pressor response to exogenous renin in ARH were investigated by comparison with those of intact rats, unilaterally adrenalectomized rats, and unilaterally nephroadrenalectomized rats with contralateral adrenalectomy or with contralateral adrenal exploration (control) on both tap water and high sodium intake. 2) The control rats drinking saline, when compared with intact rats drinking tap water, showed significant decreases in PRA and, concomitantly, significant increases in PRS throughout the experimental period. In the unilaterally nephroadrenalectomized rats drinking saline, two days after adrenal enucleation or adrenalectomy, a significant increase in PRA, with a concomitant decrease in PRS, was observed. Those changes were less pronounced in the adrenal enucleated group than in the adrenalectomized group. Ten days later PRA markedly decreased to the control level in both groups. PRS rose to the control level on the 10th day after adrenal enucleation without increasing further, while that in the adrenalectomized group remained as low as before. 3) No significant differences in any of the experimental groups were found in diuresis, natriuresis, or in changes in body weight and hematocrit during the one and a half hours after furosemide administration performed at the 9th experimental week. The basal PRA was significantly decreased in the other groups with unilateral nephroadrenalectomy and/or a high sodium intake as compared with the unilaterally adrenalectomized rats drinking tap water. The decrease in basal PRA was much more pronounced in the unilaterally nephroadrenalectomized rats drinking saline, with or without adrenal enucleation. After furosemide administration, PRA significantly increased in the unilaterally adrenalectomized rats drinking saline as well as in the unilaterally nephroadrenalectomized rats drinking tap water, with or without adrenal enucleation, while PRA values in three groups were only a half of the unilaterally adrenalectomized rats drinking tap water. An insignificant increase was found in the unilaterally nephroadrenalectomized rats drinking saline, independent of adrenall enucleation. 4) Pressor responses to hog renin in rats with ARH at the 10th postoperative day, and the 4th and 9th postoperative week did not show any significant differences as compared with those of intact rats drinking tap water and unilaterally nephroadrenalectomized rats drinking saline. 5) The effects of adrenal enucleation on PRA, PRS, PRA response to furosemide administration and pressor response to renin in ARH were discussed based on the observed results.


[Psychoendocrinological studies on the pituitary-adrenocortical system in rats. part 1: Relationship between emotional stress (novel stimulus) and plasma corticosterone (author's transl)].

The effect of emotional stress on the pituitary-adrenocortical system in the rat was studied. Hall's open field apparatus was used to give a novel stimulus, a mild and purely not show any significant differences among all the experimental groups. Psychological stimulus produced by a novel environment. Animals were male albino rats of the Wistar-strain, bred in groups and weighing about 250 gm. They were taken out of their home cage and placed gently in the centre of an open field, a novel environment. After a certain period of time, they were returned to the cage and decapitated. Plasma corticosterone was determined by the fluorometric method of van der Vies. The results were as follows: 1. When the animals were exposed to the novel stimulus for 3 min., plasma corticosterone showed a definite increase 15 min. after the end of the stimulus, reached a maximal level at 30 min. and returned to the resting level at 60 min. 2. When the duration of the stimulus, i.e. time spent in the open field, was changed, a 15min.-stay in the field gave the maximal rise of plasma corticosterone and 30, 45, 60min.-stays maintained the plateau level. When the 3 min. open field test was repeated every day, the plasma coritcosterone response was definite in the first trial, but was suppressed at the second and absent at the third and fifth episodes. The results indicate that the acquisition of adaptation to a novel stimulus depends upon the repetition, but not upon the duration of stimulus. 3. Plasma corticosterone responded in the same manner as in the standard open field test, even when olfactory, visual and special factor were changed. This indicates that no special cue (s) is responsible for the adrenocortical response to a novel stimulus. 4. The pretreatment of the animals with tranquilizers such as chloropromazine and diazepam showed no inhibitory effect on the plasma corticosterone response to novel stimulus.


[Psychoendocrinological study on the pituitary-adrenocortical system in rats. Part 2: The relationship of the nature of emotional stress and individual emotionality to plasma corticosterone (author's transl)].

In the previous paper the author described the effect of one kind of emotional stress (novel stimulus) on adrenocortical activity. In this study, two experiments were performed; the first experiment was to determine the relationship between the emotional state and adrenocortical response. The second experiment was to compare the effects of different kinds of emotional stress on corticosterone response. 1. An abnormal emotional state, i.e. a hyperemotional state, was induced by raising animals in a solitary condition for 3 weeks. Both group- and single-housed rats were subjected to repeated 3-min open field tests every day. At the first exposure to the stimulus, both rats responded with an almost identical rise of plasma corticosterone. In contrast with the group-housed animals, in which plasma corticosterone response to the stimulus diminished progressively during the repeated trials, plasma corticosterone in the single-housed rats continued to respond with almost the same magnitude as in the first trial. This indicates a defect in adaptation to a novel stimulus in rats raised alone. 2. Another type of emotional stress (communicated anxiety) was produced by an apparatus, called a communication box, in which test animals can sense the reaction of foot-shocked animals in the neighboring compartments. In contrast with the novel stimulus in the open field, the effect of communicated anxiety on plasma corticosterone did not diminish by repetition of the stimulus. This suggests that adaptation to a communicated anxiety stimulus is difficult to acquire as compared to that of a novel stimulus.


The influence of women's work opportunities on marriage rates.

Several authors have argued that increased work opportunities for women have helped to produce a reduction in the average age at marriage in the United States. This paper tests this proposition on data for the 100 largest SMSA's in 1960. Using ordinary least-squares regression, we find that areas of relatively attractive female employment opportunities had relatively low proportions of women ever married in the age interval 22-24. Other variables significantly related to the proportion married in an SMSA are its sex ratio, percent Catholic, and number of inhabitants. A decline in the sex ratio and improvements in female employment opportunities appear to have been equally influential in producing declines in proportions married between 1960 and 1970.


Insulin-like insulinase-resistant material, distinguishable from normal insulin, in juvenile diabetes.

This study compares some properties of the immunoreactive insulin-like material extracted from the urine of children with overt diabetes with that from normal children. Insulin-like species were fractionated by gel filtration and by isoelectric focusing and were tested for sensitivity to an insulin-specific degradative enzyme. Insulin concentration was measured by radioimmunoassay. The major insulin-like component from the urine of ten normal children and fifteen untreated juvenile diabetics and from the urine of four and the serum of one latent diabetics behaved (on gel filtration) as normal insulin, was sensitive to insulinase, and (in all cases studied) had an identical isoelectric point (resolution 0.1 pH units). A proportion of the immunoreactivity extracted from urine (0-4 per cent from normal children, 5-30 per cent from twelve of the thirteen nonobese untreated diabetic children) eluted from the gel filtration column before insulin. This material from diabetic urine was of two size classes, "proinsulin-like" and "mid-insulin," both resistant to degradation by insulinase. Insulinase-resistant immunoreactivity from one patient was analyzed by isoelectric focusing. Urine samples from two obese children with overt diabetes and four children with latent diabetes contained normal proportions (less than 4 per cent) of immunoreactive species larger than insulin. The possible nature and significance of the present insulinase-resistant species are briefly considered.


Antigenicity of "monocomponent" pork insulin in diabetic subjects.

"Single-peak," "single-component," and "monocomponent" insulins have been produced in an attempt to eliminate insulin antigenicity. Recently "single-peak insulin" has been shown to be antigenic. From animal experiments and preliminary human studies it has been claimed that monocomponent (MC) insulin is nonantigenic or only negligibly so. In this study the antigenicity of MC insulin was determined in two groups of diabetic patients. In group 1, seven patients treated with insulin for the first time were given MC insulin for seven to fifteen months. Four of the seven patients developed significant IgG insulin antibodies after four to ten months. In one patient the IgG insulin antibody concentration was high (8.51 mU./ml.). In two patients, IgG proinsulin-specific antibodies were detected. In group 2, fourteen patients with unstable diabetes, insulin allergy, or resistance were changed from conventional to MC insulin. Treatment with MC insulin did not decrease insulin requirement or improve diabetic control when assayed by the M factor. After seven to eleven months of therapy there was no significant fall in insulin antibodies except in two patients in whom corticosteroids had been administered simultaneously. These results differ significantly from those previously reported and could be interpreted as suggesting that insulin itself is antigenic. When the purity of the MC insulin was determined, significant contaminants could be demonstrated in all of ten separate batches of MC insulin. Gel chromatography, polyacrylamide gel electrophoresis, and proinsulin radioimmunoassay were used to identify the presence of nonconvertible insulin dimer, proinsulin, and monodesamido insulin in antigenically significant concentrations. The generation of IgG insulin antibodies in MC-insulin-treated patients cannot be interpreted as a true indication that insulin itself is antigenic. The problem of insulin antigenicity has not been resolved and will not be until a highly purified insulin is available. Unfortunately, the MC insulins do not meet these requirements.


[Effect of beta-phenylethylamine derivatives on the central nervous system. II. Pharmacological effect of the intracerebral administration of metaraminol on the central nervous system].

Six hours after intracerebral administration, the central effects of metaraminol were examined with the following results. 1) Metaraminol in doses of 40 approximately 160 mug decreased spontaneous motor activity in mice as measured by the photo-cell counters method, the wheel cage method or the open-field test. Ptosis and catalepsy induced at these doses. 2) Metaraminol in doses of 40 approximately 80 mug which influenced the spontaneous motor activity in mice, blocked the avoidance behaviour but not escape. 3) Metaraminol in doses of 1.25 approximately 10 mug which did not influence the spontaneous motor activity in mice, markedly blocked the attaching response induced by the electroshock. 4) Ten min after intraventricular administration, metaraminol in a dose of 500 mug induced the arousal pattern of EEG in sensory cortex of cats and this pattern lasted for 30 min. Ninety min after administration, EEG tracings reverted to the drowsy pattern.


[Assay of cardiotonic steroids based on a 2-compartment model].

In an attempt to develop an assay method for cardiotonic steroids (CS), a two compartment model was applied to the whole dog. As representative CS, g-strophanthin, digoxin and digoxigenin were infused continuously to the animal, and the minimal cardiotonic, irregularity and lethal doses were determined, together with indices K1 and K2, representing the rate of uptake and elimination, respectively, of these compounds into and out of a specific site within the myocardium. The order of cardiotonic potency was: g-strophanthin greater than digoxin greater than digoxigenin. The indices K1 (and K2) were the greatest with digoxigenin )digoxigenin), the second greatest with g-strophanthin (digoxin) and the smallest with digoxin (g-strophanthin). The order of the margin of safety as a ratio between the minimal irregularity and the minimal cardiotonic doses was: digoxin greather than g-strrophanthin greather than lethal and the minimal cardiotonic doses. The cardiotonic, irregularity and lethal doses obtained in the present study coincided well with the values obtained in the canine heart-lung preparations, thus lending further support to the idea that effects of the CS appear when the substances accumulated to a definite amount at a specific site within the myocardium.


[Effect of beta-phenylethylamine derivatives on the central nervous system. III. Motor activity changes in mice due to the intracerebral administration of metaraminol].

Influence of metaraminol on behavioural changes in mice was studied with the following results. 1) When metaraminol was injected i.c., spontaneous motor activity as tested by either the photocell counters method or open-field test markedly increased within 90 min after injection, while on the contrary greatly decreased 3 hr later as compared with control group. Thus, these biphasic effects of metaraminol were clearly established. 2) 30 min after injection, metaraminol in doses of 160 mug did not alter brain catecholamines in mice, however, 6 hr after injection, brain dopamine and norepinephrine markedly decreased. 3) When metaraminol was injected into mice pretreated with reserpine of 6-hydroxydopamine, spontaneous motor activity tested by photo-cell counters method markedly increased and anti-6-hydroxydopamine activity of metaraminol was stronger than anti-reserpine. 4) Metaraminol completely antagonized ptosis and catalepsy induced by rerserpine. 5) Six hr after injection, anti-methamphetamine activity was observed.


[Pharmacological action of cattle prostate extracts (PE). V. Effect on the bladder].

As the extract of cattle prostate (PE) is clinically effective in treating prostatic hypertrophy, a study was carried out on urinary bladders of rat, guinea pig, rabbit and dog as well as on guinea pig ileum. Muscle strip of rat and/or dog bladder contracted with PE with increasing spontaneous movement, and was unaffected by atropine. The isolated ileum of guinea pig also contracted with PE, and the contraction was inhibited by papaverine, but not by atropine. A rise in intravesical pressure was observed with increasing spontaneous movement in guinea pig bladder treated with PE, as well as in rabbit bladder in vitro or in situ. The sphincter vesica of guinea pig was dilated by PE as well as by ACh and methacholine.


[Relation of time of administration of drugs to their acute toxicity to experimental animals].

The circadian rhythms of acute toxicity of N-methyl D-aspartic acid, picrotoxin, pentetrazol, strychnine, chlorpromazine and Na-methylhexabital in dd-mice were investigated. The drugs were injected into mice on the hour at 2, 6, 10, 14, 18 or 22 in one day, after which the cumulative mortalities were calculated for 72 hours. Regarding central stimulants, the mortality of mice injected at 22 (o'clock) was lowest, and when injections were given at 2, 10 or 18 (o'clock), the mortality was higher than at any other time. On the other hand, regarding central depressants, the mortality was lowest at 10, and highest at 14 or at 18 o'clock. Thus, the administration time of central stimulants showing the lowest mortality was shifted about 12 hours in comparison with central depressants. As compared to the central depressants, the mortality rate as the result of central stimulants showed a great contrast when injected at 10 o'clock.


[Adrenoceptors and autonomic nerve control mechanisms in the biliary tract].

Effects of sympathomimetics and sympatholitics were studied on the biliary system of guinea-pigs (in vitro) and rabbits (in vitro and in vivo). Contractions and relaxations produced by sympathomimetic amines are mediated by alpha-excitatory and beta-inhibitoray adrenoceptors in the biliary system (gallbladder, common bile duct and sphinctor of oddi) of guinea-pigs. Sympathomimetic amines caused relaxations in the sphincter of oddi in rabbits (in vitro and in vivo) and responses were antagonized by alpha- and beta-blockade or a combination of both. Sympathomimetic amines caused contraction and relaxation on the common bile duct of rabbits. Transmural stimulation to guinea-pig gallbladder and rabbit common bile duct is mediated by cholinergic and adrenergic nerves, alpha-inhibitory adrenoceptor would thus be involved in the postanglionic cholinergic nerve endings in the gallbladder of guinea-pig. No difference was observed in mechanical responses to transmural stimulation in either the isolated sphincter of oddi or the duodenum of rabbits. It is concluded that the guinea-pig biliary system is functionally independent of the gastro-intestinal system and that the rabbit sphincter of oddi is functionally similar to the duodenal wall.


[Effect of dopamine on perfusion pressure in rat peripheral blood vessels].

Rat mesenteric and hind limb arteries were perfused with the rat's own blood under constant perfusion rate, in situ. The effect of dopamine (DA) administered intraarterially was investigated and compared with effects of norepinephrine (NE), tyramine (Tyr), and phenylephrine (PHE). The magnitude of the vasoconstrictive maximum response of both vasculatures to NE and PHE was larger than those to DA and Tyr, and those to DA were larger than those to Tyr in hind limb, but those to Tyr were more potent than those to DA in the mesenteric artery. The vasoconstriction evoked by celiac ganglionic stimulation was much the same to that evoked by NE and PHE. Under cocaine infusion, the rise in perfusion pressure of both vasculatures evoked by NE and PHE. Under cocaine infusion, the rise in perfusion pressure of both vasculatures evoked by NE was slight, but the duration was markedly potentiated. However, the effect of Tyr was suppressed and that of DA was not changed. In rats on alpha-methyl-rho-tyrosine, the responses of both vasculatures to Tyr were reduced and those to NE and DA were not changed. In the reserpinized rat, the effect of NE was not changed and that of Tyr was reduced, while that of D was more evident in both vasculatures. After phenoxybenzamine treatment, vasoconstrictive effect of DA was reversed in mesenteric and reduced in hind limb artery. Vasodilating effect of DA after phenoxybenzamine, was not changed by treatment with propranolol, atropine, and diphenhydramine but was reduced with reserpine and abolished with haloperidol. These results suggest that the vasoconstrictive effect of DA depends partly on indirect sympathomimetic action, and after phenoxybenzamine, DA acts by vasodilating the DA receptor. Potentiation with reserpine may depend on a post-synaptic mechanism and increase in dopamine-beta-hydroxylase activity.


Blockage of induced pseudopregnancy by electrochemical stimulation of the limbic system.

Pseudopregnancy induced by cervical stimulation was inhibited by acute electrochemical stimulation of the corticomedial amygdala or dorsal hippocampus under sodium pentobarbital anesthesia (40 mg/kg) in adult, cyclic female Sprague-Dawley rats. The degree to which pseudopregnancy was blocked depended on temporal conditions of brain stimulation and sodium pentobarbital administration. Pentobarbital alone had a suppressing effect on the incidence of pseudopregnancy, especially when it preceded cervical stimulation. Limbic stimulation before cervical stimulation had a tendency to potentiate the suppression of pseudopregnancy by pentobarbital. After cervical stimulation, hippocampal stimulation tended to inhibit the development of pseudopregnancy, potentiating the pentobarbital suppression, while amygdala stimulation tended to override the pentobarbital blockage of pseudopregnancy. These findings suggest a negative influence of these two limbic structures and pentobarbital on the secretion of prolactin.


The influence of the nematode Syphacia oblevata on adjuvant arthritis in the rat.

The effect of infestation with the nematode Syphacia oblevata on adjuvant arthritis was studied in the rat. Animals with an established infestation with Syphacia were found to have a reduced incidence of arthritis after injection of Freund's complete adjuvant. Infested animals developing adjuvant arthritis were found to suffer from a less severe form of the disease than animals in which infestation had been eliminated with piperazine before immunization.


Serological studies of Actinomyces israelii by crossed immunoelectrophoresis: taxonomic and diagnostic applications.

Crossed immunoelectrophoresis (CIE) with intermediate gel was applied to the serological analysis of Actinomyces israelii to develop a test with high efficiency in the laboratory diagnosis of human actinomycosis and classification of A. israelii. Recently developed standard antigen-antibody systems for A. israelii by CIE were used as reference. The reference systems were based on standard preparations of cytoplasmic and whole cell-associated antigens of A. israelii and a standard immunoglobulin G pool purified from rabbit antisera to formalin-treated whole cells and cell lysates of A. israelii. The specificity of the standard antigens for A. israelii was evaluated in CIE studies by screening for antibodies to components of the antigens in rabbit antisera raised against related bacteria. The standard system for A. israelii based on cytoplasmic antigens formed species-specific precipitins whereas antisera raised against A. naeslundii and/or Propionibacterium acnes precipitated components of the other standard antigens. As a result of these analyses, the standard system for A. israelii based on 10 cytoplasmic antigens was used as reference for CIE studies to detect humoral antibodies to A. israelii in sera from nine patients with actinomycosis. All the sera from the patients formed at the time of diagnosis one or more precipitins in terms of the 10 reference precipitins. Up to five precipitins were found in single sera. Follow-up studies covering a period of one-half year after treatment showed a gradually decreased precipitin response in the course of time. In control sera from patients with newly diagnosed tuberculosis, nocardiosis, deep Candida infection, and aspergillosis, and in sera from healthy blood donors, no antibodies were detected with specificity for the reference antigens.


Natural antibodies in human cerebrospinal fluid to rabbit erythrocytes.

With an indirect agglutination technique using rabbit antisera to human immunoglobulin, IgG antibodies to rabbit erythrocytes were detected in 211 of 221 unconcentrated cerebrospinal fluids from patients with diseases of the nervous system and controls. The titres varied from 0.25 to 128. The highest titre in the control group was 4, the geometric mean 1.5. The titres were highest in the groups with elevated total protein, for instance inflammatory diseases with mean titre of 9.25. The titre divided by total protein concentration did not vary much in the different groups, but tended to be higher than controls in multiple sclerosis, inflammatory diseases and vascular brain lesions, and lower in the other groups. It did not increase parallel to IgG in multiple sclerosis and inflammatory diseases.


The response of Xeroderma pigmentosum cells and controls to the activated mycotoxins, aflatoxins and sterigmatocystin.

The activation of the mycotoxins aflatoxin B1, G1, B2, G2, aflatoxicol and sterigmatocystin by 9S fraction, microsomal preparation (105,000 times g) and supernatant (105,000 times g) of livers of several species was examined. DNA repair synthesis, chromosome aberrations and clone forming capacity were used as endpoints. Cultured fibroblasts of normal persons and DNA repair deficient Xeroderma pigmentosum patients were employed as test subjects. The activation mixtures significantly increase the chromosome breaking function, lethality and DNA damaging effect (measured as DNA repair synthesis) of aflatoxin B1, G1, aflatoxicol ans sterigmatocystin. The DNA repair-deficient XP cells respond to the activated mycotoxins with a low level of unscheduled 3HTdR incorporation as compared to that of control cells, but show a highly elevated sensitivity to the chromosome-damaging and lethal effect of aflatoxin B1 and sterigmatocystin.


Nerve conduction studies in leprosy.

Motor and sensory nerve conduction studies have been performed on the peripheral nerves in the upper and lower limbs of 30 control subjects, and 36 subjects with leprosy from the Aboriginal population of the Northern Territory of Australia. Impairment of conduction was demonstrated in the vast majority of clinically abnormal nerves, and a large proportion of nerves which appeared clinically to be uninvolved. In a third group of subjects, abnormal conduction was demonstrated in a significant number of nerves which were considered to be clinically enlarged but in whom the diagnosis was initially in doubt. The majority of these patients were subsequently proven to have leprosy. It is concluded that nerve conduction studies are of considerable value in the diagnosis and management of leprosy.


Surgical decompression of posterior tibial neurovascular complex in treatment of certain chronic plantar ulcers and posterior tibial neuritis in leprosy.

Seventy-one cases of posterior tibial neurovascular surgical decompression in leprosy are analyzed and reviewed. Thirteen had chronic refractory posterior tibial neuritis while 58 had chronic nonhealing plantar ulcers. The plantar ulcers were associated with posterior tibial neuritis and/or vascular insufficiency. The clinical and operative findings together with the results are presented and the physiopathology of neurovascular compression is discussed. The operative procedure is described. The presence of pale granulation tissue in a nonhealing ulcer seems to be a characteristic finding in these cases. Neurovascular compression in the tunnel, behind and also below the malleolus, was present in all. In operative procedures, the importance of incising the inferior calcaneal bands is stressed. The results show that the neuritis was cured in all cases, while in 53 of 58 cases the plantar ulcers healed in a short period after the decompression. This stresses the value of this procedure. The prophylactic potential of this procedure needs to be evaluated.


Transmission of leprosy within households.

As part of the leprosy control program, population follow-up studies were carried out during 1962 to 1970 in Gudiyatham Taluk, an administrative unit in South India (population: 400,000). More than 97% of the 23,285 contacts from 5,088 families having a leprosy patient were clinically examined using accepted methodology and confirmed as having or not having leprosy. New cases developing among household contacts of leprosy (secondary attack rates) were determined and studied in relation to characteristics of the index case and that of contacts. The secondary attack rate is defined as the number of new cases discovered in the contacts of leprosy patients per 1,000 person-years, which is equivalent to observing 1,000 persons per year. Nearly two-thirds of all new cases were of the tuberculoid type of leprosy and another one-sixth of indeterminate type. Lepromatous and borderline cases each constituted about ten percent of the total new cases. The total secondary attack rate was 6.8 per 1,000 person-years. Compared to an annual incidence rate of 0.8 per 1,000 in the total population, this incidence rate among the contacts is nearly ten times higher. Such enhanced risks are observed clearly and consistently when studied by the number of patients within a family for both sexes and in various age-groups. The rate for females (6.3 per 1,000 person-year) though less, was not significantly different from that of males (7.1 per 1,000 person-year). The differences observed between males and females for each type of leprosy were also not statistically significant. The risks for children (less than 15 years) are significantly higher than those for adults. Among boys, the maximum risk was observed in the age-group 5-14 years, whereas for females the risk is highest in the age-group 5-9 years, dropping down significantly after that. Furthermore, it was observed that significant differences existed between children and adults only in males but not in females. The secondary attack rates almost doubled when there were multiple index cases in the family. Regardless of the number of index cases, the male-female differences were not statistically significant. Attack rates were significantly enhanced when there was a bacilliferous type of leprosy (lepromatous or borderline) in the family. This was true for the specific attack rates of each type of leprosy too. However, a significantly higher proportion of lepromatous and borderline types is also seen when there is a bacilliferous type of leprosy present. The study reiterates the differences in susceptibility to leprosy among males and females, especially during younger ages. Further immunological studies are necessary to determine the differences in host responses in males and females that produce such a characteristic sex-ratio in prevalence of leprosy. There is still a great need to obtain more data on incidence rates both in general population and among contacts on the basis of prospective observation using acceptable statistical technics in design and analysis.


In vitro effect of dapsone on human chromosomes.

The proportion of accumulated metaphases, as well as the frequency of aneuploidies and structural chromosome aberrations were investigated in DDS-treated and untreated (controls) leukocyte cultures from two samples of healthy adult Caucasoid individuals. In one sample, the sulfone-treated cultures differed from the controls in that they contained 0.4 mug of DDS per ml of tissue culture medium plus phytohemagglutinin. In the other sample, these cultures differed from the controls in that they contained a tenfold higher concentration of DDS. The two concentrations of DDS used have not significantly affected the rate of metaphases, while 0.4 mug/ml DDS did not increase the frequency of chromosome aberrations. The same was not true for the cultures treated with 4 mug/ml DDS in which the proportion of aneuploides and achromatic gaps increased significantly.


Fournier's gangrene of the scrotum.

Gangrene involving the male genitalia is extremely rare. It may occur without any predisposing cause. This necrotic process passes through various stages, and is self-limited. No radical surgical procedure is indicated. A case of typical Fournier's gangrene of the scrotum is presented.


Grudge and the hysteric.

Our understanding of the bizarre sexuality of the hysteric began with Freud's discovery that its roots lay in infantile sexuality, but since then little knowledge has been added. This paper argues that the hysteric in early childhood deals with the failures of good-enough mothering and care by precocious sexual development, which is accompanied by an inability to be nurtured by any loving relationship. Sexual solutions to problems prove, for the hysteric, temporary, and inevitably end in grudge and complaints, as the love-object misreads the hysteric's gestures as expressing sexual wishes and desires instead of as a symbolic body language for care and protection. The author briefly examines the nature of the deprivation in childhood and the reason that the hysteric is such a promising yet recalcitrant patient, and offers a case illustration.


Narcissistic personality disorder in an identical twin.

An identical twin with a narcissistic personality disturbance was analyzed. Clinical and metapsychological issues were discussed. Phenomena of narcissistic pathology and twinship clarify one another. Persistence of the twin as a need-satisfying, mirroring, idealized object interferes with the maternal symbiosis, and the separation-individuation experience. If any object, be it twin, mother, nurse or sibling, retards individuation, there may be instability of the self-representation as well as faulty development of ego and superego structures. Confusing and primitive identifications, as occur in twinship, impair the more stabilizing identifications of the oedipal period, and promote narcissistic character malformation.


Fractionation, isolation and characterization of nonhistone chromosomal protein from calf thymus.

1) A method is described for the separation and fractionation of nonhistone chromosomal proteins from salt-urea dissociated calf thymus chromatin. After precipitating DNA in the dissociated chromatin solution with LaCl3, the chromosomal proteins in the supernatant were fractionated by SP-Sephadex C-25 column chromatography using a combination of NaCl stepwise and linear gradient elutions. Much care was taken to prevent proteolytic degradation of the chromosomal proteins during the preparation. 2) Among the protein fractions separated by this chromatography, twenty subfractions were found to be homogeneous on SDS-polyacrylamide gel electrophoresis. These purified proteins account for about 18% of the whole chromosomal protein. Eleven subfractions of these purified nonhistone proteins had ratios of acidic to basic amino acids above 1.0 and the nine remaining subfractions had ratios below 1.0, corresponding to nonhistone proteins of basic character. 3) The molecular weights of the purified nonhistone proteins ranged from 7,400 to 19,000.


Calcium-dependent interactions of F-actin filaments under the influence of troponin components.

The effect of three components of troponin (TN C, I, T) on the gelation of F-actin was investigated by measuring the increase in viscosity at a very low velocity gradient in a rotating viscometer. TN I or TN T greatly enhanced the generation of F-actin. The effect of TN I-C or T-C complex became Ca-dependent: in the absence of Ca, the complex increased the rate of viscosity rise of F-actin, but in its presence this enhancing effect was almost absent. For these actions, the presence of tropomyosin or heat treatment at 45 degrees was not required. These results can be explained in terms of strengthened interactions of F-actin particles bound with TN T or TN I and the release of TN I-C or TN T-C in the presence of Ca.


Fluorimetric and spectrophotometric studies of DPN-linked isocitrate dehydrogenase from bovine heart. Properties of tyrosyl and tryptophyl residues.

The emission maximum of DPN-linked isocitrate dehydrogenase in pH 7.07 buffer is shifted from 317 to 324 nm and fluorescence intensity is decreased when the excitation wave-length is varied from 270 to 290 nm; in 0.2 M KOH, where the fluorescence of tyrosyl residues is almost completely quenched, a further substantial decline in quantum yield of protein fluorescence and a red shift of the emission peak to 339 nm occur. The latter should be due mainly to tryptophyl residues. The enzyme contains 9.4 tyrosyl residues per subunit of molecular weight 42,000 determined spectrophotometrically (295 nm) at pH 13, in good agreement with a tyrosine content of 9.7 by amino acid analysis. No more than 1.1 tyrosyl residues per subunit can be detected up to pH 10.6 at 7 degrees upon prolonged incubation. The increase in absorption at 295 nm with increasing pH is related to loss of enzyme activity and results in a red shift of the emission maximum, and decreased fluorescence intensity. Treatment of the enzyme in a Li+-containing buffer at pH 7.5 with an excess of N-acetylimidazole results in (a) modification of 1.1 tyrosyl residues per subunit, (b) a 30% decrease in enzyme activity, (c) a 6-nm red shift in emission maximum, and (d) a decrease in fluorescence intensity. Manganous DL-isocitrate (1.06 mM) prevents the acetylation of the enzyme. Deacetylation of the O-acetylated enzyme by hydroxylamine completely restores the enzyme activity and reverses the spectral changes. The acetylation studies indicate that the reactive tyrosyl residue does not participate directly in catalysis but may be involved in maintaining the proper conformation of the active enzyme center. A net of 1 of the 2 tryptophyl residues per subunit is perturbed immediately by a number of solvents. This perturbation is not affected by manganous isocitrate, whereas exposure of tyrosyl residues occurs only with time and is prevented by the substrate. The perturbation of the tryptophyl residue is accompanied by a red shift of the fluorescence emission maximum. The more exposed tryptophyl residue may contribute to the energy transfer from protein to nucleotides since the quenching of protein fluorescence upon binding of DPN+, DPNH, or ADP by enzyme results in a blue shift of the emission maximum. Manganous DL-isocitrate (1.06 mM) quenches protein fluorescence by 16% without a shift in emission peak and does not affect the relative extent of fluorescence quenching induced by the nucleotides.


The action of plasma amine oxidase on beta-haloamines. Evidence for proton abstraction in the oxidative reaction.

The action of plasma amine oxidase upon beta-Br-ethylamine beta-Cl-ethylamine, beta-OH-phenylethylamine, and beta-Cl-phenylethylamine was examined. Beta-Br-ethylamine is a substrate and irreversible inactivator of the enzyme. The enzyme becomes covalently labeled by the inactivator. Approximately 2 mol of inactivator are incorporated per mol of enzyme (MW 170,000). The reduced enzyme is not inactivated. The enzyme catalyzes the elimination of HCl from beta-Cl-phenylethylamine to produce phenylacetaldehyde. The rate of the elimination reaction is comparable to the normal oxidative reaction. We conclude that the occurrence of this elimination reaction establishes the ability of the enzyme to catalyze proton abstraction from C-1 of the substrate and that proton abstraction occurs during the catalytic oxidation normally catalyzed by plasma amine oxidase. Beta-Cl-ethylamine is only oxidized to corresponding aldehyde. Beta-OH-phenylethylamine is neither oxidized, nor does elimination occur. It is a competitive inhibitor in the oxidation of benzylamine and in the elimination of HCl from beta-Cl-phenylethylamine.


Characterization of bovine plasma retinol-binding protein and evidence for lack of binding between it and other bovine plasma proteins.

Bovine retinol-retinol-binding protein (RBP) was isolated from serum as a free, uncomplexed protein under experimental conditions in which human, rabbit, and chicken retinol-RBP are present as tight complexes with prealbumin (thyroxine-binding protein). Purified bovine retinol-RBP formed tight complexes with purified human and chicken prealbumin in physiological ionic strength buffers as judged by gel filtration chromatography, hyperchromic effect on the absorption spectrum of retinol-RBP, and changes in the circular dichroism spectrum. Addition of purified human prealbumin to whole bovine serum shifted the elution position of the specific retinol-RBP fluorescence from a gel filtration column, indicating complex formation in the whole bovine serum. It was concluded from this series of experiments that bovine serum lacks a protein with the binding properties of prealbumin and that bovine retinol-RBP has the normal potential binding to human, chicken, and presumably other prealbumins. Bovine retinol-RBP has a molecular weight, amino acid composition, absorption, and fluorescence spectra which are indistinguishable from that of human retinol-RBP, although the magnitude of the optical rotatory strength of the induced circular dichroism signal at 330 nm was 50% larger in the bovine than in the human material (1.65 and 1.1 Debye-Bohr magnetons, respectively). About 12 liters of bovine and human urine were concentrated by pressure dialysis and a search was made for retinol-RBP using gel filtration and ion exchange chromatography. No retinol-RBP was found in either of these species. This suggested that if, indeed, bovine retinol-RBP is filtered through the kidney's glomeruli due to small molecular size (molecular weight 21,000), there are efficient mechanisms of tubular reabsorption.


Rapid purification of phospholipase A2 from Crotalus adamanteus venom by affinity chromatography.

We have used alkyl ether analogs of ethanolamine and choline phospholipids as ligands to purify phospholipase A2 (EC 3.1.1.4) from Crotalus adamanteus venom by affinity chromatography. One of the affinity columns was prepared with rac-1-(9-carboxy)nonyl-2-hexadecylglycero-3-phosphocholine linked to AH-Sepharose 4B via the carboxyl group. Specific adsorption of phospholipase A2 to this column was achieved in buffer containing Ca2+, and the enzyme was eluted in buffer containing EDTA. The two enzymes from this venom were prepared in good yield (greater than 90%), and were homogeneous as judged by polyacrylamide gel electrophoresis. Retention of phospholipase A2 did not occur when the initial irrigant was devoid of Ca2+. These results support the compulsory ordered mechanism for this enzyme proposed by Wells ((1972), Biochemistry 11, 1030-1041) on the basis of kinetic considerations. The second affinity support was prepared with 1,2-dihexadecyl-sn-glycero-3-phosphoethanolamine attached through the amine moiety to CH-Sepharose 4B. Specific adsorption of phospholipase A2 to this column did not occur. These data indicate that the phospholipid base group must be accessible to the enzyme for optimal binding, and that modifications in the alkyl side chains are more desirable when designing affinity matrices for purification of enzymes involved in phospholipid metabolism.


Studies of the cell surface of Dictyostelium discoideum during differentiation. The binding of 125I-concanavalin A to the cell surface.

125I-concanavalin A (125I-Con A) was found to be equally effective as native Con A in binding to and agglutinating cells of Dictyostelium discoideum, suggesting that iodination of the molecule had no effect on the interaction of the protein with the cell surface. Almost all of the 125I-Con A binding to the cells was inhibited by alpha-methyl glucoside. The binding of 125I-Con A to the cells was extremely rapid, and once bound, the molecule was not readily displaced by prolonged incubation or by the addition of excess native concanavalin A (Con A). In contrast, the 125I-Con A was displaced rapidly from the cell surface by alpha-methyl glucoside. The binding of 125I-Con A to D. discoideum was identical at 22 degrees and 4 degrees, and was unaffected by metabolic inhibitors, suggesting that the protein was not subject to endocytosis. The cell surface Con A binding sites became saturated at high 125I-Con A concentrations. Scatchard plots of the data indicated that growing cells possessed 4 X 10(7) sites/cell, all of equal affinity. Similar plots for "aggregation phase" cells indicated at least two classes of binding sites. A small proportion of the sites had an affinity close to that for the sites on growing cells, but the majority of the sites had a markedly decreased affinity. The total number of binding sites increased only slightly during aggregation to 5.6 X 10(7) sites/cell.


Sialic acids. Enzymatic synthesis of Tay-Sachs ganglioside.

A particulate preparation from embryonic chicken brain catalyzed the transfer of N-acetylgalactosamine from uridine diphospho-N-acetylgalactosamine to the ganglioside GM3 (hematoside, sialyllactosylceramide). The kinetic properties of the transferase were determined. The product was isolated and on the basis of chemical analysis and chromatographic behavior was shown to be Tay-Sachs ganglioside (GM2). The particulate preparation also utilized N-acetyl-D-glucosamine and some of its derivatives as acceptors, but partial heat inactivation and substrate competition experiments indicated that the two classes of acceptors, hematoside and N-acetylglucosamine, were substrates for different N-acetylgalactosaminyltransferases. The enzyme that utilized hematoside showed low but detectable activity with analogues such as lactosylceramide and sialyllactose, but no activity with a wide range of other beta-galactosides and glycosphingolipids. These results are in accord with a proposed pathway for the biosynthesis of the gangliosides and for the patterns of these substances in different cell types and tissues.


The circular dichroism of lysozyme.

The circular dichroism spectra of hen egg white lysozyme, and of lysozyme derivatives in which tryptophan residues 62 or 108, or both, are selectively oxidized, have been measured as a function of pH over the range of 200 to 310 nm. Neither Trp-62 nor Trp-108 is principally responsible for the positive rotational strength in the 280 to 300 nm region. The spectrum in the 200 to 230 nm region is nearly the same in the native protein and in the derivatives, and is little affected by binding of saccharide. These results are used to reinterpret the circular dichroism spectra of the lysozymes and alpha-lactalbumins.


Calcaneo-valgus deformity.

A discussion of the essential deformity in calcaneo-valgus feet develops a theme originally put forward in 1961 on the relapsed club foot (Evans 1961). Whereas in the normal foot the medial and lateral columns are about equal in length, in talipes equino-varus the lateral column is longer and in calcaneo-valgus shorter than the medial column. The suggestion is that in the treatment of both deformities the length of the columns be made equal. A method is described of treating calcaneo-valgus deformity by inserting cortical bone grafts taken from the tibia to elongate the anterior end of the calcaneus.


Sylvatic pulmonary hydatid disease a radiologic survey.

The study assesses the radiologic pattern of endemic sylvatic hydatid disease in Canada. Seventy-five pulmonary hydatid cysts, proved surgically and histologically, were studied from a group comprising fifty-five Indians, five Eskimos and four white patients for the years 1949-1972. Cysts were classified as unruptured or ruptured. Epidemiology, life cycle and pathophysiology are considered in addition to radiology. The majority of cysts appeared to be smaller than the pastoral form found abroad, most were solitary, and all showed characteristic features during growth. The features of cyst rupture are classified and assessed.


Segmental multicystic renal dysplasia.

Two cases of "segmental multicystic renal dysplasia" are presented, both of which showed calcification. Arteriographic findings are described and the feasibility of preoperative diagnosis in such cases suggested.


RNA synthesis in the ultrastructural and biochemical components of the nucleolus of Chinese hamster ovary cells.

A correlated autoradiographic and biochemical study of RNA synthesis in the nucleoli of chinese hamster ovary cells has been made.  Quantitative analysis of the labeling indicates that the fibrillar ribonucleoprotein (RNP) component is labeled faster than 80S RNP and 45S RNA molecules, but approaches simultaneously a steady-state 3H to 14C ratio or grains/mum2 after 30 min of [3H]uridine incorporation.  On the other hand, the 55S RNP, the 36S + 32S RNA, and the granular RNP components have the same kinetic of labeling with [3H]uridine.  These results suggest that the fibrillar and granular RNP components of the nucleolus are the ultrastructural substratum of, respectively, the 80S RNP (45S RNA) and 55S RNP (36S + 32S RNA).  The possibility that precursors to 80S RNP exist also in the fibrillar region of the nucleolus is strongly suggested by the rapid labeling of the fibrils on the autoradiographs.


The induction of DNA synthesis in the chick red cell nucleus in heterokaryons during the first cell cycle after fusion with HeLa cells.

Induction of DNA synthesis in embryonic chick red cells has been examined during the first and second cell cycles after fusion with HeLa cells synchronized in different parts of G1 and S-phase. The data indicate that: (i) the younger the embryonic blood the more rapidly the red cells are induced into DNA synthesis; (ii) the greater the ratio of HeLa to chick nuclei in the heterokaryon, the more rapidly the induction occurs; (iii) DNA synthesis in the chick nucleus can continue after the HeLa nucleus has left S-phase and entered either G2 or mitosis; (iv) the induction potential of late S-phase HeLa is somewhat lower than that of early or mid S-phase cells; (v) less than 10% of the chick DNA is replicated during the first cycle after fusion and only a small proportion (15%) of the chick nuclei approach the 4C value of DNA during the second cycle after fusion; (vi) the newly synthesized DNA is associated either with the condensed regions of the nucleus or with the boundaries between condensed and non-condensed regions; (vii) the chick chromosomes at the first and second mitosis after fusion are in the form of PCC prematurely condensed chromosomes); they are never fully replicated and are often fragmentary; (viii) DNA synthesis in the chick nuclei is accompanied by an influx of protein (both G1 and S-phase protein) from the HeLa component of the heterokaryon.


Electron-microscope observations of mitosis and cytokinesis in multinucleate protoplasts of soybean.

Multinucleate soybean protoplasts produced by spontaneous fusion during enzyme digestion of the cell wall initiated cell division after approximately 40 h in culture. The structure of these protoplasts during mitosis and cytokinesis was studied with both light and electron microscopes. Most nuclei did not fuse but divided synchronously. Interphase nuclei was commonly connected by short narrow nuclear bridges. At prophase and metaphase the nuclei appeared typical of those in most higher plants; technical difficulties prevented an adequate examination of protoplasts at anaphase. Telophase was characterized by cytokinesis involving phragmoplast and cell plate formation; however, complete partitioning of the cytoplasm by cell plants was not observed. Numerous coated vesicles were present near to or continuous with the cell plate and plasmalemma. The presence of a few dividing protoplasts with at least double the normal chromosome number suggests that some nuclear fusion occurred prior to mitosis. Very little cell wall material was detected at the margin of the dividing protoplasts.


TLC, GLC and MS of complex lipid mixtures from uropygial secretions.

A general technique of investigation of uropygial gland secretions is described including sample collection and extraction as well as partition, TLC and column chromatography of the crude lipids. The main components, generally ester waxes, are cleaved by CH3OH/HCl followed by conversion into fractions ready for GLC. Some GLC and MS details are reported.


Diurnal variation of plasma vasopressin in man.

Plasma arginine vasopressin (PAV) concentration was determined by radioimmunoassay during the diurnal cycle in 8 recombent healthy male subjects. Two subjects were studied again 3 weeks later while receiving 1 mycles. In 8 out of 10 cycles, a nocturnal increase in PAV was found; there was a progressive rise during the night in 5 subjects and a peak occurred at 2400 or 3400 h. In 1 subject no variation was detected and in another, the pattern was compleetly different. The mean PAV in the 10 cycles was significantly (P less than 0.001) higher during the night than during the day. Dexamethasone did not modify the pattern of variation, but induced a significant (P less than 0.001) decrease in PAV. Hematocrit remained stable throughout the study as did osmolality, except at 2000 h, when a significant (P less than 0.001) increase (5 mOsm) on average occurred in every subject. Blood sugar, sodium or chloride did not account for the observed rise in osmolality and no simultaneous change in PAV occurred. A rise in PAV explains, to some extent, the known nocturnal decrease in urine output. Diurnal variations in PAV must be taken into account in clinical investigations involving vasopressin.


A case of hypopituitarism with diabetes insipidus and loss of thirst. Role of antidiuretic hormone and angiotensin II in the control of urine flow and osmolality.

A 20-yr-old male was found to have diabetes insipidus is association with panhypopituitarism but without any focal neurological lesion being identified. He was initially treated with steroid supplements, the features of diabetes insipidus being controlled with a thiazide diuretic. Eighteen months later the patient lost thirst sensation and stopped treatment, subsequently being re-admitted with severe dehydration, oliguria and focal neurological signs. Further investigation, including brain biopsy, confirmed the presence of an atypical pinealoma which was considered inoperable. Measurements of plasma antidiuretic hormone (ADH) and angiotensin II (AII) concentrations during the severe dehydration showed very high levels of AII, but inappropriately low plasma ADH levels for the severity of dehydration. We consider that the evidence obtained from this case supports the view that the oliguria with hypertonic urine present during severe dehydration was due to a direct renal action of the very high AII levels, possibly supplemented by the residual ADH secretion.


In vitro analysis of experimental avian viral myocarditis.

The cardiotropism of mumps virus in ovo was reproduced in vitro in a comparative study of the replication of mumps virus in cultivated heart, lung, and liver cells from 11-to 19-day-old chicken embryos. The increased proportion of virus-producing cells in the heart was shown not to be due to greater adsorption of virus to heart cells. Neither interferon nor other cellular inhibitory products were responsible for the lower level of production of virus by lung and liver cells. The proportion of cells producing virus in relation to the amount of virus adsorbed and the proportion of hemadsorbing cells that released infectious virus were greater in heart cells; this finding suggested more efficient viral release in the heart than at the other sites. Inhibition of protein synthesis by puromycin after infection rapidly stimulated the production of mumps virus only in liver cells. Thus a relatively late stage in viral synthesis may be attenuated in the less susceptible liver cells, and events late in viral synthesis may determine organ-related differences in the multiplication of mumps virus in ovo.


His bundle electrogram in trifascicular disease: report of a case studied with His bundle electrograms.

His bundle electrograms were performed on a 75 year old female with trifascicular block and digitalis induced junctional block. The usefulness of this technique in understanding the patients' electrocardiographic abnormalities and the relationship to phase-3 and phse-4 block is discussed.


Routes of nonelectrolyte permeability in gallbladder. Effects of 2,4,6-triaminopyrimidinium (TAP).

The organic cation 2,4,6-triaminopyrimidinium (TAP), which blocks the tight junction channels for cation permeability across gallbladder, also inhibits gallbladder permeability (P) to urea and glycerol without significantly affecting P to Cl(-), sucrose, 1,7-heptanediol, or water (osmotic or diffusional permeabilities). These effects together with the comparisons of P's in frog with P's in rabbit gallbladder suggest that sucrose migrates exclusively through the leakage pathway (through where Cl(-) permeates), and that urea and glycerol permeate in addition through both, the tight junction channels for cations and a polar pathway at the cell membranes. Water and 1,7-heptanediol probably permeate mainly through the epithelial cell membranes.


Studies on infectious pancreatic necrosis virus interactions with RTG-2 and FHM cells: selection of a variant virus-type in FHM cells.

Evidence is presented that adaptation of IPN virus (strain VR 299) to FHM cells entails the selection of a variant virus type that differs significantly from the parental, and most representative, RTG-2 virus type in being able to adsorb efficiently to, and form plaques in, FHM cells. The plaque titre of FHM-non-adapted virus stocks (RTG-2 viruses) was reduced by at least 99-99% in FHM cells, while FHM-adapted virus stocks (FHM viruses) produced plaques at equally high titres in both RTG-2 and FHM cells. FHM viruses and RTG-2 viruses differed also in their behavior in RTG-2 cells in respect to plaque size distribution and growth characteristics, but both virus-types were shown to be morphologically identical, and no significant difference in reactivity against specific antiserum could be detected. Analysis of virus in individual RTG-2 plaque isolates or plaque progeny shows that a mutation of relatively high frequency (10(-4) to 10(-5))robably causes the ability to infect the FHM cells efficiently. Only these mutant virus-types were found in FHM plaque isolates.


High resolution investigations with the scanning and transmission electron microscope of haemadsorption binding sites of mumps virus-infected HeLa cells.

Specific changes at the surface of HeLa cells infected with mumps virus were investigated in parallel with the scanning and transmission electron microscope. The distribution of haemadsorption binding sites and virus-induced antigens at cell surfaces was simultaneously studied by labelling virus-specific antigens with peroxidase-conjugated antibodies after haemadsorption. New information was obtained upon the three-dimensional aspect of the red blood cells, the topographical distribution of their binding sites on the infected cells, and the specific structures at the cell surface which are involved in the process of haemadsorption.


Supersensitivity to d-amphetamine- and apomorphine-induced stereotyped behavior induced by chronic d-amphetamine administration.

Guinea pigs exhibit an increased sensitivity to both d-amphetamine- and apomorphine-induced stereotyped behavior following chronic pretreatment with d-amphetamine. This chronic agonist or "innervation" supersensitivity is believed to be a reflection of an increased sensitivity of dopamine receptor sites within the corpus striatum to dopaminergic agonists. The appearance of dyskinetic movement disorders in humans following the chronic use of levodopa or amphetamine may be a manifestation of similarly increased dopamine receptor site sensitivity within the striatum. It is suggested that the animal model of "innervation" supersensitivity may be useful in the investigation of these human movement disorders.


Multiple cystic and focal encephalomalacia in infancy and childhood with brain stem damage.

Two cases are described in which damage to the brain stem was associated with extensive necrosis of the cerebral hemisphere. In the first case--a monochorionic twin--there was clear evidence that injury of an ischaemic or hypoxic type had occurred during fetal life and some evidence that an inadequate share of the placental circulation was an important aetiological factor. In the second case death occurred 4 yr after an asphyxial episode at birth. The lesions in the hemispheres and brain stem were extensive, although less than in the first example. The lesions are discussed in the context of our knowledge of the anatomy and physiology of the developing nervous system. Although they cannot as yet be fitted into the concepts of "critical periods" and "vulnerable periods" of development, this is perhaps because observations on human cases are scanty in comparison with the extensive animal studies which have been reported. The lesions are contrasted and compared with those seen in animals.


Evaluation of various protein sequences on the nutritional carry-over from gestation to lactation with first-litter sows.

First-litter sows and their progeny were used to evaluate the interaction effects of three gestation crude protein levels (9, 13, or 17%) and two lactation protein levels (12 or 18%) on sow reproduction and litter performance. Diets were formulated to contain the desired protein level by altering the ratio of corn and soybean meal. Daily rations of 1.82 kg were individually fed during gestation and were offered ad libitum to the sows by 7 days postpartum. Litters were equalized to eight pigs each by 1 week of age and held constant to weaning (28 days). Sows fed the 9% protein diet gained less during gestation than those fed the 13 or 17% protein diets. Progeny parturition data were similar for the three gestation protein levels. Sows fed the 18% protein lactation diet and their litters performed similarly in regards to feed intake and progeny performance regardless of previous gestation protein level, while those sows fed the 12% protein lactation diet had decreasing lactation weight losses, but increasing fed intakes and litter gains as the level of gestation protein increased from 9 to 17%. There were significant gestation by lactation protein level sequence interactions on sow feed intakes and litter gains, suggesting a nutritional carry-over effect from gestation to lactation. Sows fed the 17% protein gestation diet had the greatest tissue buffer while those fed the 9% protein gestation diet had the least. These results further demonstrate that dams fed a 9% protein gestation diet perform similarly to those fed higher gestation diets, if the lactation dietary protein level is adequate to meet the lactation amino acid requirements.


Free fatty acids identified as antitryptic factor in soybeans fermented by Rhizopus oligosporus.

The trypsin-inhibitory activity observed in cooked soybeans fermented by Rhizopus oligosporus (fungus used in tempeh fermentation) has been examined. The active compounds have now been isolated by ethanol extraction and thin-layer chromatography and have been identified as free fatty acids by infrared spectroscopy and gas-liquid chromatography. Oleic, lineoleic, and linolenic acids are primarily responsible for the increased trypsin-inhibiting activity of cooked soybeans after fermentation. The free fatty acids are liberated from oil in the soybeans by fungal lipase, and they differ from other reported soybean trypsin inhibitors that are protein in nature. Free fatty acids have been previously reported to inhibit various enzymes, such as glycolytic, glyconeogenic, lipogenic, and also proteolytic. Their effect appears to be a nonspecific type of inhibition. Further studies are required to determine their physiological relevance, if any.


Milk production and nitrogen utilization in response to postruminal infusion of sodium caseinate in lactating cows.

Five lactating, rumen-fistulated Holstein cows were used to obtain additional information concerning the effects of postruminal infusion of sodium caseinate on milk production and amino acid utilization. A 7-day continuous abomasal infusion of approximately 450 g/day of sodium caseinate was preceded and followed by 7-day infusions of an isonitrogenous-isocaloric solution of glucose, monosodium glutamate, and urea. Total collections of milk, urine, and feces were obtained during the last 5 days of each infusion period. On the last day of each period, arterial and mammary venous blood samples were obtained for analysis of plasma free amino acids. During infusion of sodium caseinate, milk production, milk protein (N times 6.38) production, and efficiency of nitrogen utilization for milk crude protein production were increased. Arterial plasma concentrations of free histidine, isoleucine, phenylalanine, valine, and total essential amino acids were elevated above control levels during infusion of sodium caseinate, while ornithine and tryrosine were decreased. Calculation of the relative concentration of essential amino acids in arterial plasma and in milk protein indicated that methionine and lysine were least abundant in plasma relative to their requirement for milk protein synthesis. A high precentage extraction from arterial plasma by the mammary gland also suggested that methionine and lysine may have been the essential amino acids in most critical supply for milk protein synthesis. Calculation of uptake to output ratios of individual plasma amino acids by the mammary gland suggested that significant quantities of extracted arginine, isoleucine, leucine, threonine, and valine were utilized in pathways other than direct incorporation into milk protein.


Quantitation of the biosynthesis of immunoglobulin in peripheral blood lymphocytes of normal and immunodeficient patients.

A solid phase radioimmunoassay was applied to the study of biosynthesis of immunoglobulins in lymphocytes cultured from peripheral blood of normal and immunodeficient patients. Total immunoglobulin and IgG were readily detected in lymphocyte culture fluids in studies of all normal individuals (age range 28 weeks, gestation to adulthood). In contrast, synthesis of immunoglobulins was absent or markedly reduced in cultures from patients with humoral immunodeficiencies. Inasmuch as this method requires relatively small amounts of blood, it may be of particular value in the diagnosis of humoral immunodeficiencies in infants and young children.


Radiation effects on rat testes. V. Studies on lysosomal enzymes (acid phosphatase and acid DNAse) and their physiological significance following partial body gamma-irradiation.

Acid phosphatase is present in the nucleus and cytoplasm of cells in the seminiferous tubules and the interstitium of rat testes. The effect of irradiation on acid phosphatase is dependent on the environmental temperature and the dose of irradiation. It appears that initial rise in the enzyme at a low radiation dose and a high environmental temperature or at a high dose and low temperature is associated with a lysosomal breakdown of the germinal cells of the testes. A decrease in acid phosphatase in the advanced stages of radiation injury is a secondary radiation effect which may lead to decreased metabolic synthesis of phosphate esters owing to the unavailability of orthophosphate in the testicular tubules. The reduced acid phosphatase activity can be detected in the seminiferous tubules, suggesting that the enzyme activity is related to the state of the germ cell population. An initial increase in acid phosphatase is matched by an initial rise in acid DNAse within hours of irradiation, further suggesting that there is radiation interaction with the cells of the germinal epithelium. The enhanced activity of DNAse following a 2nd week of irradiation at 2000 R confirms the phagocytic activity of the non-germinal cells.


The penetrability of rabbit ova treated with enzymes or anti-progesterone antibody: a probe into the nature of a mammalian fertilizin.

The acrosome reaction of rabbit spermatozoa, an essential prerequiste for penetration of the zona, occurs usually in the vicinity of the egg, suggesting that the rabbit may produce a factor akin to the 'fertilizin' of some invertebrates. Specific inactivation of such a factor should render eggs impenetrable and possibly point to the nature of a 'fertilizin' in mammals. Rabbit eggs with granulosa cells removed were treated for different periods with trypsin, chymotrypsin, neuraminidase or anti-progesterone antiserum, and then transferred alone, or together with control eggs (one group labelled with fluorescein isothiocyanate), to the oviducts of inseminated recipients. Three hours later the eggs were recovered and the experimental and control groups were compared for penetration of the vitellus and for numbers of spermatozoa within the perivitelline space or in the zona pellucida. None of these treatments affected the penetrability of the zona pellucida significantly since the number of spermatozoa within treated eggs in any one experiment was always comparable to that of untreated eggs exposed to the same fertilization environment. If there is a specific substance emanating from or present on the surface of the rabbit egg which induces the acrosome reaction, its activity seems unaffected by trypsin or chymotrypsin; the charged radicals of N-acetyl neuraminic acid or local concentrations of progesterone do not appear to be involved.


Synthesis and some pharmacological properties of (3-beta-(2-thienyl)-L-alanine)-8-lysine-vasopressin.

[3-beta-(2-Thienyl)-L-alanine]-8-lysine-vasopressin was synthesized by solution techniques. The partially protected heptapeptide Boc-Cys(Ec)-Tyr-Thi-Gln-Asn-Cys(Ec)-Pro (1) was synthesized in a stepwise manner using the active ester method or the dicyclohexylcarbodiimide (DCC) coupling technique mediated by 1-hydroxybenzotriazole (HBt). The protected nonapeptide amide Boc-Cys(Ec)-Tyr-Thi-Gin-Asn-Cys(Ec)-Pro-Lys(Coc)-Gly-NH2 (2) was prepared by coupling 1 with Lys(Coc)-Gly-NH2 using DCC-HBt. From 2, [3-thienylalanine]-8-lysine-vasopressin was obtained by removing the Boc-protecting groups with trifluoroacetic acid and ethylcarbamoyl (Ec) protecting groups in refluxing liquid NH3 followed by oxidative cyclization in H2O-MeOH using ICH2CH2I. Purification was effected by partition chromatography followed by gel filtration. The highly purified product possesses activities in the oxytocic, avian vasodepressor, rat pressor, and antidiuretic assays of 19.0 +/- 0.5, 87 +/- 4, 243 +/- 5, and 332 +/- 32 units/mg, respectively. Thus [3-thienylalanine]-8-lysine-vasopressin has higher oxytocic, avian vasodepressor, and antidiuretic potencies than does 8-lysine-vasopressin, whereas its pressor potency is about the same as or slightly lower than that of 8-lysine-vasopressin.


Synthesis and neuroleptic activity of isomeric thieno(1,4)benzothiazines.

To investigate the influence of electronic properties of the tricyclic thiazine system on neuroleptic activity, a series of the isomeric N-dimethylaminopropylthienobenzothiazines was synthesized. All compounds were screened for neuroleptic activity in mice and rats. For the active compounds lowest active doses in the antiamphetamine test were determined. Activity appeared to be dependent on the mode of annelation of the thiophene molecule: compunds bearing the same substituent and side chain with the thiophene molecule in 2,3 and 3,4 annelation were active, while those compounds with a 3,2 annelation seemed to be devoid of activity at the given dose.


4-Trifluoromethylimidazoles and 5-(4-pyridyl)-1,2,4-triazoles, new classes of xanthine oxidase inhibitors.

The syntheses of a number of 2-substituted 4-trifluoromethylimidazoles and 3-substituted 5-(4-pyridyl)-1,2,4-triazoles are described. The trifluoromethylimidazoles were prepared from 3,3-dibromo-1,1,1-trifluoroacetone after hydrolysis with aqueous sodium acetate solution and condensation with an aldehyde in the presence of ammonia. Basic hydrolysis of the trifluoromethyl group was found to provide a facile method for the synthesis of imidazole-4-carboxylic acids. In the imidazole series a 2-aryl substituent and a free imino group were required for xanthine oxidase inhibitory activity. The triazoles were obtained through the reaction of an aroylhydrazine and an imino ether followed by thermal ring closure of the intermediate acylamidrazone. As in the imidazole series, a free imino group is an absolute requirement for in vitro activity. Additional structure-activity relationships of these compounds are presented.


Gene flow versus selection pressure and ancestral differentiation in the composition of species: analysis of population variation of Juniperus ashei Buch. using terpenoid data.

Twenty-four populations of Juniperus ashei were sampled throughout the range of this taxon and the terpenoids of the foliage were analyzed by gas/liquid chromatography.  Population differentiation was investigated by analysis of variance and numerical taxonomy.  Three south Texas and one Mexican population clustered together with the rest of the taxon appearing fairly uniform.  No evidence was found of hybridization or introgression with other taxa.  Disjunct populations in Oklahoma and the Ozarks, which have been genetically isolated from the central population for thousands of years, showed no signs of differentiation nor genetic drift.  The present pattern of distribution probably dates from the Pleistocene.  The south Texas and Mexican populations appear to be the primitive elements of the species.  Populational differences have apparently been maintained in adjacent populations in spite of seemingly large gene flow and conversely, chemical uniformity is being maintained in many disjunct populations where there is little or no gene flow.


Effect of lifetime exposure to aflatoxin b1 in rats.

Aflatoxin B1 was fed at 2 ppm in the diet to a group of pregnant F344 rats from the time of conception; it was then fed to their offspring until death. This diet was also given to another group of rats 6-7 weeks old for comparison. The survival time of male rats was significantly shorter than that of the female rats of both groups. However, the survival times of rats of the same sex in both groups did not differ significantly. The major causes of death were hepatic neoplasms with matastases, although some early deaths occurred before neoplasms developed. Most deaths were from a malignant hemorrhagic liver tumor, histologically diagnosed as a hemangiosarcoma, which caused rupture and hemorrhage into the peritoneal cavity or metastases to the lungs. These hemangiosarcomas were readily transplantable and did not produce alpha-fetoprotein. Ultrastructurally, they were composed of poorly differentiated cells resembling endothelial cells. Nodules of hyperplasia induced by aflatoxin B1 sometimes grew large (greater than 1.5 cm), and 2 were transplanted. Approximately 20% of the rats had colon tumors; a few rats had tumors of the kidney, oral cavity, and hematopoietic system.


In vitro stimulation of human breast tissue by human prolactin.

Normal human breast tissue was cultured with defined media plus hormones. The epithelium survived at least 4 days in culture but did not grow in the absence of hormones. Both insulin and human prolactin stimulated growth, but ovine prolactin did not.


Virus replication and cell modifications in organ cultures of tumor tissue from chickens with Marek's disease.

Organ cultures of lymphomas from chickens infected with Marek's disease (MD) virus were examined by electron microscopy at various periods after explantation. Non-enveloped herpes-type virions  were observed in explants 18 days after the culture was initiated. Cytolytic virus was replicated often in lymphoblastoid cells and occasionally in other cell types (permissive and nonpermissive transformed). The permissive cells were numerous, but nonpermissive transformed cells were also present as indicated by the longevity of some cultures. The presence of both permissive and transformed cell types in the organ cultures and reasons for their sequential appearance were discussed. The results unequivocally demonstrated that lymphoid cells of tumors induced by MD virus were sites of virus replication under proper culture conditions.


RNA:DNA ratios in a developing fibrosarcoma and its lung metastases in C3H mice.

The quantitative ratios of RNA:DNA were followed in a developing transplanted fibrosarcoma in C3H mice and in its lung metastases. There was a significant increase in these ratios in developing tumors originating from cell suspensions (P smaller than 0.001) and a single implanted piece (P smaller than 0.05). No significant change was demonstrated in developing fibrosarcoma originating from two pieces of this tumor (P greater than 0.05) which were implanted simultaneously. When comparing the ratios of RNA and DNA of developing lung metastases to the primary tumors, we found a significantly higher ratio in the metastases (P smaller than 0.001). No significant changes in RNA:DNA ratios were demonstrated in normal proliferating tissues either in physiologic hyperplasia or embryo tissue culture (P greater than 0.05).


Sindbis virus glycoproteins: effect of the host cell on the oligosaccharides.

Sindbis virus was grown in four different host cells and the carbohydrate portions of the glycoproteins were analyzed. Sindbis virus grown in BHK-21 cells has more sialic acid and galactose than Sindbis virus grown in chicken embryo cells. In other respects the carbohydrates from virus grown in these two hosts are very similar. Sindbis virus grown either in chick cells transformed by Rous sarcoma virus or in BHK cells transformed by polyoma virus was also examined. In comparisons of virus from normal and transformed cells, differences in the amount of sialic acid were observed; but otherwise the carbohydrate structures appeared basically similar. The growth conditions used for the host cell also affected the degree of completion of the carbohydrate chains of the viral glycoproteins.


Biochemical studies on bovine adenovirus type 3. I. Purification and properties.

Bovine adenovirus type 3 (BAV3) was purified and its properties were studied. On productive infection of CKT1 cells (a cell line derived from calf kidney) with BAV3, it was observed that viral DNA synthesis was initiated after about 24 h and its rate was maximal after about 40 h. Maturation of the virus occurred several hours after this. Purified BAV3 was separated into four discrete bands by CsCl density gradient centrifugation (complete, incomplete, empty, and degraded viruses). The complete BAV3 was similar in size and structure to human and avian adenoviruses. Polyacrylamide gel electrophoresis showed that the complete BAV3 virion contained at least 10 polypeptides. The total structural proteins of the virion had a similar amino acid composition to those of human adenoviruses. DNA of the complete virus was a linear duplex and its contour length was 12.3 +/- 0.9 mum. The So20,w value of the DNA was 32.9S and its buoyant density in CsCl was 1.717 g/ml. There was about 25% homology between the DNAs of BAV3 and human adenovirus type 5 by filter hybridization. It was also noted that BAV3 produced incomplete virus. The incomplete virus was similar in morphology to the complete virus and contained almost all the structural polypeptides of the latter, but lacked infectivity. However, its DNA had a deletion(s) (13%) which seemed to locate near a terminal.


Biochemical studies on bovine adenovirus type 3. II. Incomplete virus.

Incomplete virus of oncogenic bovine adenovirus type 3 (BAV3) was highly purified and its biological activity was studied. The production of incomplete virus was found to increase with a high multiplicity of infection and with a large amount of arginine in the growth medium. On infection of contact-inhibited mouse cells, incomplete virus induced cellular DNA synthesis and focus formation. Moreover, this virus was oncogenic to newborn hamsters. On infection of calf kidney cells, a permissive cell line, viral early and late RNA, viral DNA, and almost all the viral late proteins were produced, but no mature progeny virus was detected. It is, therefore, suggested that incomplete virus of BAV3 may be unable to synthesize a protein(s) (perhaps a kind of maturation protein[s]) essential for assembly of viral macromolecules for maturation.


Low frequency of (5'-3') -C-G- connection in 70S RNA from simian sarcoma virus.

The frequency of oligonucleotides obtained from simian sarcoma virus RNA by digestion with ribonuclease T1 was compared with the frequency expected of an RNA molecule in which nucleotides are arranged in random distribution. Oligonucleotides containing C-residue attached to 3'-Gp were found significantly less in simian sarcoma virus 70S RNA than expected by random distribution.


Skin testing in genitourinary carcinoma: 2-year followup.

Compound 2,4 dinitrofluorobenzene was used to sensitize 115 patients with genitourinary cancer to test their ability to be immunized with a newly encountered antigen. In general, patients in whom a cutaneous hypersensitivity response did not develop were more likely to have advanced stage tumors than patients whose immunocompetence was intact. A 2 to 3-year followup showed that patients with impaired cell-mediated immunity were more likely to undergo disease advancement than patients of similar stage whose immunity was normal. Unfortunately, these results cannot be extrapolated for application to a given patient.


Duplex kidneys: a correlation of renal dysplasia with position of the ureteral orifice.

A study was made of 51 duplex kidneys, ureters and bladders to assess the types of abnormalities seen in duplex kidneys. The abnormalities affected ectopic and orthotopic segments equally. The occurrence of renal abnormality was closely correlated to the abnormal locations of the segment's ureteral orifice. When the orifice was displaced either cranially (orifice positions B, C and D) or caudally (orifices G and H) the orthotopic and ectopic segments, respectively, showed more severe hypoplasia and dysplasia. Faulty embryogenesis is proposed to explain these findings and the importance of endoscopic positioning of the ureteral orifice is emphasized in determining whether heminephrectomy will be necessary.


An unusual pattern of the mitral echocardiogram observed in cases of congestive cardiomyopathy and other myocardial diseases.

(1) Narrowness of the early-diastolic and presystolic peaks on the echo curve of the anterior mitral leaflet and an abnormal step formation at the middle or higher level on the descending limb of the presystolic peak were observed in 7 cases with congestive cardiomyopathy or myocardial fibrosis, 1 case with myocardial infarction, and 1 case with cardiomyopathy due to progressive muscular dystrophy. (2) This abnormal pattern is considered to be closely related to the myocardial condition in the above-mentioned diseases. (3) Probable mechanisms for this pattern formation are considered as follows: (i) a sudden reduction of distensibility of the left ventricle after filling of the ventricle over a certain limit near full-filling, and impaired ventricular contractility, (ii) restriction in the mobility of the mitral valve and its chardae due to thier inability to adapt themselves to a developed dilatation of the left ventricle. (4) A similar presystolic step formation on the echo curve of the anterior mitral leaflet was observed in 3 of 25 cases of hypertrophic cardiomyopathy of Goodwin's sense. Differences between the myocardial state in congestive cardiomyopathy or in similar myocardial diseases and that in hypertrophic cardiomyopathy were also discussed.


Cell-mediated immunity to Dirofilaria immitis.

Cell-mediated immunity to Dirofilaria immitis (DI) in guinea pigs was confirmed by the migration inhibition test (MIT), the blast transformation test (BTT), the delayed skin reaction, and the skin reaction by passive transfer with sensitized peritoneal exudate (PE) cells. All migration inhibition (MI) positive cases were always associated with positive skin reactions and two cases showed positive skin reactions without MI. The cellular antibody confirmed by MIT first appeared on the 4th day after single sensitization, but DNA synthesis in splenic lymphocytes had already started on the 3rd day in the absence of delayed skin reaction and MI. Then, the role of this cellular antibody in the immune mechanism against DI infection was investigated by the in vitro and in vivo cytotoxicity test using microfilariae (Mf) of this species as a target. The cytotoxic activity significantly increased in the sensitized splenic and PE cells, and in vivo normal PE cells implanted into sensitized animals.


[A trial for electromyography of the detrusor muscle (author's transl)].

The author developed a new electromyography of the detrusor muscle which is applicable also for clinical examinations and obtained the following results: 1. In the normal bladder no action potential appeared when the organ was not full yet. 2. When the intravesical pressure was raised by introducing saline into the bladder, spikes appeared. 3. Spikes of high frequency appeared with the onset of voiding. 4. In the normal bladder no action potential appeared by administration of 5 to 7 mg mecholyl. 5. From some muscles of the dome of the bladder was obtained spontaneous spike potential consisting of spike burst of about 10 spikes. In an animal experiment using adult dogs the spikes of the simple type appeared. The discharged interval was about 5 seconds. 6. In the bladder with damaged nerve, spikes appeared during veridical quiescence. 7. In the bladder with damaged peripheral nerve, spikes of high frequency appeared by administration of 5 to 7 mg mecholyl.


[Electromyogram of the alimentary tract by chronically implanted bipolar electrodes (author's transl)].

For the long term observation of the electrical activity of the canine alimentary tract, a bipolar silver electrode was devised in our laboratory. The electrode has been demonstrated to be satisfactory in sensing the electrical activity of the alimentary tract. The electromyogram of the esophagus was consisted of very fast spike burst lasting for about 3 to 5 seconds in swallowing actions. The stomach showed a cyclically recurring basic electrical rhythm (BER), which was propagated caudally, and spike burst which was related to the contractile activity of the gastric smooth muscle. The duodenum and small bowel also showed both BER and spike burst in their electrical activity. Thus the electromyograms obtained by our electrode have shown their similarity to those by monopolar electrodes. Furthermore, our electrode has proved its durability, functioning for more than 5 months. Details of the construction of our electrode were described.


[The effects of flavoxate hydrochloride on the gastrointestinal motility (author's transl)].

The experiments were performed to study the effects of Flavoxate hydrochloride on the gastrointestinal motility and its underlying mechanism. Gastrointestinal motility was inhibited at first and then accelerated markedly after the intravenous injection of Flavoxate hydrochloride (10 mg/kg) in the experiments of anesthetized dogs. For analyzing the underlying mechanism of this response the following experiments were executed. 1) The pendular movements of isolated ileum of rabbits were accelerated with administration of low concentration of Flavoxate hydrochloride less than 10(5) g/ml, while these were reduced with administration of high concentration more than 10(4) g/ml. Because the tetrodotoxin (2.5 times 10(7) g/ml) has no essential effect on these responses, it could be postulated that Flavoxate hydrochloride has direct effect on intestinal smooth muscle itself. 2) Flavoxate hydrochloride (10 mg/kg) induced the differentiated regional response of the sympathetic outflow, that is, the parallel decrease of cutaneous and cardiac sympathetic activities, and the converse increase of splanchnic activity. Therefore, Flavoxate hydrochloride is thought to have an effect on the integrating mechanism of the sympathetic nervous system. This view was also supported by the experiments with neuromuscular preparation of lobster (Panulirus japonicus). It is concluded from these results, that the change of gastrointestinal motility induced by intravenous injection of Flavoxate hydrochloride (10 mg/kg) is influenced both by its effect on the gastrointestinal smooth muscle itself and by its effect via the autonomic nervous system.


[Monitoring heart frequency (beat to beat) and excursions of the thoracic wall--cardiorespirography--during exchange transfusion in newborn (author's transl)].

Heart rate (calculated beat to beat) and excursions of the thoracic wall were registered continuously during exchange transfusions in the newborn period (N equals 29). Short time changes of heart rate ranging from 5 to 10 (N equals 11) and from 11 to 25 beats per minute (N equals 11) were usually present before exchange transfusion. In two cases "silent" (i.e. short time changes of heart rate lower than 5 bpm) heart rate pattern was registered before and in another two cases appeared during the exchange transfusions. This pattern is of prognostic value, 3 of these 4 children showed severe complications during the therapeutic procedure. Of particular interest is, that the "silent" pattern could be detected before other clinical signs of shock appeared. Monitoring heart frequency (beat to beat) therefore gives the possibility to start the therapy of complications early.


[Significance of the 2-deoxy-d-glucose-test on the differentiation of hypoglycemia in childhood (author's transl)].

The 2-deoxy-D-glucose-test (2-DG-test) was applied to 4 children with idiopathic hypoglycemia along with some other tolerance tests. In comparison with the measurement of urinary catecholamine excretion during insulin-induced hypoglycemia, the 2-DG-test seems to be more useful because it is easier to apply and better tolerable. With regard to treatment one should differentiate between an isolated type of adrenal medullary unresponsiveness and a ketotic hypoglycemia with impaired regulation of epinephrine secretion.


[Virologic and serologic findings in an epidemic of abacterial meningitis of unknown origin (author's transl)].

In the summer of 1972 there occurred an epidemic of abacterial meningitis in Bayreuth and its surroundings. We received material of 52 ill children, aged 2 months to 14 years. A virus could be isolated in 10 (5 Coxsackie A4, 2 ECHO 6, 1 Coxsackie B 5 and twice enterovirus which would not be typed). Only in another 13 children could we make unequivocal diagnoses (7 Coxsackie A4, 2 Coxsackie B5, one each Coxsackie A 2, ECHO 3, Mumps, RRS-virus). With the other 29 patients a safe laboratory diagnosis could not be made, because there were either antibody titers against several virus from the same blood specimen (10) or nothing abnormal could be found (19). Therefore, since in a third of those examined no laboratory diagnostic basis for the cause of the epidemic could be found, we felt that we had not established the cause. The large number of positive findings we believe to be due to more or less "normal" epidemiologic processes in this child-population during the summer-fall period. The obvious limits of serologic diagnosis are discussed.


[Genetic studies in hypoplastic left heart syndrome (author's transl)].

The families of the 18 patients with hypoplastic left heart syndrome (HLH), who died in the Universitätskinderklinik Erlangen between 1967 and January 1974, have been investigated. There was no consanguinity of the parents. Only one sib has died of a cardiac failure which possibly could have been HLH. Therefore it could be excluded that HLH is an autosomal-recessive disorder. The overall incidence of cardiac failure in the sibs of patients was 3. Most of the patients with HLH have been born in June-August and December-January. Environmental factors seem to be of importance in the genesis of HLH. Our results suggest multifactorial inheritance of the hypoplastic left heart syndrome.


[Quantitative changes of serum immunglobulins during infectous diseases in childhood (author's transl)].

Immunelektrophoreses and quantitative determinations of serum immunglobulins were performed for 298 children with infections of the upper and lower respiratory tract, chronic and recurrent infections, pyelonephritis and Salmonella infections. Minor changes were seen for IgG, 80% of the patients had levels within the normal range. Many patients, however had increased levels of IgA and IgM, while a decrease of these immunglobulins below the normal range were rarely detected. Children with upper respiratory tract infections had increased IgA in 28 per cent and increased IgM in 44 per cent, those with bronchitis in 21 per cent an IgA and in 45 an IgM increase. Most frequently immunglobulin elevations were seen in patients with pneumonia: IgA was in 50% and IgM in 67% increased above the normal range. Patients with recurrent infections had an IgA elevation in 34% and an IgM increase in 33%. 35% of children with pyelonephritis had an IgA and IgM increase. Children suffering from Salmonella infections had an increased IgA in 29 and IgM in 67%. The result of other authors and of factors leading to an elevation of serum immunglobulins are discussed.