Diurnal variations in plasma corticosterone and growth hormone as corrlelated with regional variations in norepinephrine, dopamine and serotonin content of rat brain.

Statistically significant diurnal variations in plasma growth hormone (GH) were found to occur in handled male rats. Peak GH values (at miday) appeared to be inversely correlated with the diurnal peak of plasma corticosterone(CS) which occurred after the onset of darkness. Brain amines were examined in the following regions: cortex, striatum, septum, amygdala, pons, midbrain, and hypothalamus. Statistically significant diurnal cycles of serotonin (5-HT) concentration were found in the amygdala and midbrain, significant diurnal alterations in norepinephrine (NE) levels. Ultradian dopamine (DA) cycles were significant in all regions examined. Plasma GH changes were found to be directly correlated with midbrain and amygdala 5-HT levels and with DA levels of all areas except the amygdala, Plasma CS  was found to be inversely correlated with striatal  and cortical DA and with 5-HT levels of amygdala.


Effects of pinealectomy and constant light on the estrous cycles of female rats.

Adult Charles River female rats were pinealectomized, sham-operated or left intact, and half of each group was placed in 12L:12D while the other half we exposed to constant light (LL). Daily vaginal smears permitted the calculation, at weekly intervals, of the percent of rats in each group in LL showing 5 or more consecutive days of vaginal cornification. After 11 weeks in LL, those rats were returned to LD for 6 weeks and then put back into LL until the termination of the experiment 3 weeks later.


Ballistic and corrective movements on an aiming task. Intention tremor and parkinsonian movement disorders compared.

Six patients with Parkinson's disease, six patients with essential or intention tremor, and nine controls were tested on a step-tracking task using a joystick control and oscilloscope display. Tremor subjects resembled controls in making an initial ballistic movement followed by (defective) corrections, and took longer than the controls to reach the target with small amplitude jumps, but not with larger ones. The reverse was true for parkinsonian subjects, who acquired the target with slow corrective movements only. This suggests that two kinds of movement available to normal people are selectively impaired in these disorders; ballistic movements in Parkinson's disease and small amplitude corrective movements in the other disorders.


Epidemiologic study of migraine symptoms in young women.

Information about headache was collected from a nonclinical sample of 451 women, aged 15 to 44, in 12 major U.S. cities. Questions were asked in regard to the presence in the past year of headache, and of the following characteristic symptoms of migraine: unilateral location, throbbing quality, visual aura, vomiting, and severity sufficient to affect daily activities. Twenty-three percent of the women had headaches with two or more of these characteristics. The frequency of such symptoms was significantly greater in women who smoked or formerly had smoked cigarettes, in women with lower incomes and poor education, and in women with a history of hypertension, stomach ulcer, fainting, and a variety of emotional complaints. The frequency of reported symptoms of migraine did not vary significantly according to age, race, marital status, use of oral contraceptives, or number of living children. These findings do not support the commonly held clinical impression that migraine is uncommon among blacks or among the poorly educated.


Unilateral megalencephaly: hamartoma or neoplasm?

The clinical and pathologic findings in three cases of unilateral megalencephaly, showing a spectrum of mild to severe involvement, are presented. The first case demonstrated an increase in neuronal size and mild gliosis and is contrasted with the second and third cases, which showed a progressive enlargement and bizarre appearance of neurons associated with an increase in number and size of astrocytes. The relationship of these cases to developmental and neoplastic disease in the brain is discussed.


Adolescent familial cramps.

Two adolescent brothers were studied because of cramps in the legs following strenuous exercise. The symptoms were maximal at adolescence and later subsided. Serum creatine phosphokinase elevations were significant. Examinations of muscle biopsy material revealed changes compatible with a myopathy. The family consists of seven children. All have eventually developed elevations of serum creatine phosphokinase, which appear to reach the highest point during adolescence. This familial disorder may represent a new myopathy with maximal penetrance at adolescence.


Clinical ictal patterns in epileptic patients with occipital electroencephalographic foci.

Seizure patterns and other clinical features were analyzed in 55 epileptic patients with electrographic evidence of exclusive or predominant occipital involvement. Few statistically significant differences in clinical or ictal patterns were found between subjects with purely focal occipital involvement and those with temporal and temporo-parietal spread or minor additional independent foci. On the other hand, cases with bilateral synchronous occipital spike activity appeared to reflect a different type of epileptic disorder. Clinical pleomorphism was more apparent than is commonly conceived; thus, although the incidence of visual auras was relatively high (47 percent), epigastric, psychic, somatic, and other sensory phenomena were not infrequently encountered. Ictal motor patterns were most commonly (53 percent) nonfocal or absent, but partial or focal motor attacks and psychomotor seizures were amply represented. Ictal features with their localizing and lateralizing reliability were also analyzed and discussed in relation to those found in a comparison study of seizures of fronto-centro-parietal origin.


Dystrophia myotonica in a 47, XXY male.

Classical clinical manifestations of dystrophia myotonica were apparently unaltered by aneuploidy in a 47, XXY male. His extremely eunuchoid habitus may represent a contribution by the point mutation to his Klinefelter's syndrome. Association of the two disorders may be due to factors other than chance. The danger of overlooking Klinefelter's syndrome in males with dystrophia myotonica can be avoided by cytologic studies.


Perivascular anoxia-ischemia lesions in the human brain.

The clinical and pathologic data are presented on three patients who had clinical anoxia-ischemia in combination with serum hyperosmolality. The pathologic lesions consisted of perivascular necrosis, predominantly involving small vessels in the cerebral cortex. No intraluminal thromboembolic material was found in any of the vessels. The lesions are reminiscent of "no-reflow" lesions seen in experimental animals rendered ischemic.


Brain H-3-catecholamine metabolism in experimental cerebral ischemia.

Unilateral ligation of a common carotid artery in gerbils causes a major depletion of brain dopamine, which is most marked in brain regions known to receive dopaminergic projections. To determine whether this depletion reflects release of stored dopamine, a radioactive label (H-3-dopamine) was introduced into brain dopamine pools 4 hours prior to ligation. Twenty-four hours later, brain H-3-catecholamines were profoundly depressed ipsilateral to the lesion among animals exhibiting clinical signs of stroke. Within brain regions known to receive dopaminergic projections, common carotid ligation also was associated with a selective decrease in the concentration of H-3-deaminated metabolites. These data suggest that cerebral ischemia is associated with release of catecholamines, as well as with impaired oxidative metabolism of catecholamines.


Low intensity argon laser coagulation in central serous retinopathy (csr).

Mechanisms of light coagulation effects in RPE and detached retina are discussed. 25 cases of CSR are presented in which the leaking point in the RPE was coagulated with an argon laser coagulator, the exposure parameters being set to avoid a whitening of the retina. The results suggest that the retinal whitening and consequently retinal damage does not constitute a criterium for coagulation effectiveness in CSR.


An evoked electromyographic test for peripheral facial palsy.

The purpose of the present study was to determine the degree of nerve degeneration in patients with facial palsy from the number of spikes in M-wave which were induced from the facial muscle by means of a bipolar concentric needle electrode when the facial nerve was stimulated. Ten normal cases between 14 and 72 years of age and 46 cases with facial palsy between 12 and 73 years of age were examined. Histopathological findings of ten denervated nerves were studied to determine to what extent this method may reflect the state of nerve degeneration.


Experimental investigation on lateral inhibition by means of stapedius reflex threshold.

In 15 normal-hearing persons a number of stapedius reflex responses 10 dB above individual reflex threshold has been measured and recorded. In an experimental condition the same stimulus tone has been used and the responses recorded. During the experiment two tones have been continuously presented - one of lower and one of higher frequency with amplitudes lying 10-15 dB underneath the tone evoking the stimulus. In an experimental condition an attenuation of the reflex response resulted. The normal reflex decay, first of all in higher frequencies has been controlled during experiment. The results indicate a similarity to those found under adequate conditions in other perceptive organs, these being interpreted as lateral inhibition.


Aspirin ototoxicity in the guinea pig.

Aspirin ototoxicity has been studied on guinea pigs by shiver-audiometry and histological investigation of the cochlear duct. One dose of 350 mg/kg has provoked, after 7 h, a mean hearing loss of 18-24 dB at 0.25-8 kHz, followed by complete recovery in 3 days. The difference between the administration of 50 mg/kg/day and 350 mg/kg/day consists of a wide extension of the frequencies involved, and in about 10 dB a greater hearing loss. In both cases, no appreciable recovery of hearing was observed after 22 days (the histological investigations were negative). The biochemical pathogenesis of aspirin ototoxicity is discussed and periodical audiometric controls before and during salicylate treatment are recommended.


Extrusion of transtympanic ventilating tubes, relative to the site of insertion.

The results are presented of a study, designed to examine the relation between the site of insertion of transtympanic ventilating tubes into the tympanic membrane and the period, elapsing before extrusion takes place. The results suggest that tubes inserted into the anterior part of the tympanic membrane remain in place for a longer period than tubes inserted posteriorly.


Urethral pressure measurement after division of the external sphincter.

The completeness and permanence of 'division of the external sphincter' is assessed in a group of 30 spinal injury patients. The urethral pressure profile technique is effective in estimating the completeness of the operation. Both in the short-term and the long-term (four month to 16 years) the results of the operation are excellent with no mortality.


The autonomic nervous system and the bladder during spinal shock--an experimental study.

During experimental spinal shock, the urethral muscle responded to drugs earlier than the detrusor muscle. Stimulation with parasympathomimetic drugs caused some rise in intravesical pressure, but an unexpected and much greater rise of intraurethral pressure. Bladder evacuation was achieved with the Credé manoeuvre during spinal shock with the combined effect of alpha adrenergic blockade and beta adrenergic stimulation.


Ontogeny of L-glutamic acid decarboxylase and gamma-aminobutyric acid concentration in human kidney.

GAD activity and GABA, the product of GAD action on L-glutamate, are both prominent in mature human renal cortex. GAD activity is low in fetal kidney but rises several fold preterm to establish the characteristic post-term specific activity. The ontogeny of the initial step in the GABA pathway parallels the need for kidney to accommodate acid-base regulation after birth. PLP coenzyme is required for GAD holoenzyme integrity. Fetal renal GAD was frequently undersaturated with PLP in our series of observations, raising the suggestion that maternal vitamin B6 nutrition is not always adequate.


Pulmonary valvular dysfunction in children with ventriculovenous shunts for hydrocephalus: a previously unreported complication.

A hitherto unreported cardiovascular complication of ventriculovenous shunts for hydrocephalus, involving the pulmonary valve apparatus, is described in two boys. This complication was the result of catheter migration with its proximal end interfering with pulmonary valve function. In one case the embolized catheter was removed by thoracotomy resulting in disappearance of the cardiac findings, whereas in the other case the catheter was in situ because of its peripheral position in the lung and the relatively insignificant hemodynamic disturbances of the pulmonary valve. The pertinent literature on the cardiovascular complications of ventriculovenous shunts for hydrocephalus is also reviewed.


Effect of local cooling on sweating rate and cold sensation.

Subjects resting in a 39 degrees C environment were stimulated in different skin regions with a water cooled thermode. This local cooling produced decreases in sweating rate measured at the thigh and increases in magnitude estimates of the cold sensation. The are of cold stimulation varied from 111 cm2 to 384 cm2. Sensitivity coefficients of the changes in sweating rate and magnitude estimate were corrected for differences in size of the area of stimulation and change in skin temperature and were normalized to the responses of the chest. The normalized coefficients showed the following relative sensitivities for changes in sweat rate and magnitude estimate respectively: forehead 3.3, 2.2; back 1.2, 1.4; lower leg 1.1, 0.9; chest 1.0, 1.0; thigh 0.9, 1.0; abdomen 0.8, 0.8. Varying the area stimulated from 122 cm2 to 384 cm2 produced greater changes in the sweating response than in the magnitude estimate. Rate of skin cooling during the period of stimulation had more effect on the sweating response than on the magnitude estimate. We conclude that cooling different body regions produces generally equivalent changes in the sweat rate and sensation, with the forehead showing a much greater sensitivity per unit area and temperature decrease than other areas.


Spike-free activation mechanism in smooth muscle of guinea pig stomach.

Electrical and mechanical activity of circular muscle strips of guinea-pigs stomach, taken from the distal corpus/proximal antrum region, were recorded. Spontaneous activity consisting of phasic contractions combined with bursts of spike potentials was suppressed by verapamil (5-10-6 - 2-10-5 mol/l). Under these conditions acetylcholine produced a spike-free tonic activation. Under normal conditions phasic contractions were superimposed on this tonic activation. The acetylcholine-induced activation, therefore, consists of two different components, one of which can be selectively blocked with verapamil. Both components disappear quickly in calcium-free solution. It can be concluded that two different calcium activation systems are responsible for the two components of activation. In comparative studies with taenia coli preparations a comparable spike-free tonic activation was not found.


Abnormal relationship between sodium excretion and hypertension in spontaneously hypertensive rats.

Inulin clearance, single nephron glomerular filtration rate (SNGFR), Na and K excretion were studied following an acute saline infusion in spontaneously hypertensive (Okamoto strain) rats (SH). 1. Hypertonic saline load: experiments were performed in adult and young SH rats. As compared to control normotensive Wistar rats (NI), the sodium excretion rose much less following the load in SH. During the 75 min following the beginning of the load, adult SH excreted 15 degrees (NT: 58%) and young SH 9%(NT = 38%) of the sodium load (P less than 0.01 in both cases). Cin were similar in SH and NT during control period, and a similar increase was observed following the load. The superficial to juxtamedullary SNGFR ratio was 0.80 plus or minus 0.05 in 5 non diuretic adult SH, a value not different from that found in normal rats, and 0.87 plus or minus 0.07 in 3 salt loaded SH, indicating that no significant intrarenal GFR redistribution occurs in SH following an acute hypertonic saline load. 2. Following an isotonic saline load, the results were not different: the percentage of the sodium load excreted was 44 plus or minus 5% and 10 plus or minus 2% in NT and SH adult rats respectively. The possibility that the observed defect in sodium excretion of SH rats involves distal tubular function is suggested by the pattern of evolution in Na and K excretion.


Calcium conductance and tension in mammalian ventricular muscle.

Voltage, membrane current and contraction were simultaneously measured in voltage clamp experiments (single sucrose gap) on cat ventricular trabeculae. The pulse programs allowed the determination of the potential dependence of the steady state activation and inactivation as well as the restoration of the calcium-carrying system (slow inward current). 1. The steady state activation variable (d infinity) rose in a sigmoid manner from -50 mV (d infinity nearly 0) to 0 mV (d infinity nearly 1). The experimental values can be described by the function 1/1 + exp [(Vh-V)/S] where half activation (Vh) = -22.5mV and S = 7.6 mV. 2. The steady state inactivation variable (f infinity) declined from 1 at -60mV to 0 at 10mV. The best fit curve is nearly a mirror image of the activation curve with Vh = -28 mV and s = -8.3 mV. 3. The voltage dependence of the (normalized) peak tension was well described by the steady state conductance variables except at potentials positive to +20mV. A "steady state" tension (superimposed on "tonic tension") was found in the potential range where a steady state conductance is predicted by the curves describing steady state activation and inactivation. 5. Following inactivation, the time courses of restoration of the calcium-carrying system and tension were identical. Time courses were exponential with tau = 118 msec at -80 mV, 144 msec at -60 mV, and 198 msec at -40 mV. 6. Two possible models of excitation-contraction coupling in line with the present results are discussed.


Synthesis and spectroscopic properties of two classes of 5,6-dihydrothymidine derivatives. Action on the Ehrlich's ascites cells thymidine kinase.

Trans (+) and (-) 6-alkoxy-5-bromo-5,6 dihydrothymidine and trans (+) and (-) 6-alkoyloxy-5-bromo-5,6-dihydrothymidine compounds have been prepared. The synthesis of these substances (alkoxy : methoxy, ethoxy, butyloxy and isoamyloxy and alkoyloxy : acetoxy and bensovloxy) is described. Diastereoisomers of all products have been isolated by thin layer chromatography and their spectroscopic properties (IR, UV, NMR, mass spectrometry) studied. These compounds have been shown to be competitive inhibitors of Ehrlich's ascites cells thymidine kinase with respect ot thymidine.


Quantitation of Semlike Forest virus RNAs in infected cells using 32-P equilibrium labelling.

In vitro cultured BHK and HeLa cells were labelled for several cell division cycles with 32-P-phosphate until they were equilibrated with radiophosphorus. After infection with Semliki forest virus (or mock-infection) these cells were analyzed for viral and ribosomal RNA by sucrose gradient centrifugation. From their radioactivities the mass of each RNA species was calculated. It was found that the BHK and HeLa cells contained on average 11.0 plus or minus 3.1 pg and 6.3 plus or minus 1.9 pg of ribosomal RNA (28 S + 18 S) respectively per cell. At the end of the viral growth cycle, i.e. at 8 h post infection the average mass of viral genome produced per cell was 1.0 -1.9 pg and 0.3 - 0.5 pg in BHK and HeLa cells respectively, of which only 1/10 to 1/20 was released as mature virus particles. The amount of the second major virus specific messenger, the 26 S RNA, was estimated from its ratio to the viral genome after labelling with 3-H-uridine in the presence of actinomycin D. These two viral RNAs were found to be present in roughly equimolar amounts.


Stereochemical aspects of the interaction between steroidal diamines and DNA.

A complete series of stereoisomeric quaternised diaminoandrostanes, differing in their stereochemistry at the 3,5 and 17 positions, has been examined for effects on the thermal denaturation of calf thymus DNA and for the ability to remove and reverse the supercoiling of closed circular duplex PM2 DNA. In both types of test the eight isomers rank in the same order of effectiveness. The preferred stereochemistry for the quaternary substituents at positions 3 and 17 is beta; of these the orientation of the 17- substituent is more critical. Folding of the steroid skeleton between the A and B rings, as in 5beta-androstanes, diminishes effectiveness but does not necessarily abolish the effect on supercoiling. The over-riding importance of the C-D ring end of the steroid nucleus bearing a 17beta-amino substituent is confirmed by a comparison of the effects of five mon-amino androstanes. Relative helix=unwinding angles per bound steroidal diamine molecule have been determined for four of the isomers; for three 17beta compounds the estimated values are similar to that previously reported for irehdiamine A. For a fourth isomer the angle is 0.22 times that of ethidium, the lowest yet determined for any DNA-binding drug. The results lend further support to the argument that intercalation can be reled out, and alternative models for the binding mechanism are discussed.


Poly (U, s 4-U) as a synthetic messenger RNA for polyphenylalanine synthesis in a cell-free system derivem derived from rat liver.

Two different U and S4-U containing polymers with U:s4U ratio of 3:1 and 5:1 were synthesized. Their activity as messenger RNA was studied in an amino acid incorporation cell-free system from rat liver. It was shown that both copolymers can stimulate the incorporation of phenylalanine into oligophynylalamyl-tRNA.


The effect of methionine and lysine levels on the arginine requirement of the chick.

Two breeds of commercial broiler chicks were used to investigate possible breed differences and to determine the effect of methionine and lysine on arginine requirements.  Graded levels of arginine were added to a glucose-casein diet with or without added methionine and to a corn-soybean meal diet with and without added lysine and/or methionine.  The arginine requirement of chicks receiving the glucose-casein diet with and without supplemental methionine was found to be 1.46 per cent and 1.55 per cent of the diet, respectively.  No breed differences were found.  When arginine was added to a corn-soy diet containing 1.53 per cent arginine, with or without supplemental methionine, no response was obtained indicating that this level of arginine was adequate.  When this diet was supplemented with lysine to bring it up to the lysine level of the casein diet, a growth depression occurred which was overcome by the addition of 0.20 and 0.25 per cent arginine, respectively, in the absence and presence of supplemental methione.  These levels of arginine exceeded the requirements determined for chicks fed the glucose-casein diet.  In chicks fed the glucose-casein diet, muscle creatine increased with each level of added arginine with or without supplemental methionine.  Creatinine excretion also increased with each level of added arginine in the absence of supplementary methionine but when methionine was added creatine excretion reached a plateau at the level of arginine which satisfied the chick's growth requirement.


Relative utilization of serine and glycine by chicks.

Studies were conducted on the relative utilizaiton of glycine and serine by chicks fed basal crystalline amino acid diets devoid of these amino acids.  The crystalline amino acid mixture was fed at one and three times the requirement levels, thereby stimulating uric acid synthesis at differing rates.  In addition, 5 per cent L-glutamine replaced L-glutamic acid on an isonitrogenous basis in three diets containing normal levels of amino acids in the second study.  Chicks fed diets devoid of glycine and serine grew less rapidly and less efficiently than chicks fed diets containing either serine or glycine plus serine.  These decreases were roughly the same whether the diet contained normal or high levels of amino acids.  Serine was as efficient as glycine in supporting chick growth and feed efficiency regardless of whether diets containing normal or high levels of amino acids were fed.  Chicks fed diets containing high levels of amino acids grew approximately 81 per cent as rapidly, but 24 per cent more efficiently, than chicks fed normal levels of amino acids, and excreted approximately twice the amount of uric acid per gram of nitrogen consumed.  In spite of increased uric acid excretion by chicks fed the high amino acid diets, the dietary void in glycine and serine was no more detrimental to chick growth or feed efficiency than that noted when normal levels of amino acids were fed.  Feeding 5 per cent L-glutamine rather than L-glutamic acid in the diet containing normal levels of amino acids had little effect on weight gain, feed efficiency or uric acid excretion.  The absence of cystine from the amino acid mixture used in the third study did not have a marked influence on the relative utilization of glycine and serine by the chick.


Influence of dietary selenium on tissue selenium levels in turkeys.

To study the influence of dietary supplements of selenium on tissue selenium levels, duplicate groups of 20 Broad Breasted White turkeys were fed practical diets alone or supplemented with 0.1 and 0.2 p.p.m. Se as Na2SeO3.  The concentration of naturally occurring selenium in the basal starting and finishing rations was 0.20 and 0.13 p.p.m. respectively.  Selenium analyses were performed on samples of blood, liver, breast muscle and leg muscle obtained at 14 and 20 weeks of age.  Supplemental selenium did not change the level of selenium in any of the tissues studied, nor did it affect weight gains.  The concentration of selenium in blood and muscle was approximately 0.2 p.p.m. and ranged from 0.6 to 0.7 p.p.m. in liver.  The results indicate that supplementing diets containing marginal levels of Se with 0.2 p.p.m. Se as Na2SeO3 does not lead to accumulation of Se in the tissues.


Amino acid bioavailability: a comparative evaluation of several assay techniques.

A fish meal and a soybean meal sample were bioassayed for available amino acid content according to five different published techniques. The techniques consisted of: 1) a two week chick growth assay (GA) for available lysine; 2) a digestibility assay performed by total fecal collection (TFC); 3) a digestibility assay based on the analysis of ileal contents IA); 4) a procedure based on partial fecal collection and analysis of a dietary marker using gnotobiotic chicks (GF); and 5) procedure 4) using conventional chicks (CONV). The GA gave high lysine availability estimates for both proteins, but values for fish meal were significantly reduced when the level of dietary incorporation was increased. The TFC assay proved difficult in collecting adequate sample and, as a consequence, abnormally high availability values were obtained. Modification of this collection procedure resulted in values comparable to the IA which gave the lowest average values of all the assays (90.5 per cent for fish meal and 91.1 per cent for soybean meal). Mean availabilities calculated from GF and CONV assay were significantly higher than the modified TFC and IA techniques. Significant differences were obtained in several individual amino acids when the GF and CONV assay were compared. A growth assay, using diets formulated on the results of these assays, showed that the available amino acid profile obtained from the CONV assay resulted in significantly improved weight gains and feed conversions as compared to a similar diet based on the chemical analysis of amino acids alone. All diets utilizing available amino acid data resulted in improved feed conversions.


[Primary disaggregational thrombocytopathies].

The authors present the results of clinico-laboratory examination of 30 patients with a congenital (hereditary) hemorrhagic tendency associated with an isolated disturbance of the adhesive-aggregation platelet function. Several variants of disaggregational thrombocytopathies were distinguished. Characteristics of hemorrhagic phenomena in the patients is given; a tendency to hemorrhages in them was more frequently detected with the aid of Borchgrevink's test they in determination of bleeding time after Duke.


An angiographic study of the pre- and postoperative hemodynamics in patients with side-to-side portacaval shunts.

Hepatic panangiography was performed in 17 patients before and after side-to-side portacaval shunts. Preoperatively, a direct correlation existed between the diameters of the hepatic arteries and the severity of the patient's cirrhosis. Postoperatively, the splenic artery and vein and the hepatic artery dilated further in most patients, reflecting increased blood flow. The greatest increase occured in those patients with less severe cirrhosis. The liver shrank postoperatively in most patients. The corrected sinusoidal pressure dropped significantly in all patients. A unique pattern of sinusoidal filling at wedged hepatic venography was observed postoperatively in many of the patients.


Action of 7-oxa-13-prostynoic acid on rat uterine contractility and sensitivity to PGF-2 alpha and angiotensin II.

The action of 7-oxa-13-prostynoic acid on spontaneous contractions and on PGF-2alpha and angiotensin II elicited contractions has been studied on rat uteri excised during pro and metaoestrus. At the concentration of 5 x 10-6M, the compound does not alter the spontaneous contractions of proestrus uteri and suppresses the phasic component of PGF-2alpha-elicited contractons. At the concentration of 5 x 10-5M, 7-oxa-13-prostynoïc acid inhibits spontaneous contractions, and inhibits PGF-2alpha-elicited contractions. In metaoestrus uteri, the inhibitory effect is not competitive. 7-oxa-13-prostynoïc acid is also able to inhibit non competitively the contractor effect of angiotensin II in proestrus uteri. This result, together with those of previous investigations performed using other prostaglandin inhibitors, suggest that the contractor effect of angiotensin II on the uterus is partially mediated by endogenous prostaglandins.


Pre and post ovulatory changes in the concentration of prostaglandins in rat graafian follicles.

The concentrations of prostaglandin F (PGF) and prostaglandin E (PGE) were measured by radioimmunoassay in isolated Graafian follicles of mature female rats during the pre and post ovulatory period of the estrous cycle. The levels of these prostaglandins were low and relatively constant from 8 a.m. to 4 p.m. on the day of proestrus, but there was a marked increase at 8 p.m. of proestrus reaching an apparent maximum at midnight (PGF 18 fold, PGE 70 fold). By 4 a.m. to 8 a.m. on the morning of estrus these prostaglandins declined rapidly to levels similar to those observed between 8 a.m. and 4 p.m. on the day of proestrus. The increases in prostaglandin levels occurred after the LH peak and apparently before the time of ovulation. These data confirm the role of PGF and PGE in the local mechanism of ovulation in the normal adult of a spontaneously ovulating animal species.


Influence of prostaglandin F autoantibodies on the estrous cycle of the guinea pig.

Adult female guinea pigs were actively immunized with prostaglandin F-2alpha conjugated to bovine serum albumin (BSA). Control animals, immunized against BSA continued to cycle normally, while the animals immunized against prostaglandin F-2alpha stopped cycling after one to three normal cycles. Laparotomy at 30 days after the last estrus revealed no recently formed corpora lutea. During the remaining 70 days of observation the antibody titer increased to 1:700, accompanied by increasing total serum estrogens (136 pg/ml at day 100) and a slow decline in circulating progesterone levels (0.6 ng/ml at day 100). The ovaries at day 100 contained degenerated corpora lutea and luteinized follicles. The suppression of the estrous cycle in the present experiments was interpreted as resulting from prolongation of luteal function as well as from inhibition of ovulation.


Prostaglandin specific binding in hamster myometrial low speed supernatant.

A charcoal adsorption method was developed to measure specific prostaglandin binding in low speed supernates of hamster myometrial homogenates. This method was used to characterize and quantitate PGE-1-specific binding. The equilibrium binding constants and the concentration of specific PGE-1 binding sites were determined during the hamster estrous cycle. The apparent association constant for 12 different preparations was 1.16 plus or minus 0.08 times 10-9M-1. The concentration of PGE-1 specific binding sites was significantly higher on days 2 and 3 of the estrous cycle that it was on days 1 or 4. The competition for PGE-1 binding sites by PGE-2, PGF-2alpha, tpga-1 and various PGE-1 metabolites and derivatives was measured in hamster myometrial homogenates. Relative affinities of the natural prostaglandins for the PGE-1 binding sites, calculated by parallel line assay, were: PGE-2 greater than PGE-1 greater than PGA-1 greater than PGF-2alpha. For PGE-1 metabolites the relative affinities were: PGE-1 greater than 13,14-dihydro-PGE-1 greater than 13,14-dihydro-15-keto-PGE-1 greater than 15-keto-PGE-1. For the analogs and derivatives the compounds tested ranked as: 16,16-dimethyl-PGE-1 greater than PGE-1 methyl ester greater than 17-phenyl-18,19,20-trinor-PGE-1 greater than 15(S) 15-methyl-PGE-1 methyl ester. Arachidonic acid, bis-homo-gamma-linolenic acid and 7-oxa-13 prostynoic acid had relative affinities greater than 0.1 compared to PGE-1 equal 100. Indomethacin had a relative affinity of 0.4 compared to PGE-1.


Prostaglandin-induced regression of porcine corpora lutea maintained by estrogen.

Corpora lutea were marked with suture in 24 crossbred gilts on day 7 to 9 of the estrous cycle (first day of estrus equals 0). All gilts were injected with 5 mg of estradiol benzoate (EB) daily from day 10 to 15 to extend the lifespan of corpora tutea, then the gilts were randomly assigned to two groups. On day 20, the 12 gilts of Group 1 were injected with 10 mg PGF-2ALPHA, and the 12 gilts of Group 2 were injected with saline. Ovaries were recovered 10 to 13 days after PGF-2ALPHA or saline injection. Ten gilts in Group 1 displayed estrus 5 plus or minus 0.7 days after PGF-2ALPHA injection, but only two gilts in Group 2 displayed estrus during the experimental period. In gilts that displayed estrus, all marked CL had regressed. Marked CL were still present in all 12 gilts that failed to exhibit estrus during the experimental period. These results show that in the pig, PGF-2ALPHA caused regression of CL that were maintained beyond the normal luteal phase of the estrous cycel by EB treatment.


Immunoglobulin clases IgG, IgM, IgA and complement component C3 in dental periapical lesions of patients with rheumatoid disease.

To identify the immunoglobulin classes present in dental periapical lesions and to determine how, if at all, the distribution of immunoglobulin classes IgG. IgM and IgA differ in the dental periapical lesions of rheumatoid and control patients, paraffin-embedded and frozen tissues from the dental periapical lesions of 36 rheumatoid and 22 control patients were examined by the direct immunofluorescence technique. In addition, both rheumatoid and control tissues were examined for the presence of complement component C3, albumin and fibrinogen. Neither the greater frequency and abundancy with which IgG and to a lesser degree, IgA occurred in the dental periapical lesions of rheumatoid patients than in the corresponding lesions of control patients, nor the more frequent occurrence of complement component C3 in these same lesions, was statistically significant. No difference in the distribution of the various immunoglobulin classed could be discerned between the rheumatoid and control group. Albumin was distributed almost equally between the two patient groups. The greater abundance of fibrinogen among control patients was, however, statistically significant. These results do not provide evidence that an immunologic reaction takes place in the dental periapical lesions of patients with rheumatoid disease similar to that which characterizes rheumatoid inflammation. However, the presence of such a reaction cannot be excluded without more extensive investigation.


Arteriovenous fistula with bovine arterial grafts for hemodialysis.

Bovine arterial graft arteriovenous (AV) fistulas have proven to be very helpful devices for access to the circulation in hemodialysis patients, especially after conventional AV shunts and fistulas cannot be made.


An evaluation of the radiological signs of fetal death.

Radiological investigation is warranted in the antenatal patient only if the findings are likely to influence future management. The major radiological signs of fetal death are gas in the fetus, overlapping of the cranial bones and Deuel's halo sign. Gas patterns in the portal and umbilical vessels are unique and pathognomonic. Deuel's halo sign would be found more commonly if the correct radiographic view was employed. Spalding's sign is of no value before 20 weeks or if the fetal head is subjected to pressure. Hyperflexion of the fetus more often indicates intra-uterine growth retardation than death in utero.


Determination of portacaval shunt patency in rats by splenoportography.

Open splenoportography was performed on a series of control, "sham-operated" and portacaval-shunt rats. Relevant vascular anatomy was well demonstrated. The criteria for diagnosis of patency, partial and total occlusion of portacaval shunts are mentioned and findings are correlated with postoperative weight variations. The possible significance of collaterals in the region of the shunt is briefly discussed.


Dextran and the prevention of postoperative thromboembolic complications.

1. The purpose of fluid administration is not only the restoration of blood volume but also the normalization of impaired nutritive flow. 2. Plasma oncotic (colloid osmotic) pressure is the only force which can draw water into the circulation. In shock the infusion of colloid solutions is able to normalize nutritive flow and peripheral resistance almost at once. 3. Five per cent solutions of pasteurized plasma protein or albumin and 6 per cent dextran 70 yield a volume expansion corresponding to the amount infused. 4. The decrease in hematocrit produced by the infusion of these three colloidal solutions is accompanied by a decrease in whole blood viscosity resulting in a rise in cardiac output as well as in nutritional tissue flow. 5. Hemodilution improves oxygen supply as long as the hematocrit does not fall below 30 per cent, although normovolemia is the critical requirement. 6. Transmission of viral hepatitis is still the greatest danger of blood transfusion. 7. The use of large amounts of Ringer's lactate is not advised, as this solution does not reduce the total number of units of blood which need to be given. Pulmonary edema may become a problem. 8. Dextrans are best suited to initial volume replacement in shock. They increase plasma volume, improve blood flow, have antithrombotic properties, and are easily available and relatively cheap. Anaphylactoid reactions are rare. 9. Every third patient undergoing general surgery and every other patient having hip surgery develops a deep venous thrombosis. Widespread prophylaxis to prevent thromboembolic complications is mandatory. 10. The antithrombotic properties of dextran are due to a reduction in platelet adhesiveness, a change in fibrin clot structure, and the increased lysability of thrombi and the improvement of blood flow. 11. In a personal controlled, prospective, randomezed trial comparing subcutaneous heparin and intravenous dextran 40, 35.8 per cent of the controls (n=95), 13.2 per cent of the 83 patients in the heparin group, and 20.5 per cent in the dextran group (n=83) developed deep venous thrombosis. The difference between dextran and heparin is not significant; however, both treatment groups show a statistically significant effect compared to the controls.


Occurrence of spontaneous hemorrhagic necrosis of the central nervous system in fetal hamsters.

Lesions of hamster fetal neuraxial tissues, characterized by multifocal and coalescent zones of hemorrhage, edema, and necrosis in the cerebral mantle, brainstem, and spinal cord, were observed in experiments designed to test the teratogenicity of potato preparations. Retrospective and prospective data indicated, however, that the potato preparations were not responsible but that the disease occurred spontaneously in the colony and was associated with direct breeding contact of virgin females with certain males. Observations suggest that an infectious agent may be responsible, but no agent was recovered. Immunofluorescence assay of inoculated cultures indicated that reovirus was not present in affected fetal tissues.


Studies on the platelet defect in alcoholism.

Platelet ultrastructure, protein composition, and adenine nucleotide metabolism were studied in patients ingesting ethanol to elucidate the mechanism of ethanol-induced changes in platelet function and survival. Serial measurements were made in 2 patients who maintained blood ethanol levels in excess of 300 mg/100 ml for 3 to 4 weeks. No major changes in structure or metabolism were detected in platelets from the patient whose platelet counts remained stable during the ingestion period. By contrast, the development of thrombocytopenia in the other patient was associated with significantly reduced intracellular ADP, increased ATP/ADP ratio, decreased release of ADP, increased specific radioactivity of intracellular ATP and ADP, and increased formation of hypoxanthine. Additionally, platelets from this patient varied markedly in size, contained giant granules, and possessed a poorly defined microtubular system. After stimulation with ADP or collagen, centripetal granule migration was retarded, and the aggregates formed were small and loose. Several large proteins were absent from the supernatant fraction of sonicated platelets from the thrombocytopenic patient. Exposure of normal platelets to ethanol in vitro resulted in no detectable change in platelet ultrastructure. The data indicate that the ethanol-related abnormalities of platelet function are due in part to subnormal amounts of intracellular ADP and a deficit in the storage pool of ADP. Additionally, the results suggest that impairment in the release mechanism to the observed defect in the release reaction.


Characterization of the fawn-hooded rat as a model for hemostatic studies.

A large colony of fawn-hooded (FH) rats, comprising five original families and six generations of their progeny, was developed for genetic and comparative studies of their bleeding tendency. The characteristics of the bleeding diathesis in these rats are similar to those originally reported in related rats by Tschopp and Zucker. FH rats have normal clot retraction, ADP-induced platelet aggregation and platelet ADP; variable aggregation with collagen; minimal aggregation with adrenaline and cobra venom factor; and reduced platelet ATP, ATP/ADP ratio, serotonin content and -14C-serotonin release. In comparison to age- and sex-matched Wistar rats, FH rats have significantly prolonged partial thromboplastin time, shortened Russell's viper venom time and increased factor X and XI levels. Other coagulation screening tests and specific assays for fibrinogen, plasminogen and factors VII, VIII and IX were normal. Some age- and sex-related differences in coagulation and other parameters were observed within each rat strain. Plasma proteins, glycoproteins and ceruloplasmin (copper oxidase activity) showed no abnormalities, nor did initial studies of immunoglobulins and complement. However, FH rats have significantly lower glucose and higher cholesterol levels than comparable Wistar rats.


Defaulters from follow-up after pre-partum cardiac failure.

The problem of default from follow-up prejudices the study of disease in Africa. During studies of 224 patients with pre-partum cardiac failure were found that 119 had defaulted from follow-up for six months or more, after an average period of 22 months from their admission. A sociologist tried to find all these patients at home and interviewed those who were found. If the patient was not found, information was sought from the village head and from friends and relatives. Thirteen patients had died. The 69 who were traced gave a total of 101 reasons for default. Domestic reasons prodominated, of which poverty, moving home and the difficulties of travel were the most important. Among personal reasons, feeling well was the commonest single reason, and was given by 19 patients. The results show that the problems of a subsistence agricultural economy dominate the reasons for default, and that such problem cannot be overcome without changes in society. Our findings are relevant in the care of all chronic diseases and throughout rural Africa.


Prevalence of diabetes mellitus among the ethnic groups in Singapore.

During a routine medical examination of 5280 government employees (2736 males, 2544 females; age range 17 to 66 years; 3386 Chinese, 1252 Malays, 508 Indians and 134 other ethnic groups) diabetes was found in 31 (0.59%). The prevalence was higher in males (0.95%) than in females (0.20%). The highest prevalence was in the age-group 50 to 59 years (5.4%); at ages 30 to 66 years, the overall prevalence was 2.18%. The prevalence of diabetes is higher in Indians (2.76%) than in Chinese (0.30%) and Malays (0.48%); the difference is statistically significant, as it also is in the age-group 30 to 66 years (Indians 6.36%, Malays 1.39%, Chinese 0.9%). Of the 31 cases, only four were previously known and only nine were overwieght. The possible reasons for the higher prevalence of diabetes in Indians are discussed.


Seasonal population densities of snails transmitting urinary and intestinal schistosomiasis in Lusaka, Zambia.

The two molluscan intermediate hosts of S. haematobium schistosomiasis: Bulinus africanus and Bulinus globosus, were found most commonly in habitats that retained water for a substantial part of the year rather than in temporary rain-filled pools. Biomphalaria pfeifferi which transmits S. mansoni were found principally in streams and impoundments like dams but scarce in rivers. In a permanent habitat, the peak density of snails was between the warm, dry months of August and September while in a temporary habitat B. (Physopsis) sp. were most abundant towards the end of the rainy season in March and April. The influence of rainfall and the prevailing the temperature on the fluctuations in the density of snails populations is considered. Further, the low infection rate in snails found in the field is discussed in relation to the critical effect of temperature on this feature. The problem of increasing migration of rural inhabitants to the peri-urban areas and the possible escalation of prevalence rates is considered in relation to the control measures that may be instituted in view of the substantial body of base-line data now available to effect focal control.


The proliferation of prostatic epithelium in chronic prostatitis.

The labelling index in chronic nonspecific prostatitis ranges from 0.7-2.1%. This value is markedly higher than in nodular hyperplasia (0.4%) as well as in differentiated adenocarcinomas (0.6%). Increased labelling index could only be determined in glands adjacent to inflammatory foci. In recognition of these findings we believe that a correct evaluation of the cell proliferation in chronic prostatitis is only possible if each single cell can be evaluated, whether it is altered by inflammation or not. Therefore, solid tissue biopsies appear to be more suitable in autroadiographical studies than cytological material alone.


Metastatic tumors of spermatic cord.

A case of metastatic carcinoma to the spermatic cord is presented. The previous 12 cases documented in the literature are reveiwed. In the majority of cases the primary tumor occurred in the gastrointestinal tract with metastasis to the spermatic cord by retrograde lymphatic flow.


The influence of dietary protein upon blood composition in dairy cows.

Diets of different protein content were fed to dairy cows in two experiments of seven months and 12 months duration.  Significant differences in the mean concentrations of serum urea, albumin and copper and of blood haemoglobin and packed cell volume were observed between cows receiving the various diets.  The greatest differences occurred when cows were in mid-lactation.  It was concluded that concentrations of urea in the serum of the cows were reflecting dietary protein intake and that concentrations of albumin, haemoglobin and PCV during lactation were affected by long-term protein status.  The possible significance of the results is discussed in relation to previous reports indicating interrelationships between protein and copper metabolism, and in relation to the recommended protein content of diets for cows.


On the specificity of mushroom Pleurotus ostreatus and Pleurotus spodoleucus extracts.

Extracts of mushrooms Pleurotus ostreatus and Pleurotus spodoleucus contain hemagglutinins with anti-H specificity similar to those found in Cytisus sessilifolius and Laburnum alpinum.


[The current place of diagnostic ultrasound in cardiology].

A non-invasive technique such as echocardiography has numerous advantages over invasive examinations of the cardiovascular system. Echocardiography involves no hazard or risk to the health of the patient, it can be repeated at any time and provides truly basal haemodynamic information by avoiding sedation, excitation of the patient and injection of dye. Problems arising from peculiarities in the behavior of ultrasonic energy and chest malformations in the patient have been largely overcome by technical improvements and a more sophisticated wasy of examination. The current diagnostic applications of echocardiography are reviewed on the basis of our own experience and the advantages and limitations of this method discussed.


[Modern methods in the diagnosis of breast disease (author's transl)].

240 patients attending a surgical clinic for breast diseases were examined clinically and subjected to mammography, thermography and ultrasonic investigation. Histological reports are available in 98 cases, enabling an evaluation of the accuracy of the used diagnostic methods. The most efficient diagnostic method is X-ray mammography, followed by thermography and ultrasound diagnosis, but these procedures are still fraught with methodological and technical problems.


Perforation of the uterus by the copper-t and copper-7 intrauterine contraceptive devices.

Uterine perforation in patients wearing the Copper-T and the Copper-7 intrauterine contraceptive devices has been studied. In Bollnäs, Sweden, three perforations occurred in 1 156 insertions of the Copper-7, and in New York, USA, six perforations occurred in 1 153 insertions of the Copper-T. Cervical perforation seems to be a special feature of the Copper-T, while the Copper-7 tends to perforate through the uterine wall. The perforations can be divided into "primary" perforations, related to the insertion procedure, and "secondary" perforations, caused by uterine contractions. The diagnosis and treatment of uterine perforations by intrauterine devices is discussed.


Idiopathic hypertrophic subaortic stenosis.

The clinical and laboratory findings in 29 patients with idiopathic hypertrophic subaortic stenosis are presented. Dyspnoea during exercise, angina pectoris, syncope combined with left ventricular hyperthrophy on ECG and chest X-ray and a systolic ejection murmur at the apex and the left sternal border are the most important findings. The findings were different in patients below and above 30 years of age. Most of the patients below 30 were in function group I, had a normal heart volume on chest X-+ray, and syncope was related to exercise. All patients above 30 had symptoms, nearly all were in function groups II-IV and often complained of palpitations, had increased heart volume on chest X-ray, sign of enlarged left atrium or atrial fibrillation of ECG. Syncope was not related to exercise, but always associated with palpitation in patients above 35 years of age. Pathologic Q waves were found more often in the younger age group. The differential diagnosis is discussed in relation to fixed aortic stenosis, mitral valve disease, ventricular septal defect, coronary artery disease, and hypertrophic cardiomyopathy without outflow tract obstruction.


Prednisone-induced hemostasis in a platelet function abnormality.

A 6-year old girl had chronic thrombocytopenia and substantial platelet function abnormality associated with detectable IgM antiplatelet antibody. Treatment with prednisone was followed by an improvement in bleeding time, although in vitro platelet function deteriorated. This observation suggests an effect of prednisone on hemostasis that is independent of platelet function. Further trials of prednisone in patients with similar platelet function abnormalities who have a substantial bleeding tendency would appear indicated.


Health care in America-what does the future hold?

Problems facing health care delivery in America and possible solutions are discussed. Specific issues covered are availability of health care services; health manpower; interaction of consumers and health providers in making decisions; health planning and regulation; preventive medicine and health education of the public; and comprehensive primary care. Of particular interest to pharmacists, the author discusses future division of responsibility among health professionals.


Intestinal anisakiasis. First case report from North America.

The first case of intestinal anisakiasis in North America is described. This parasitic disease is recognized as a public health hazard in Japan and Europe. Man becomes infected with a larval form of the nematode Anisakis by consuming raw or undercooked fish containing the parasite. Typically, patients present with acute abdominal syndromes. Clinical and reontgenographic features may cause confusion with regional enteritis. Histologically, a striking oesinophilic granulomatous reaction occurs. Anisakiasis is most effectively prevented by discouraging the consumption of raw fish.


Chronic superficial keratitis in dogs: detection of cellular hypersensitivity.

Dogs affected with chronic superficial keratitis (CSK) and clinically normal dogs were tested for cellular hypersensitivity, using the leukocyte migration-inhibition (LMI) technique to 3 ocular antigens (Staphylococcus aureus and corneal and iridal proteins). Affected dogs had statistically significant increases in hypersensitivity cellular responses against corneal and iridal antigens. Affected dogs did not differ from clinically normal dogs in their cell response to S aureus.


Efficacy of bunamidine hydrochloride against immature and mature stages of Echinococcus granulosus.

The efficacy of bunamidine hydrochloride (given at dose levels of 25 and 50 mg/kg of body wt) against immature and mature Echinococcus granulosus tapeworms was studied in experimentally infected dogs. The compound had average efficacies of 85.9 to 98.8 percent against the immature stages and was completely efficacious (100 percent clearance) against the mature worms. These data indicate that bunamidine HCl at the given doses can be used as a control measure against hydatid disease in the United States.


Temporal sequence of events in the estrous cycle of the bitch.

Serum concentrations of luteinizing hormone (LH), estradiol-17 beta, and progesteron were compared with the onset of diestrus in Beagle bitches. The LH peak occurred 8.0 (standard error (SE), 0.3) days before the onset of diestrus. Vaginal cytologic characteristics during proestrus were studied, and vaginal cornification was complete 12.1 (SE,0.2) days before the onset of diestrus. A time sequence of hormonal, vaginal cytologic, and developmental events was outlined during proestrus, estrus, and early diestrus.


Transient neonatal hypocalcemia, hypomagnesemia, and high serum parathyroid hormone with maternal hyperparathyroidism.

Findings in a case of neonatal tetany in a child whose mother had elevated blood parathyroid hormone levels because of a parathyroid adenoma are given. The child was found to have hypocalcemia, hypomagnesemia, and hyperphosphatemia. Parathyroid hormone in the infant was found to be elevated. It has been postulated that the pathophysiologic mechanism of the hypocalcemia in this syndrome is transient hypoparathyroidism. The present case suggests a different mechanism: hypomagnesemia and end-organ refractoriness to parathyroid hormone, a state analogous to pseudohypoparathyroidism.


Idiopathic hypertrophic subaortic stenosis in pregnancy.

The coexistence of pregnancy and idiopathic hypertrophic subaortic stenosis is a potentially dangerous combination. We report a 23-year-old white woman with idiopathic hypertrophic subaortic stenosis and pregnancy who presented with severe symptoms (Class IV) and modest outflow obstruction associated with marked mitral regurgitation. After delivery, the evidence for significant mitral regurgitation regressed, while the outflow obstruction seemed unchanged. However, she returned to Functional Class II. We review the mechanisms by which pregnancy and labor may alter the hemodynamics of idiopathic hypertrophic subaortic stenosis and we discuss recommendations for the management of these patients during pregnancy, labor, and the immediate postpartum period. We conclude that despite increasing symptoms, most women with diopathic hypertrophic subaortic stenosis can tolerate pregnancy and a vaginal delivery.


Phagocytosis of blood cells by splenic macrophages in thrombotic thrombocytopenic purpura.

A 43-year-old woman who developed thrombotic thrombocytopenic purpura was treated successfully with high doses of prednisone and a splenectomy. Imprints from the cut surface of the spleen showed marked phagocytosis of all the cellular elements of the blood. This observation suggests that hypersplenism, in addition to vascular occlusion, may play a major role in the physiopathology of some cases of thrombotic thrombocytopenic purpura.


Pinwheel rubeosis iridis following argon laser coreoplasty.

Rubeosis iridis developed in the right eye of a 10-year-old boy with Marfan's syndrome. This eye had a chronic retinal detachment which we could study adequately only after coreoplasty of the iris was carried out with the argon laser. The purpose of this paper is to describe our experience using the argon laser for coreoplasty, and to report the unique pinwheel configuration of the iris neovascularization that developed around the laser lesions. We believe the rubeosis iridis was most likely a sequelae of the chronic retinal detachment, and that the atrophic laser lesions probably acted as templates for the neovascularization, giving rise to its peculiar configuration.


Phenylthiocarbamide tasting in a sample of twins.

1. Twenty-eight pairs of monozygotic and eighteen pairs of dizygotic twins were tested for their ability to taste phenylthiocarbamide (P.T.C.) by the method of Harris & Kalmus (1949). 2. Much greater variance in threshold levels was detected within the DZ than within the MZ pairs and this confirms the genetic origin of most variation in ability to taste this substance. 3. Variance in threshold levels within MZ pairs is of the same magnitude as the variance of reported duplicate measurements on the same individuals. 4. DZ twin pairs were classified as either concordant or discordant in their tasting ability. Variance within concordant DZ pairs is significantly greater than within the MZ pairs and it is shown that this difference can be accounted for in terms of the incomplete dominance of the T allele reported by Kalmus (1958). However, the data do not exclude the possiblity that this greater variation in threshold levels is partly due to multiple alleles or background genetic variation. 5. The frequency of the t allele in Australians of European descent in estimate at 0-52 plus or minus 0-06.


Sibships ascertained through a twin pair with at least one affected twin.

For sibships ascertained through a twin-pair where at least one of the twins is affected, an exact conditional test is developed for the hypothesis that the probability for a dizygotic twin to be affected is equal to that of single-born children against the alternative that the probability is higher among twins. Such a situation may occur in disorders where a twin pregnancy may cause sporadic cases of non-genetic origin. The method has been applied to data on febrile convulsions. It is shown that dizygotic twins with a family history of the disorder are more likely to get febrile convulsions than their single-born sibs. I want to thank DR M. Lennox-Buchthal, M.D., Copenhagen, for providing me with data and histories.


[Results of the long laboratory storage of Streptoverticillium hachijoense (Jamaguchi) Baldacci, the producer of trichomycin].

Viability, morphologo-cultural features and antibiotic properties of Sv. hachijoense, strain LIA-0052 stored for 10 years in a dry state and in the state of a resting culture were studied. Spores and mycelium of 2-week strains most stable to some chemical and physical factors were used for drying. It was found that viability of strain LIA-0052 was maintained for a longer period of time after lyophilization, in garden soil and agar culture under a layer of mineral oil. By the end of the observation period the viability of the soil culture decreased and the morphologo-cultural properties were stabilized. When the strain was cultivated on media with sucrose, the level of its antibiotic activity increased.


Amikacin: a rapid and sensitive radioimmunoassay.

A rapid, sensitive, and specific radioimmunoassay has been developed for the measurement of amikacin, a new semisynthetic aminoglycoside antibiotic. Antisera were produced by immunizing rabbits with amikacin conjugated to porcine thyroglobulin. Sera were screened for amikacin antibodies with (3)H-labeled amikacin. Free and bound radioactivity were separated by the second antibody method. As little as 5 ng of the antibiotic can be measured in serum, cerebrospinal fluid, and urine. The standard curve is unaffected by pH (5-11), ionic strength (0.01 M to 0.3 M), MgCl(2) or CaCl(2) (1 to 8 mM/liter), or urea (10-300 mg/100 ml). The antibodies raised to amikacin are not able to distinguish between kanamycin and amikacin. The cross reactivity of all other antibiotics tested was <0.1%. Co-incubation of first and second antibody allows for a 30-min incubation period, making the assay useful for stat determinations. The standard curve is linear on a logit-log plot, allowing for rapid computer analysis of the standard curve, interpolation of sample values, and quality-control monitoring.


Identification of Harper-Cawston factor as thymidine phosphorylase and removal from media of substances interfering with susceptibility testing to sulfonamides and diaminopyrimidines.

Rich media support the growth of bacteria in the presence of concentrations of sulfonamides and diaminopyrimidines that are highly inhibitory when the organisms are grown on minimal media. Many such rich media can be made more suitable for susceptibility testing by the incorporation of lysed horse blood. Harper and Cawston characterized the active substance, Harper-Cawston factor (HCF), and later studies indicated it to be a protein. It has now been identified as thymidine phosphorylase. The identification follows from the identical purification pattern of HCF and thymidine phosphorylase activities from horse blood to a high degree of purity. Blood of goats, sheep, oxen, geese, chickens, cows, dogs, rats, and humans had neither biological activity. The identification of HCF as thymidine phosphorylase is consistent with the earlier findings of Koch and Burchall (1971) that most of the interfering effects of rich media could be accounted for by their thymidine contents, and that thymidine is much more active in this respect than is thymine.


Acute pelvic inflammatory disease and clinical response to parenteral doxycycline.

The bacteriology of acute pelvic inflammatory disease (PID) and clinical response to parenteral doxycycline were evaluated in 30 patients. Only 3 of 21 cul-de-sac cultures from PID patients were sterile, whereas all 8 normal control subjects yielded negative results (P< 0.005). Poor correlation was observed between cervical and cul-de-sac cultures. Neisseria gonorrhoeae, isolated from the cervix in 17 patients (57%), was recovered from the cul-de-sac only once. Streptococcus, Peptococcus, Peptostreptococcus, coliforms, and other organisms normally present in the vagina were the predominant isolates recovered from the cul-de-sac. Parenteral doxycycline resulted in rapid resolution of signs and symptoms (within 48 h) in 20 of 27 evaluable patients (74%). In five others, signs and symptoms of infection abated within 4 days. The remaining two patients failed to respond; in both cases, adnexal masses developed during doxycycline therapy. Gonococci were eradicated from the cervix in all but one patient who, nevertheless, had a rapid defervescence of symptoms. There was no clear-cut correlation between the clinical response and in vitro susceptibility of cul-de-sac isolates to doxycycline. These data confirm the usefulness of broad-spectrum antibiotics in acute PID. Culdocentesis is a reliable means of obtaining material for the bacteriological diagnosis of acute PID; however, the pathogenetic role and relative importance of gonococci and various other bacteria in acute PID need to be clarified further.


Bilateral carotid aneurysms secondary to radiation therapy.

A patient was seen with symptomatic, rapidly expanding aneurysms that developed in both carotid arteries 4 years after bilateral radiation to the neck, left combined mandibular resection, and radical neck dissection. The presenting symptoms were pain and transient ischemic attacks of cerebrovascular insufficiency. The aneurysms were treated uneventfully be resection and vein-graft replacement at 15-day intervals. Microscopy demonstrated typical radiation changes. Effects of radiation on arteries are reviewed.


A new urethroanal reflex revealed by sphincter electromyography.

Anal sphincter electromyography carried out during dilatation of the anesthetized urethra revealed a reflexogenic zone at the bladder neck, dilatation of which causes contraction of the anal sphincter. This reflex appears to be separate and distinct from previously described reflexes of the bulbocavernosus type which are triggered by somatic sensory stimuli. A possible role of this reflex in the separation of the functions of micturition and defecation is postulated.


Determination of the buoyant density of canine distemper virus by radioassay.

Canine distemper virus was labelled with tritiated uridine and, following precipitation with saturated ammonium sulphate solution, was concentrated 66-fold by centrifugation through a discontinuous sucrose gradient. When this preparation was centrifuged to equilibrium in density gradients of potassium tartrate or sucrose, radioactivity was distributed over the density range 1.218 to 1.180 with a pronounced peak at around 1.195. This corresponded closely to the distribution of infectivity and also to that of virus particles revealed by electron microscopy. In density gradients of caesium chloride, a plateau of radioactivity was present over the density range 1.26 to 1.24 with a peak at around 1.240 but most of the infectivity was limited to the range of 1.24 to 1.22. Since the amount of infectious virus recovered from potassium tartrate was greater than that recovered from the other two materials and the radioactive peak occurred over a narrower density range, it was concluded that potassium tartrate was the material of choice for the isopycnic centrifugation of canine distemper virus.


The growth of African horse-sickness virus in embryonated hen eggs and the transmission of virus by Culicoides variipennis Coquillett (Diptera, Ceratopogonidae).

Seven-day-old embryonated hen eggs were infected with African Horse Sickness virus by the yolk sac and intravenous routes. Virus reached a high titre in the blood of infected embryos. Culicoides variipennis midges which took a blood meal from infected eggs became infected with virus, and after 7 days at 26 degrees - 27 degrees C transmitted African Horse Sickness virus to uninfected eggs. C. variipennis may therefore be considered a biological vector of African Horse Sickness virus in the laboratory.


The multiplication of African horse-sickness virus in two species of Culicoides (Diptera, Ceratopogonidae).

Type 9 African horse-sickness virus multiplied to a high titre in both Culicoides nubeculosus and C. variipennis after intrathoracic inoculation and in C. variipennis after oral ingestion. The orally infected C. variipennis were able to transmit the virus by biting after 13 days incubation at 26 degrees C but not after 6 days incubation. Intrathoracically inoculated C. variipennis were able to transmit the virus after 4 days incubation. It is suggested that C. variipennis can act as a biological vector of African horse-sickness virus.


Brief hypoxia-ischemia initially damages cerebral neurons.

Rats were studied during cerebral hypoxic ischemia to determine whether neurons or blood vessels suffered the first damage. Ten or more minutes of unilateral carotid artery occlusion combined with systemic hypoxemia (PaO-2, 21 mm Hg) produced neuronal but not vascular damage in the ipsilateral cerebral hemispheres of 18 of 29 rats (62%); two and five minute stresses caused no visible neuronal abnormalities. The longer exposures produced more widespread damage, and neuronal loss and gliomesodermal reaction were evident after prolonged survival. Early neuronal changes correlated with abnormalities of motor behavior (P less than .005). Despite neuronal damage that was sometimes extensive, vascular no-reflow developed in only one of 24 animals after 20 and 30 minutes of hypoxia-ischemia. Production of neuronal and neurological abnormalities in the absence of hypotension or vascular no-reflow indicates that hypoxia-ischemia initially damaged cerebral neurons.


Conjunctivitis due to Thelazia californiensis.

A patient with chronic conjunctivitis was found to have a parasitic nematode infection of the conjunctiva. The parasite was found within the cul-de-sac of the conjunctiva, entirely motile. The treatment of the conjunctivitis was simple removal of the organism, and periodic follow up.


Choroido-vitreal neovascularization after argon laser photocoagulation.

Argon laser photocoagulation performed to destroy retinal neovascularization in two patients with proliferative sickle retinopathy and one patient with sarcoidosis was complicated by the development of choroidal neovascular tissue that extended through the photocoagulated retina into the vitreous. Attempts to obliterate the neovascular growth were successful in two cases. In the case that did not respond to repeated therapeutic procedures, the possibility of a direct communication with the long posterior ciliary artery was suggested by the presence of a chorioretinal defect in the vicinity of the photocoagulated area.


[Hereditary thrombocytopathic thrombocytopenia. Report of 2 cases transmitted with dominant character].

Hereditary thrombocytopathic thrombocytopenia is reported in a family (mother and son) and is transmitted with dominant character of moderate to severe intensity (platelets fluctuated from less than 5,000 per mm-3 to 20,000 with sporadic higher rises), normal survivorhship, larger than normal diameter and abnormal platelet 3 factor. Comments are made on its resemblance to idiopathic thrombocytopenic purpura and stress is placed on the importance of its identification through the decision of splenectomy with negative results in these patients.


[Hemangiolymphangioma of the abdominal cavity complicated with consumption coagulopathy].

A case of giant hemangiolymphangioma of abdominal cavity is reported. It showed severe bleeding from consumption coagulopathy and secondary fibrinolysis. (Kasabach-Merritt syndrome). A review is made of the production mechanism of coagulation abnormalities that may accompany vascular neoplasias, the usefulness of anticoagulants and the selective treatment in this type of disease.


Pathological features of hypertrophic obstructive cardiomyopathy (HOCM) in the elderly.

The pathological findings and available clinical data in 15 necropsy cases of HOCM, aged over 61 years, are reported. Three patients were in the eighth decade and 4 in the ninth; 8 were women. Five presented as sudden death, 2 died in congestive cardiac failure, and 7 died of unrelated conditions and HOCM was an apparently incidental postmortem finding. Compared with cases under 60 years, the hearts of the elderly patients were heavier and less likely to show typical asymmetrical hypertrophy, the free wall of the left ventricle also being thickened in two-thirds of the cases over 60 years. Most of the elderly cases showed a distinctive band of fibrous thickening over the upper part of the interventricular septum. This lesion had a "mirror image" relation to the lower part of the aortic surface of the anterior mitral cusp, with the histological features of a friction lesion. It appears to be a morphological expression of the systolic contact of anterior mitral cusp and interventricular septum seen on cineangiography and thus diagnostic of HOCM. Once formed, the fibrous band appears to persist even if the obstructive element disappears. It is, therefore, a valuable diagnostic feature indicating a diagnosis of HOCM in an age group where the morphology is usually not the classical asymmetrical form and in which this diagnosis is usually not considered.


[Congenital pyloric hyperthrophy. Factors associated with its frequency].

With the idea of clearing out several variables individually found associated with a greater incidence of congenital pyloric stenosis in the same group of children, 298 clinical histories in infants complaining of this deviation were reviewed. Information was obtained as to sex, birth number, blood group and the month these children with stenosis were born. The distribution of frequencies in each one of these variables was compared to that in 309 children surgically operated from peritonitis. The results showed a greater incidence of congenital pyloric stenosis in males than in females; also, the disease appeared more frequently in first-borns. Patients with type A blood group were less prone to present the anomaly. The birth month showed no statistical difference. The findings stress the importance shown by the genetic component of this disease.


Treatment of stress incontinence by a fibrin bioplast.

A new simple method of treating urinary stress incontinence is to fix a fibrin implant vaginally to the bladder neck and upper urethra. The support provided by the implant is maintained after its resorption by the fibrous tissue which replaces it. The two-year cure rate in 85 patients was 93 per cent.


Gynandroblastoma of the ovary.

A gynandroblastoma of the ovary in a 17-year-old girl is reported. Primary amenorrhoea, hirsuties and slight clitoral enlargement were associated with the tumour and high levels of circulating testosterone were found. This is the first gynandroblastoma in which Reinke crystalloids have been identified in the Leydig cell component. The criteria for the diagnosis of this tumour and its histogenesis are discussed.


The obstetric performance of nigerian primigravidae aged 16 and under.

The obstetric performance of 105 primigravidae aged 16 or under who received inadequate antenatal care was compared with that of 95 primigravidae of the same age who had adequate antenatal care and with 100 primigravidae aged 22 years who also had adequate antenatal care. The increased incidence of pre-eclampsia and babies weighing under 2-5 kg. in teenage pregnancy was confirmed. The effects of both hazards were diminished by good antenatal care.


5'-Nucleotidase from smooth muscle of small intestine and from brain. Inhibition of nucleotides.

5'-Nucleotidase prepared from muscle of small intesting of pig is strongly inhibited by nucleoside di- and triphosphates and their phosphonate analogs. Substrate kinetics appromate the Michaelis-Menten for for AMP, which shows a Km of 3-6 muM at pH 5.3-7.2. Inhibition is characterized as partial competitive, except at pH 5.3, where inhibition by ATP is noncompetitive. The Ki values for several inhibitors have been determined, and their departure from completeness of competitive inhibition has been studied. Inhibitor cooperativity of the type reported for the enzyme from sheep brain (P. L. Ipata (1968), Biochemistry 7, 507) was not observed for the enzyme from gut. In addition we failed to confirm sigmoid inhibition kinetics with 5'-nucleotidase from sheep brain.


Acetylcholinesterase-catalyzed hydrolysis of an amide.

In this paper we report that acetylcholinesterase catalyzes hydrolysis of amides, an observation which had not been made previously. The amide used is an analog of acetylcholine, 2-acetoaminoethyltrimethylammonium iodide. The experiments were performed with an enzyme preparation obtained from electroplax of Electrophorus electricus. Inhibition of the enzyme by a specific organic phosphate inhibitor abolished both the esterase and the amidase activity of the enzyme. The effect of hydrogen ions between pH 5 and pH 10 on the steady-state kinetic parameters, Km and kcat, has been investigated. These parameters show essentially the same dependence on pH as is observed in catalytic hydrolysis of acetylcholine. k-cat is controlled by an ionizing group of the enzyme with an apparent pK of approximately 6.3, and reaches a pH-independent maximum value of 3.6 sec- minus 1 above pH 8. The value for Km of 1 mM at pH 7 and 25 degrees is about five times greater than that for catalytic hydrolysis of the ester at the same pH and temperature. Preliminary electrophysiological experiments indicate that the amide analog binds to the receptor less well, by several orders of magnitude, than acetylcholine does.


Modification of pig heart lipoamide dehydrogenase by cupric ions.

The insertion of a second disulfide bridge into native pig heart lipoamide dehydrogenase, requires two Cu-2+ ions for each catalytic center inactivated under anaerobic conditions. During inactivation, both metal atoms become reducible by their juxtaposition to the two participating cysteine residues and may be removed as the Cu+-chelates of neocuproine and bathocuproinesulfonate, leaving an additional disulfide bridge on the protein. Inactivation does not require the presence of oxygen, but when substoichiometric levels of copper are used under aerobic conditions the slow regeneration of Cu-2+ becomes rate-limiting. The course of aerobic inactivation is markedly biphasic at 0 degrees using 2 Cu-2+/FAD, with 30% of the total change completed rapidly, followed by a much slower phase. Both the extent of the fast phase and the rate of the second phase are enhanced by increasing levels of Cu-2+, but are relatively unaffected when the Cu-2+/FAD ratio is maintained at 2 and the protein concentration is varied. The enzyme affords several binding sites for Cu-2+ at pH 7.8, and it is suggested that competition between these sites during the initial statistical distribution of metal ions may explain this biphasic behavior.


Lipid composition of the Golgi apparatus of rat kidney and liver in comparison with other subcellular organelles.

Golgi apparatus isolated from both rat liver and rat kidney have been characterized with respect to their neutral and phospholipid content and their phosphopipid composition and compared with mitochondria, rough endoplasmic reticulum and plasma membranes. In addition, the distribution of sulfatide in the subcellular fractions of rat kidney was determinich are rich in cholesterol esters and ubiquinone. Removal of about 75% of the cisternal contents of rat liver Golgi  reduced its content of cholesterol esters but not of ubiquinone. The Golgi complex of liver most closely resembles endoplasmic reticulum in its phospholipid composition except for a higher content of sphingomyelin. Removal of most of the contents of the Golgi cisternae did not appreciably alter the phospholipid composition of the Golgi apparatus of liver. Goligi apparatus from kidney has a phospholipid composition which resembles liver Golgi much more closely than it does any other cell fraction from kidney. The sulfatide content of kidney Golgi, the cell fraction richest in this glycolipid, is about 14% of the total lipid present in this fraction. Sulfatide was present in plasma membranes, mitochondria and rough microsomes, but at about one-third the level found in Golgi. Sulfatide is the main glycosphingolipid present in all the cell fractions from kidney which were studied.


Membrane-bounded lipid particles from beef heart acylglycerol synthesis.

The site for the synthesis of triacylglycerol in heart muscle has been the subject of this study. Acyl-CoA synthetase and glycerophosphate acyltransferase, enzymest necessary for the synthesis of triacylglycerol, have been found to be associated to membrane-bounded lipid particles. The acylglycerol synthesizing enzymes exhibit higher activities in the membrane-bounded lipid particles than in the mitochondrial or microsomal fractions. Futhermore, the lipid particles have the greater capacity for synthesis of diacylglycerols and triacylglycerols, whereas monoacylgly cerols are the major neutral lipid product in both the mitochondrial and the microsomal fractions. The results indicate that the membrane-bounded lipid particle may be the site for the synthesis as well as for the storage of intracellular triacylglycerol of the heart.


Physicochemical studies on tryptic digestion of bovine fibrinogen.

The high molecular weight fragments observed during tryptic digestion of bovine fibrinogen and the variation of their relative proportion with time has been studied. Separation of the different molecular species was carried out by gel filtration and the molecular weights of the isolated fragments were determined by sedimentation equilibrium and from their electrophoretic mobilities in sodium dodecylsulfate-polyacrylamide gels. The fibrinogen is degraded by trypsin into distinct fragments, with molecular weights of 270 000, 170 000, 90 000 and 50 000 accompanied by a series of smaller fragments whose properties were not investigated. The relative proportion of the components was estimated from area measurements on scans of the stained gels obtained after electrophoresis in the presence of sodium dodecylsulfate. The relative concentration and the molecular weight of each component established its molar concentration in each of the digestion mixtures obtained after varying incubation times (1-60 min). These data were used for a kinetic analysis of the process. The kinetic model derived on the basis of the trinodular model of fibrinogen (see Appendix) gave a very good representation of all the experimental results.


Changes in proteins of the human lens in development and aging.

A number of proteins have been isolated from the human lens at different stages of development, from before birth to old age. These proteins have been characterized and compared with each other and with corresponding proteins from bovine lens. Many similarities were found between human and bovine crystallins, but alpha-crystallin isolated from old human lenses using DEAE-cellulose, unlike bovine alpha-crystallin similarly isolated, is not found as large soluble aggregates. The amide contents of various lens protein fractions were determined. No extensive changes were found during adult life, but there was evidence that significant deamidation of alpha-crystallin had occurred before birth and possibly during infancy. The results are related to the unique development and aging of the lens.


Characterization of the collagen synthesized by cultured cartilage cells.

Cartilage cells from embryonic chick cartilage were grown in primary cultures. The cell layer was sequentially extracted with neutral saline, mercaptoethylamine and pepsin which revealed that these cells produced salt-soluble and salt-insoluble collagen. The alpha1- to alpha2-chain ratio was determined for the collagen extracted from the cultured cells and was found to be 13 to 1. Further analysis of the molecule was carried out by CNBr cleavage of the salt-extracted collagen and separation of resulting peptides by ion-exchange chromatography. It was shown that the cultured cartilage cells synthesize collagen of the type (alpha1[II])3.


Synthesis of immobilized flavin derivatives and their use in purification of chicken egg-white ovoflavoprotein.

Affinity adsorbents for flavoproteins were prepared by the covalent attachment of polyacrylamide and agarose to flavin derivatives linked through position N(3) of the flavin nucleus. 3-Carboxymethyl-FMN covalently linked to aminoalkyl substituted agarose was successfully used for the separation and purification of the apo form of the ovoflavoprotein from chicken egg white. High yields and high purities were achieved by two different isolation procedures employing the affinity adsorbent.


Nuclear magnetic resonance studies of hemoproteins. IV. Hindered rotation of heme side methyl group as a probe for studying van der Waals contacts in the heme side environments of myoglobin derivatives.

220 MHz roton NMR spectral evidence for restricted rotation of one methyl group in the heme side chain of ferric horse cyanomyoglobin is reported here. Temperature dependence of this methyl proton signal was computer-simulated, yielding 14,8 kcal/mol for the methyl hindered rotation. Ionic additives such as NaCl and (NH4) 2 minus SO4 caused a slackening of this restriction of methyl rotation, evidenced from collapse of methyl signal doubling by the addition of these ionic substances. This is discussed in terms of breaking of the salt bridge formed between one of the propionate COO minus group of heme and a part of the apoprotein which might lead to constraint of one of the heme side methyl groups. The peculiarity of hyperfine-shifted methyl proton signals for other myoglobin complexes such as azide and imidazole derivatives is also discussed briefly in terms of constraint of heme side methyl group buried in a hydrophobic cleft.


The primary sequence of chicken myoglobin (Gallus gallus).

After enzymatic digestion of chicken myoglobin by trypsin, chymotrypsin or thermolysin, the separation of peptides was performed by column chromatography on various ion exchange resins. Each peptide was purified by high-voltage paper electrophoresis or by chromatography either on paper or on ion-exchange resin, and its complete amino acid sequence was then determined by the combined dansyl-Edman procedure and by endopeptidase digestions. The whole globin was submitted to automatic Edman degradation using the Beckman sequencer. Residues have been positioned from overlaps of sequence data between tryptic (T), chymotryptic (C) and thermolysin (Th) peptides. The stepwise degradation of the whole globin confirmed the alignment of the N-terminal third of the molecule. The combination of these different approaches has led to the complete determination of the 153 residues sequence forming the polypeptide chain of chicken myoglobin. Comparison of the established chicken myoglobin structure with those from other species shows a conservation of structure, although the avian protein exhibits more variations in its amino acid sequence than has been found between other known myoglobins which all belong to mammalian species.


Involvement of oxyleghaemoglobin and cytochrome P-450 in an efficient oxidative phosphorylation pathway which supports nitrogen fixation in Rhizobium.

Cellular ATP level, ATP/ADP ratio and nitrogenase activity rise when oxyleghaemoglobin is added to respiring suspensions of Rhizobium japonicum bacteroids from soybean root nodules. Increased gaseous O2 tension is much less efficient than oxyleghaemoglobin in stimulation of bacteroid ATP production. Studies with the inhibitor carbonyl cyanide m-chlorophenylhydrazone show this ATP to be generated as a consequence of oxidative phosphorylation. N-Phenylimidazole, a specific cytochrome P-450 inhibitor, also lowers the efficiency of bacteroid oxidative phosphorylation. An approximately linear relationship is observed between ATP/ADP ratio and nitrogenase activity as N-phenylimidazole concentration is lowered. It is suggested that cytochrome P-450 is a component of the leghaemoglobin-facilitated respiration pathway and that it may act as intracellular O2 carrier rather than terminal oxidase. A less efficient oxidase appears to function when cytochrome P-450 is inhibited.


Differential labeling of glycerol moieties of phospholipids and triacylglycerols of cultured mammalian cells by [U-14 C] glucose.

Rabbit liver cells, in which fatty acid synthesis was suppressed by the rabbit serum component of the medium, were grown through 8- to 120-fold increases in cell numbers and mass of cell lipid in the presence of [U-14 C]-glucose. Triacylglycerols, phosphatidylcholine, and phosphatidylethanolamine were isolated from the total cell lipid and deacylated. Carbons 1 and 3 of the glycerol from the triacylglycerols and the no. 1 glycerol carbons of the two deacylated phospholids were oxidized by periodate and isolated as the dimedon derivative of formaldehyde. The specific activities of the glycerol carbons indicated that 58, 44, and 37 percent of the glycerol of the triacylglycerols. phosphatidylcholine, and phosphatidylethanolamine, respectively, were derived from the glucose of the medium. An additional 8 percent and 1-2 percent of the glycerol of each lipid was derived, respectively, from [U-14 C] glycerol and U14C-labeled amino acids added to the medium. In agreement with an experiment with albumin-bound [9,10- minus 3H]-oleic acid, and with smilar earlier experiments, it appears likely that appriacylglycerols originated from serum lipoproteins, or their partial hydrolysis products. An appreciable part of the ethanolamine of the cells' phosphatidylethanolamine originated from exogenous U- minus 14 C-labeled amino acids. Phosphatidyl-ethanolamine, however, was not a primary source of phosphatidylcholine. Labeling of the fatty acids of triacylglycerols and phospholipids by radioactive glucose, glycerol and amino acids was negligible.


Gangliosides of chemically and virally transformed rat embryo cells.

The gangliosides of control rat embryo cells, 3-methylcholanthrene, Rauscher leukemia virus, and combined 3-methylcholanthrene-Rauscher leukemia virus transformants were examined using [14 C]glucosamine as a tracer. All four cell lines exhibited a complex pattern of gangliosides. While N-acetylgalactosaminyl-(N-acetylneuraminyl)-galactosyl-glucosyl-ceramide was the major ganglioside in the control cell line, N-acetylneuraminyl-galactosyl-glucosyl-ceramide was the major ganglioside in the three transformants. The 3-methylcholanthrene transformant possessed a ganglioside pattern different from that of the Rauscher leukemia virus transformant. Hydrolysis of the gangliosides indicated that galactosamine, N-acetyl-and N-glycolylneuraminic acid were the labeled components in all cell lines.


Catabolism of an isolated, purified intermediate of prostaglandin biosynthesis by regions of the adult rat kidney.

1. A heat labile, cold-stable, stannous chloride-reducible intermediate of prostaglandin biosynthesis was formed in good yield (greater than 60%) from 3H-labeled arachidonic acid during brief incubations (30--90 s, 37 degrees C) with sheep seminal vesicle microsomes in the presence of p-hydroxymercuribenzoate (4 mM). This intermediate appears to have properties similar to one of the endoperoxides (15-hydroxyprostaglandin-9,11-endoperoxide) recently isolated by Hamberg and Samuelsson (Proc. Natl. Acad. Sci. U.S. (1973) 70, 889-903) AND Nugteren and Hazelhof (Biochem. Biophys. Acta. (1973) 326, 448-461). 2. Treatment of the purified intermediate with homogenates of rat kidney cortex, medulla and papilla resulted within 2 min (37 degrees C) in complete conversion into several compounds including prostaglandins E2 and F2alpha. The main product (40-50% yield formed by papilla homogenates was prostaglandin E2. The conversion into prostaglandin E2 was largely abolished by previous bo9ling of the homogenate whereas the conversion into prostaglandin F2alpha was not. The intermediate was stable in buffer for the same period of incubation. 3. The ratio of tritiated prostaglandins E2: F2alpha obtained were: papilla, 1.90; medulla, 0.76; cortex, 0.48. 4. These observations indicate that both types of prostaglandins can be formed by all three regions of the rat kidney and that regional differences exist in the proportion of E2 : F2alpha that is formed. Whereas prostaglandin E2 is mostly formed by an enzymatic process, prostaglandin F2alpha is not.


A pulse radiolysis study of some free radical reactions with erythrocyte membranes.

The reactions of the free radicals eaq- minus, OH and Br2- minus with haemoglobin-free erythrocyte ghost membranes have been studied by producing the radicals by pulse radiolysis and monitoring their reactions by optical spectroscopy. Hydrated electrons react rapidly with the membrane, but no attack at disulphide links was observed. Hydroxyl radical attack produced transient species absorbing weakly in the ultraviolet, which may arise from carbohydrate residues, such as N-acetyl neuraminic acid and N-acetyl glucosamine, on the membrane surface. No evidence was obtained for OH attack at ring-containing amino acid residues of the protein component. The Br2- minus radical, a more selective electrophile than OH, reacted only slowly with erythrocyte ghosts. Solubilization of the membranes with dodecylsulphate or digestion with alkali exposed protein containing tyrosine and tryptophan residues which reacted with Br2- minus. These results support other evidence for the absence of reactive protein at the membrane surface.


The independence of membrane potential and potassium activation of the sodium pump in muscle.

The membrane potential (Em) of sartorius muscle fibers was made insensitive to [K+] by equilibration in a 95 mM K+, 120 mM Na+ Ringer solution. Under these conditions a potassium-activated, ouabain-sensitive sodium efflux was observed which had characteristics similar to those seen in muscles with Em sensitive to [K+]. In addition, in the presence of 10 mM K+, these muscles were able to produce a net sodium extrusion against an electrochemical gradient which was also inhibited by 10- minus 4 M oubain. This suggests that the membrane potential does not play a major role in the potassium activation of the sodium pump in muscles.


Immunological evidence for the localization of sialoglycosphingolipids at the cell surface of sea urchin spermatozoa.

The localization of sialoglycosphingolipids in the plasma membrane of sea urchin spermatozoa was studied by employing immunological methods including immunolysis of liposomal model membranes. The antibodies produced against these complex lipids were found to agglutinate various sea urchin spermatozoa differently. Both species differences and species similarities in the agglutination were found in spermatozoa of the echinoderm, the sea urchin and the starfish. The agglutination of the sea urchin spermatozoa was inhibited specifically by ceratain carbohydrates. Only a limited number of molecular species of sialoglycosphingolipid were localized at the surface of the plasma membrane of sea urchin spermatozoa cells. Moreover, topographical differences were found in the localization of the sialoglycosphingolipids at the cell surface of spermatozoa.


Studies on the turnover of plasma membranes in cultured mammalina cells. I. Rates of synthesis and degradation of plasma membrane proteins and carbohydrates.

The rate of turnover of membrane proteins and membrane-bound carbohydrates in exponentially growing and in confluent contact-inhibited cultures of strain MK-2 cells was investigated. Cells were labelled with [14-C]leucine and [3-H]glucosamine, incubated in isotope-free medium and, at various times thereafter, the cells were harvested and membranes isolated from them. The rate of decay of the protein and carbohydrate components was determined from specific activity dilution of the labeled components in the isolated membranes. Although the rate of membrane synthesis is different in exponential and contact-inhibited cells, the rate of degradation (turnover) of membrane proteins and carbohydrates was found to be the same (25% per generation (42 h) or 0.6%/h).


Studies on the turnover of plasma membranes in cultured mammalian cells. II. Demonstration of heterogeneous rates of turnover for plasma membrane proteins and glycoproteins.

The relative rate of turnover of individual membrane proteins and glycoproteins in exponentially growing and contact-inhibited MK2 cells was investigated. Plasma membranes were isolated from cells that had been sequentially labelled with 14-C and 3-H isotopes of leucine and glucosamine. The membranes were then solubilized in sodium dodecylsulfate and their polypeptides separated by acrylamide gel electrophoresis. The 3-H/14-C ratios of the individual polypeptides reflected their relative rates of turnover. The proteins and glycoproteins of the exponentially growing cells exhibited markedly heterogeneous rates of turnover. In contrast, polypeptides in membranes of contact-inhibited cells exhibited a lesser degree of heterogeneity of turnover. In both exponential and contacted cell membranes a glycoprotein with a high apparent molecular weight exhibited the fastest rate of turnover.