[Effect of lecithin on liver microsomal lipid peroxidation]. The effect of exogeneous (egg) lecithin on peroxidation of microsomal lipids was studied with the view of elucidating the role of various components of lipid substrate in the overall oxidation rate of the lipids. The following processes were studied a) NADPH-dependent microsomal lipid peroxidation in the presence of lecithin; b) ascorbate-dependent microsomal lipid peroxidation in the presence of lecithin; c) oxidation of lipid mixture, isolated from the microsomes, and that of lecithin in the presence of the Fe2+ + ascorbate system; 4) oxidation of lecithin induced by the Fe2+ + ascorbate system. It was found that in the presence of exogeneous lecithin the oxidation of microsomal lipids in inhibited, which is probably due to the peculiarities of lecithin oxidation. It was shown that the specific rate of lecithin oxidation is decreased with an increase in lecithin concentration. Possible mechanisms of lecithin effect on microsomal lipid peroxidation are discussed.


[Principle of free energies linearity in chymotrypsin catalysis]. The applicability of the previously described principle of free energies linearity for semispecific substrates of chymotrypsin for the derivatives of acetylphenylalanine, acetyltryptophane and acetyltyrosine was demonstrated. The pH dependence of the parameters of a linear equation describing the relationship between the free energy of hydrolysis and the logarithmus of the kcat/Km(app) ratio suggests that at optimal pH the stability of the activation barrier of enzyme acetylation for all semispecific substrates is observed.


[Effect of some fragments of peptide hormones on the content of biogenic monoamines from mouse brain]. The effect of some tripeptides, which are fragments of peptide hormones, and their analogs on the content of biogenic monoamines (BM) from albino mice brain was studied. It was found that thyroliberin, melanostatin and the C-terminal tripeptide of gonadoliberin activate the dophaminergic (DA-ergic) system in the forebrain of mice treated with reserpine or haloperidol, whereas the C-terminal tripeptide of gastrin acts as a synergic blocker of the DA receptors. The N-terminal tripeptides (with and without the amido group) do not affect the content of BM. No effect of the tripeptides was observed in intact animals. It is assumed that the agonistic or antagonistic effect of the tripeptides on BM is due to certain structural peculiarities of the tripeptides, e.g. the presence of the C-terminal amido group and their endogenous nature.


Changes in the composition and properties of humic acids brought about by the action of microorganisms of the genus Nocardia. The results of an investigation of the influence of microorgansims of the genus Nocardia on changes in the composition and properties of humic acids are presented. According to these data, the elemental composition and IR spectra showed different effects on the structure of humic acids after utilization of these compounds as a nutrient source for three and six months by microorganisms.


Monoacylcadaverines in schizophrenia.

Monoacylcadaverines in schizophrenia. Monoacetylcadaverine and monopropionylcadaverine were found in the blood of schizophrenic patients in higher concentrations than in the blood of nonschizophrenic subjects. The blood levels of both monoacylcadaverines in schizophrenics were relatively higher during exacerbation of the illness and were relatively lower during improvement.


An abdominal extraperitoneal approach for the difficult orchidopexy. An abdominal extraperitoneal approach is described for use in patients who have a severe degree of incomplete descent of the testicle. In 86 such operations, the testicle was placed in the scrotum in 85. The approach allows certainty of diagnosis in cases of anorchia and it is helpful when orchidopexy has to be combined with a herniotomy in infancy.


Sequential patterns of haemodynamic and metabolic changes in experimental hypovolaemic shock. I. Responses to acute haemorrhage. Little is known about cardiorespiratory changes during the development of hypovolaemia. This study attempts to provide such information and compares the period of bleeding with that of established hypovolaemia. Eleven anaesthetized and ventilated greyhounds were bled and analyses of cardiopulmonary function made at fixed intervals both during and after haemorrhage. Six sequential patterns of cardiopulmonary and metabolic change were recognized. It was apparent that bleeding caused the first three phases of change, recovery from the effects of bleeding the next two and steady hypovolaemia the last. The event of bleeding is the main factor that elevates total peripheral resistance and reduces tissue perfusion with consequent lowering of oxygen consumption and alkalosis secondary to impaired carbon dioxide production; when bleeding ceases these changes partially reverse in a manner characteristic of that induced by the reinfusion of shed blood; and hypovolaemia per se has a relatively weak influence. These findings provide an explanation for disparities in previous published reports and have obvious clinical implications.


Comparative study of automatic blood-gas analysers and their use in analysing arterial and capillary samples. Three automatic blood-gas analysers were compared for ease of use; calibration; reproducibility and accuracy of results; maintenance; fault-finding; and use of expert technician time. Results obtained from arterial and capillary blood were compared with duplicate values obtained with a semi-automatic analyser controlled and calibrated with tonometered blood. No analyser was fully automatic, and all three needed maintenance by expert technicians. Difficulties were encountered when inexperienced operators used the machines. One automatic blood-gas analyser gave aberrant values for oxygen pressure (PO2) due to electrode dysfunction that was not indicated by the fault-finding system. A second analyser gave significantly lower values for blood pH than the standard machine. A comparison of pH, carbon dioxide pressure (PCO2), and PO2 measured in 40 simultaneous paired samples of arterial and arterialised capillary blood showed no significant difference for pH or PCO2, but the PO2 values were significantly lower in the capillary samples over the range studied. We conclude that all machines perform satisfactorily in terms of blood-gas analysis, but none may be regarded as fully automatic. Some degree of technical supervision is essential, as is proper training for all potential users.


Electron microscopic localization of substance P and enkephalin in axon terminals related to dendrites of catecholaminergic neurons. Morphological and pharmacological data suggest that catecholaminergic neurons receive afferent axons positively labeled for the peptides, substance P and [Met5]-enkephalin. In the present study, electron microscopic immunocytochemistry was used to determine whether a positive reaction for these peptides could be localized to axon terminals forming synapses with catecholaminergic neurons in the locus coeruleus and A2 regions of rat brain. Adjacent sections through these areas were incubated with antiserum to either substance P, [Met5]-enkephalin, or tyrosine hydroxylase, a specific marker for catecholaminergic neurons. The sections were subsequently processes by the peroxidase-antiperoxidase immunocytochemical technique. In both the locus coeruleus and A2 region, tyrosine hydroxylase was localized primarily to perikarya and dendrites of intrinsic neurons; whereas substance P and enkephalin-like immunoreactivity was localized to axons and axon terminals. The axon terminals showing positive reactions for substance P and [Met5]-enkephalin were morphologically similar to each other and to one type of axon terminal which formed synapses with dendrites labeled for tyrosine hydroxylase. This type of axon terminal always formed asymmetric synaptic junctions and contained 3-4 large (75-100 nm) dense vesicles (LDVs) and many small (40-60 nm) clear vesicles (SCVs). The reaction product for substance P and [Met5]-enkephalin was distributed throughout the lumen of the LDVs and formed a rim of labeling around the outer boundaries of the SCVs. These findings demonstrate that substance P and [Met5]-enkephalin-positive reactions are selectively localized to subcellular organelles in axon terminals in the locus coeruleus and A2 region of rat brain. They further suggest that the labeled axon terminals form synapses with dendrites of the catecholaminergic neurons.


Phenytoin and transmitter release at the neuromuscular junction of the frog. The effects of phenytoin (diphenylhydantoin, DPH) on transmitter release were studied at the frog neuromuscular junction. It was found that in Ringer's solutions containing a normal concentration of Ca2+ ions, DPH (1-2 X 10(-4) M) depresses neurally evoked transmitter release, whereas in Ca2+-deficient Ringer's solutions it produces an increase in evoked release. Spontaneous transmitter liberation is augmented by DPH under all the above conditions. An abrupt disappearance of the evoked response occasionally occured with stimulation at 0.5 Hz, but a normal response could be elicited by a second stimulus delivered shortly after the first. At 100-200 Hz, DPH regularly induced a partial block in synaptic transmission. In 8 mM MgCl2, this phenomenon appeared at 50 Hz and developed into a total neuromuscular blockade.


Correlation between presynaptic dense bodies and transmitter output at lobster neuromuscular terminals by serial section electron microscopy. Lobster neuromuscular terminals releasing comparatively small (low-output type) and large (high-output type) amounts of transmitter but arising from the single excitatory motor axon to the proximal accessory flexor muscle were serially sectioned for electron microscopy. The three-dimensional reconstruction showed the two types of terminals to have a complex branching pattern in which thin branches of the motor axon often enlarged into synapse bearing terminal regions. Quantitative comparison showed that the mean surface area of a synapse is similar in the two types of terminals. However, the low-output terminal has a higher synaptic density and devotes a greater part of its surface area to synapses compared to its high-output counterpart suggesting that transmitter output is not directly related to synaptic area. The mean surface area of a presynaptic dense body is not significantly different between low- and high-output synapses, but there is a significantly greater density of these active zones in the high-output terminal. This results in the ratio of mean dense body area to mean synaptic area being approximately 3 X greater in the high-output synapses than the low-output ones. This significant difference in the surface area of presynaptic dense bodies between low- and high-output synapses correlates with the difference in transmitter output at these two synapses, and implicates the dense bodies in the mechanism of transmitter release at lobster neuromuscular synapses.


Unit activity of limbic system neurons: effects of morphine, diazepam and neuroleptic agents. The effects of morphine, diazepam and three neuroleptic agents (chloropromazine, perphenazine and haloperiodol) on neuronal firing rats were studied in the limbic system of immobilized cats. Parietal craniotomy was carried out under 1.5--4.0% halothane. Extracellular potentials from single cells in the cingulate gyrus, septum and lateral hypothalamic areas were recorded using glass-coated, platinum-iridium microelectrodes. In general, intravenous adminstration of morphine sulfate augmented the spontaneous firing rates of most of the neurons studied. In contrast, diazepam produced a marked attenuation of both spontaneous and morphine augmented firing rates, whereas the neuroleptic agents had no significant or consistent effects on the morphine augmented firing rates of neurons in these limbic areas. These data indicate that the limbic system may play an important role in the behavioral excitement in cats induced by morphine administration and also the depressant effect of the tranquilizer diazepam. In contrast, the inability of the neuroleptic agents to antagonize the morphine augmented neuronal firing rates suggest these agents may act outside the limbic areas studied here.


Toleration of artificial tears - the effect of pH. We studied the effect of altering the pH of an artificial tear preparation on the tolerance of 20 patients with keratoconjunctivitis sicca. Of those who expressed a preference, 11/17 preferred a tear substitute more alkaline than any preparation now available in North America. We recommend an alkaline tear substitute as a valuable alternative in patients with keratoconjunctivitis sicca.


A study of carbachol-atropine interaction on intestinal smooth muscle vesicles, using a fluorescent probe. Plasma membrane vesicles were prepared from guinea pig ileum longitudinal muscle. The vesicles were characterized by electron microscopy and analysis of lipid and protein content. They were shown to be free of gross contamination from actomyosin, sarcoplasmic reticulum, and mitochondria. 8-Anilino-1-naphthalene sulphonic acid (ANS) binding characteristics were similar to those found in other membranes. Both carbachol and atropine increased the fluorescence of ANS bound to this membrane, the maximum increase for atropine being greater than that for carbachol. Since neither drug effected the apparent affinity constant for the ANS-membrane interaction. It may be assumed that the increased fluorescence was due to an increase in the number of ANS binding sites. The carbachol-dependent increase in ANS fluorescence was blocked noncompetitively by atropine but not by tubocurarine or diphenhydramine. These latter two antagonists also increased ANS fluorescence but at much higher concentrations than either carbachol or atropine. Neither atropine nor carbachol increased ANS fluorescence on either erythrocyte ghosts or liposomes (prepared from a lipid extract of the muscle membrane).


The effect of pH and amino acids on conidiation and pigment production of Monascus major ATCC 16362 and Monascus rubiginosus ATCC 16367 in submerged shaken culture. Monascus major ATCC 16362 and Monascus rubiginosus ATCC 16367 were cultivated aerobically on media containing nitrate or ammonium as nitrogen source to which the following modifications were made: (1) pH adjusted to 2.5 before sterilization; (2) addition of yeast extract; (3) addition of amino  acids in identical proportions and concentrations to those found in yeast extract; (4) adjustment of pH to 2.5 after addition of amino acids. The addition of amino acids in the form of yeast extract increased mycelium formation and reduced conidiation and pigment production. The addition of an amino acid mixture did not increase mycelium formation to the same extent as yeast extract but increased the number of conidia, while pigment production was reduced, especially when nitrate was the nitrogen source. As the amino acids are taken up after conidial formation has started, it would appear that it is not the amino acids themselves which are directly responsible for the induction of conidiation. The addition of amino acids inhibits nitrate and ammonium uptake suggesting the need for an early intracellular nitrogen limitation to induce conidiation. Lowering the pH inhibits the formation of conidia and increases pigment production; also the effect of amino acid addition is totally annulled. The pH of the medium is all important in regulating the formation of conidia and pigment production. The possible effects of the pH on the uptake of certain medium components is discussed, as well as their possible control of certain metabolic pathways which ultimately determines the availability of intermediates for conidiation and pigment production.


[Evaluation of the efficiency of the adsorption-elution technic for poliovirus 1 on fiber glass filters: application in the virological analysis of 100 ml to 1000 1 of water]. The efficiency of the adsorption-elution technique using fiber glass filters to concentrate viruses from water was evaluated to detect poliovirus type 1 in drinking, river, and sewage water. At pH 3.5 and with 5 X 10(-4) M aluminium chloride more than 99% were adsorbed at a 0.25-micron filter. Beef extract (3%), pH 9, eluted 85-95% of the adsorbed viruses and organic flocculation at pH 3.5 permitted to reconcentrate the viruses in 1/20 of the elution volume with a 50-72% efficiency. The overall efficiency of the technique for 100 ml to 1000 l of the different types of water using 10(2) to 10(6) PFU was 38 to 58%.


Physicochemical properties of Nebraska calf diarrhea virus hemagglutinin. Highly purified Nebraska calf diarrhea virus (NCDV) was prepared by cesium chloride density gradient centrifugation. The effect of temperature, pH, different concentrations of formaldehyde, chloroform, ether, ethyl alcohol, and methyl alcohol on NCDV hemagglutinin and virus morphology was studied. NCDV hemagglutinin was inactivated by temperature, pH 2.0, chloroform, ethyl alcohol, and methyl alcohol.


Functional implications of changes in the senescent brain: a review. The morphological, chemical, and physiological changes in the brain accompanying old age are reviewed. The deterioration of the striatal and hypothalamic dopaminergic systems were implicated in the onset of age related Parkinsonian-like slowing of performance and altered affect. Cholinergic hippocampal and neocortical systems were chemically and physiologically abnormal in the aged. The implications for slowed cognitive processing and persistance of the memory trace are presented.


Pneumococci resistant to erythromycin.

Pneumococci resistant to erythromycin. Susceptibility to erythromycin was determined for all pneumococci isolated in one laboratory from clinical specimens between 1969 and 1977. All 4724 isolates examined prior to October 1973 were susceptible to erythromycin. From October 1973 to December 1977, 64 (0.71%) of 8995 pneumococcus isolates were resistant to erythromycin. The resistant strains were isolated from 38 patients living in six widely separated communities in Alberta. The erythromycin-resistant strains were of nine capsular types, including six that often cause bacteremic disease and five for which resistance to erythromycin has not been reported hitherto. Certain strains of type 33 and of type 15 were highly resistant, the minimum inhibitory concentration (MIC) of erythromycin being 2000 microgram/mL; these strains were also highly resistant to lincomycin and clindamycin. Resistance in strains of other types was much lower, the MIC of erythromycin being 0.6 to 20 microgram/mL, and all but one of these strains were susceptible to lincomycin and clindamycin. All the erythromycin-resistant pneumococci were suspectible to penicillin.


Enzyme changes after coronary angiography. The enzymatic response to contrast media after coronary angiography was observed in 24 patients. The enzymes studied were glutamic oxaloacetic transaminase (SGOT), glutamic pyruvic transaminase (SGPT), lactic dehydrogenase (LDH), alpha-hydroxybutyrate dehydrogenase (alpha-HBDH), gamma-glutamyl transferase (gamma-GT), creatine phosphokinase (CPK), and the myocardial-specific isoenzyme of CPK (CK-MB). Angiography produced no significant elevations in SGOT, SGPT, alpha-HBDH, LDH, or CK-MB activities; gamma-GT increased up to the twelfth hour. High levels of CPK and CK-MB observed in one patient were found to be products of a thymus carcinoma. In vitro examinations also showed that concentrations of contrast media used in coronary angiography do not inhibit gamma-GT, LDH, and CPK activities.


Organization of haemopoietic stem cells: the generation-age hypothesis. This paper proposes that the previous division history of each stem cell is one determinant of the functional organization of the haemopoietic stem cell population. Stem cells from a lineage of stem cells which have generated many stem cells (older stem cells) are used in the animal to form blood before stem cells which have generated few stem cells (younger stem cells). The stem cell generating capacity of a lineage of stem cells is finite. After a given number of generations a stem cell is lost to the stem cell compartment by forming two committed precursors of the cell lines. Its part in blood formation is taken by the next oldest stem cell. We have called this proposal the generation-age hypothesis. Experimental evidence in support of the proposal is presented. We stripped away older stem cells from normal bone marrow and 13 day foetal liver with phase-specific drugs and revealed a younger population of stem cells whose capacity for stem cell generation was three- to four-fold greater than that of the average normal, untreated population. We aged normal stem cells by continuous irradiation and serial retransplantation and found that their stem cell generative capacity had declined eight-fold. We measured the stem cell generative capacity of stem cells in the bloodstream. It was a half to a quarter that of normal bone marrow stem cells and we found a subpopulation of circulating stem cells whose capacity for stem cell generation was an eighth to a fortieth that of normal femoral stem cells. This subpopulation was identified by its failure to express the brain-associated antigen which was present on 75% of normal femoral stem cells but was not found on their progeny, the committed precursors of granulocytes.


Neuroglandular contacts between vasopressin-reactive fibers and cells of the pars tuberalis in the rat. In electron micrographs fibers containing vasopressin-immunoreactive elementary granules 100--120 nm in diameter are observed within the basal lamina of the adenohypophyseal pars tuberalis adjacent to the rostral portion of the median eminence. The concept of a neuroglandular transmitter function of vasopressin is discussed.


Rapid method for measuring intracellular pH in vivo. Intracellular pH (pHi) was simultaneously measured in 6 normal tissues and a malignant tumour of rats by a rapid triple isotope technique, based on the in vivo distribution of 5,5-dimethyl-2,4-oxazolidinedione-2-14C (DMO), tritiated water and sodium chloride-36. Results compared favourably with pH measured directly in the same rat by capillary glass electrode, and with values of other workers for pHi in rat tissues. Mean pHi of normal tissues was close to pH 7, and in each organ there was a linear relationship between pHi and extracellular pH (pHe) over the normal range of pHe encountered (pH 6.9-7.6). Organ pHi altered in response to administration of NH4Cl or NaHCO3 to the host.


PH-dependent behavior of erythrocyte membrane elevations. II Occurrence in moderate acidemic and alkalemic conditions. Changes of the number of red cell membrane elevations revealed by freeze-etch electron microscopy are pH-dependent in vitro and in vivo. In healthy volunteers a decrease of the number of red cell surfaces with elevations was observed under moderate acidemic and alkalemic conditions. These changes were different in patients with Huntington's chorea and affective psychotic disorders. Elevations of the red cell membrane with the same diameter can be demonstrated in ultrathin sections, if sodium azide is added during the fixation procedure.


[A serological survey of arboviruses in the human population of Senegal]. 2,457 serum samples collected during an epidemiological survey on treponematosis in Senegal by the W. H. O. IR051 team in five areas of Senegal between 1972 and 1975 were studied for arbovirus antibodies using hemagglutination inhibition test. In addition 998 were studied using CF test and 1012 in neutralization test with yellow fever virus. The prevalence of arbovirus antibodies is important in all the country and does not vary basically between the different areas. In group A, chikungunya antibody is the most frequent and Sindbis antibody was found only in areas where migrant birds are resting. There is evidence of recent yellow fever infection in Upper Casamance and in Eastern Senegal. Most of the children are not immune and have to be immunized. Zika and Wesselsbron viruses show activity in all the areas and West-Nile, as Sindbis, is related to areas where birds are numerous. Bunyamwera virus activity is low.


Contact dermatitis from ethylenediamine. This study of seven patients with an allergic contact dermititis from ethylenediamine has shown that sensitisation to topical antihistamines and disodium ethylenediamine tetra-acetate is not inevitable in a allergic contact dermatitis from ethylenediamine.


The effects of bile on the gastric mucosa. Pure rat bile, mixed with an equal volume of 43% ethanol or distilled water, was intubated into the stomachs of fasted rats. Control rats were intubated with saline solutions or received no intraluminal solutions into the stomachs. Following a 10 min period of exposure of the bile or saline solutions to the luminal surface of the gastric mucosa, samples of the stomachs were processed for routine scanning and transmission electron microscopy and light microscopy. Observations indicated that the bile solutions produced extensive damage to the luminal surface of the stomach. The most prominent alteration was evident as a massive cytolysis of the epithelial cells which line the gastric glands, pits, and luminal surface of the gastric mucosa. Loss of the epithelium revealed the underlying honey-combed structure of the lamina propria. In addition, haemorrhagic areas were often observed throughout the mucosa. The findings of this study indicated that bile mixed with water or ethanol was highly destructive to the gastric epithelium, even more destructive than a solution of ethanol and water; however, the basal lamina and connective tissue fibres of the lamina propria were apparently unaltered by the bile solutions. Furthermore, the presence of an intact lamina propria in many areas where there was not extensive haemorrhage, following exposure of these solutions to the gastric mucosa, suggested the existence of a scaffolding structure for the proper orientation of the reconstruction process of epithelium of the glands, pits, and luminal surface of the altered gastric mucosa.


Laboratory studies with BL-S 578 (Cefadroxil) a new broad-spectrum orally active cephalosporin. BL-S 578 (Cefadroxil) is a new orally active semisynthetic cephalosporin antibiotic with broad-spectrum antibacterial activity. The new compound was evaluated in vitro in comparison with cephalexin. Some properties studies such as, antibacterial activity, binding with serum proteins and stability in acid and neutral solution at 37 degrees C for both cephalosporins were similar. In experimental infections of mice, the protective action of BL-S 578 was more effective than cephalexin against Staphylococcus aureus and Streptococcus pneumoniae. BL-S 578 was more resistant than cephalexin to the beta-lactamases produced by Klebsiella pneumoniae and Escherichia coli.


[Relations between the levels of neuroleptics and the doses, the therapeutic effects and the side effects]. The mass fragmentography and the gas-liquid chromatography are available actually for quantitation of very low levels of neuroleptics. The Radio-immunological assay is full of promise. Their plasma levels are not correlated with their dosage in a man to another, but, for some of them, there is lineary relation for one person. A correlation between their levels and their clinical efficacy is not certainly demonstrated for thioridazine and non transformed chlorpromazine; it could be possible for his sulphoxide metabolite and for butaperazine. The sides-effects are relatively correlated with their levels in the begining of the therepautic. The antiparkinsonian drugs, the antidepressants and the lithium could influenced them.


Neuropharmacological regulation of episodic growth hormone and prolactin secretion in the rat. The effects on GH and PRL secretion of several pharmacological agents known to modify central neurotransmitter action were determined in unanesthetized male rats. Phenoxybenzamine, an alpha-adrenergic blocker (5 mg/kg iv), abolished episodic GH secretion and caused elevation of serum PRL levels. Propranolol, a beta-adrenergic blocker (5 mg/kg iv), had no effect on GH secretion and caused a small but significant depression in PRL levels. Parachlorophenylalanine methyl ester, an inhibitor of tryptophan hydroxylase (300-350 mg/kg ip), resulted in significant inhibition of GH pulsatile secretion and suppressed PRL levels. Methysergide hydrogen maleinate (25 mg/kg iv), a serotonin receptor antagonist, also inhibited GH secretion, but produced a transient stimulation in PRL levels. Atropine sulfate (2 mg/kg iv) caused significant suppression in GH secretion, but had no effect on PRL. Picrotoxin, a gamma-aminobutyric acid antagonist, in a subconvulsive dose of 1-3 mg/kg iv, also depressed GH episodic secretion but did not affect PRL levels. These results indicate that several neurotransmitters, i.e., norepinephrine, serotonin, acetylcholine, and gamma-aminobutyric acid, found in high concentration in the hypothalamus, influence GH and PRL secretion.


Steroid metabolism in normal mammary gland and in the dimethylbenzanthracene-induced mammary tumor of rats. After incubation of [4-14C]progesterone with cell-free homogenates of 9,10-dimethyl-1,2-benzanthracene (DMBA)-induced mammary tumor of rats, 20 alpha-hydroxy-4-pregnen-3-one, 5 alpha-pregnane-3,20-dione, 20 alpha-hydroxy-5 alpha-pregnan-3-one, 3 alpha-hydroxy-5 alpha-pregnan-20-one, and 5 alpha-pregnane-3 alpha, 20 alpha-diol were identified as the metabolites. In normal mammary tissue, however, 4-pregnene-3 alpha-diol was isolated in addition to 5 alpha-reduced, and 3 alpha- and 20 alpha-hydroxy metabolites. When radioactive testosterone was employed as a substrate, 5 alpha-dihydrotestosterone and 5 alpha-androstane-3 alpha, 17 beta-diol were obtained as the metabolites of the mammary tumor. In the normal mammary gland, only 4-andorstene 3 alpha, 17 beta-diol was formed as its metabolite. Although the enzyme activities relevant to the metabolism varied among the tumor examined, the activity of 20 alpha-hydroxysteroid dehydrogenase in the mammary tumor was significantly lower than that in the normal mammary gland, whereas the activity of 5 alpha-reductase was higher in some of the mammary tumors than in the normal gland. The 5 alpha-reductase activity in the normal mammary gland was mostly localized in the crude microsomal fraction, whereas the same enzyme activity in the tumor was detected in all the organelle fractions. The activities of 20 alpha-hydroxysteroid dehydrogenase and NADPH-linked 3 alpha-hydroxysteroid dehydrogenase were found mainly in the cytosol fractions of the tumor and the normal tissue. The NADH-linked 3 alpha-hydroxysteroid dehydrogenase activity was detected only in the cytosol fraction of the normal mammary gland, but in the tumor studied, the activity of this enzyme was detected in all the subcellular fractions examined.


The effect of solubilization on the properties of the insulin receptor of human placental membranes. The insulin receptor for human placental membranes has been solubilized in Triton X-100 and its properties have been examined in detail. Binding of [125 I]iodoinsulin to the soluble receptor is markedly inhibited by increas-ng concentrations of Triton X-100, due to a fall in receptor affinity. In 0.02--0.10% Triton X-100, the soluble receptor exhibits all the essential characteristics of the intact or particulate receptor. These include strict specificity for insulin and its analogues, increase in steady state binding with decrease in temperature, a pH optimum at 7.8--8.0, and negatively cooperative site-site interactions. The initial association rate of [125 I]iodoinsulin and the soluble receptor is a direct function of temperature, but the level of steady-state binding is lower at higher temperatures due to a marked increase in dissociation rate. Scatchard binding plots are curvilinear and show a large increase in affinity at 4 C with no change in total binding capacity (R0); increased binding to the particulate placental membrane at 4 C is due chiefly to an increase in R3. Negative cooperatively in the soluble receptor has been confirmed by kinetic experiments; thus, the dissociation of [125I]iodoinsulin from the receptor in the presence of "infinite" dilution is accelerated in the presence of 10(-8) M unlabeled insulin. The apparent molecular weight of the placental receptor, determined by gel filtration on 6% agarose, is approximately 300,000. These studies show that the basic properties of the insulin receptor do not depend on it being an integral conponent of the cell membrane.


The possible roles of membrane organization in the activity of androgen biosynthetic enzymes associated with normal or tumorous mouse Leydig cell microsomes. The enzymes involved in conversion of pregnenolone to testosterone in Leydig cell tumors showed a wide distribution among smooth endoplasmic reticulum (SER), rough endoplasmic reticulum (RER), and cytosol, while these enzymatic activities in normal testes were associated primarily with smooth endoplasmic reticulum. Progesterone, used as a substrate in the presence of an NADPH-generating system, was metabolized to androstenedione and finally to testosterone by microsomes from some strains of tumor which did not form testosterone from exogenous labeled androstenedione. Treatment of microsomal membranes from normal testes with 0.1 M Ca++ and Mg++ caused a marked decrease in 17 beta-dehydrogenase activity, measured as conversion of exogenous [3H]androstenedione to [3H]-testosterone, without serious effects on activities of 3 beta-ol-dehydrogenase or 17 alpha-hydroxylase. Studies of initial velocity kinetics showed that treatment with magnesium ion resulted in a marked reduction in affinity of androstenedione for 17 beta-dehydrogenase while the maximum velocity was the same as in untreated microsomes. Also, experiments using [14C]progesterone and [3H]androstenedione simultaneously as substrates demonstrated that treatment with Mg++ ion made it more difficult for exogenous [3H]androstenedione to reach the active site of 17 beta-ol-dehydrogenase than [14C]androstenedione formed in the microsomal membrane from [14C]progesterone. Microsomal proteins were more easily solubilized and 3 beta-ol-dehydrogenase was more severely influenced by Mg++ ion in tumor membranes than in normal microsomes.


On the role of calcium in adrenocorticotropin-induced changes in mitochondrial pregnenolone synthesis. The importance of calcium in the ACTH-induced increase in adrenal mitochondrial pregnenolone synthesis was evaluated. In mitochondria prepared in the absence of EDTA and albumin, calcium enhanced the binding of cholesterol to cytochrome P-450 and subsequent pregnenolone synthesis. Although these effects of calcium were slightly greater in control than in ACTH-treated mitochondria, a sizeable effect of ACTH remained even at high calcium levels (500 micron). In mitochondria prepared from adrenals homogenized in fluid containing EDTA and albumin, ACTH-induced effects on pregnenolone synthesis were relatively poor unless calcium was added to the incubation mixture. High concentrations of added calcium (500 micron or greater) obviated the need for the labile protein required for ACTH-induced effects in intact mitochondria, presumably by disrupting mitochondria and allowing an "unrestrained" interaction of cholesterol with cytochrome P-450. Thus, cholesterol-rich mitochondria from ACTH plus cycloheximide-treated rats produced large amounts of pregnenolone when high (probably unphysiological) calcium concentrations were present. The present findings suggest that calcium is required at the mitochondrial level for ACTH-induced effects on pregnenolone synthesis, and the reported ACTH-induced increase in intraadrenal calcium may thus amplify the effects of ACTH on steroidogenesis. However, it seems unlikely that calcium is the agent primarily responsible for mediating the ACTH-induced steroidogenic effect at the mitochondrial level.


The charge heterogeneity of soluble human galactosyltransferases isolated from milk, amniotic fluid and malignant ascites. UDP-galactose: N-acetylglucosamine galactosyltransferase was isolated from pooled human milk, pooled amniotic fluid and from two different individual samples of malignant ascites. The purification procedure involving two successive affinity chromatography steps on N-acetylglucosamine--agarose and alpha-lactalbumin--agarose yielded an enzyme preparation homogeneous by size. Under non-denaturing conditions the ascites and amniotic fluid enzymes had identical electrophoretic mobility, but they moved faster than the milk enzyme. Isoelectric analysis in the presence and absence of urea resolved the milk enzyme into at least 13 different forms, nine of which had the same isoelectric points after refocusing. All enzyme forms showed similar activity when free N-acetylglucosamine, ovalbumin, sialic-acid-free ovine submaxillary mucin and glucose, in the presence of alpha-lactalbumin, were used as acceptor substrates. Comparative isoelectric focusing of the three galactosyltransferases revealed identical patterns of the amniotic and ascites enzymes, but only partial overlap with the milk enzyme, which was less negatively charged. Neuraminidase treatment of ascites and milk galactosyltransferases produced very similar focusing patterns. The possible structural basis for this charge heterogeneity is briefly discussed.


Yeast killer toxin: purification and characterisation of the protein toxin from Saccharomyces cerevisiae. Killer toxin from killer strains of Saccharomyces cerevisiae was isolated from concentrates of extracellular medium by precipitation in poly(ethylene glycol) and chromatography through glyceryl-controlled-pore glass. The toxin migrated as a single protein band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis. A molecular weight of 11470 was determined for the toxin protein from its electrophoretic mobility and amino acid composition. Gel filtration of the active toxin indicated that the 11,470-Mr monomer was the active unit. Electrophoretic comparison of extracellular concentrates from a killer strain and an isogenic non-killer showed the presence of the toxin protein only in the killer-derived material. The activity of the toxin was most stable between pH 4.2 and 4.6. At 30 degrees C toxin from a superkiller strain was more stable than that from a normal killer.


Determination of the interaction of ADP and dADP with copper(II), manganese(II) and lanthanide(III) ions by nuclear-magnetic-resonance spectroscopy. The aqueous solution conformation of the 1:1 complexes of ADP and dADP bound to a lanthanide ion have been determined by examination of the dipolar shifts and induced relaxation at pH 6.4. Apparent inconsistencies in the observed data are interpreted in terms of a gradually changing lanthanide-oxygen bond length from Pr3+ to Yb3+. The conformations of ADP and dADP are very similar showing an extended diphosphate, a 2E ribose conformation and with the adenine base displaying a small syn contribution. Relaxation data obtained from Mn(II) titrations are readily interpreted in terms of bidentate coordination to alpha and beta phosphates with the nucleotides retaining the same overall conformation found from the lanthanide study. No evidence to support an intramolecular water-bridged backbound structure is observed. The interaction with Cu(II) is more complex, coordination is observed not only at the diphosphate but also at two sites on the base, N-1, and the chelate formed between N-7 and the amino group. The relative importance of these sites is different for ADP and dADP and is also pH-dependent for ADP.


kappa-Carrageenase from Pseudomonas carrageenovora. A kappa-carrageenase was isolated from the cell-free medium of cultured Pseudomonas carrageenovora. From dodecylsulphate/polyacrylamide gel electrophoresis, a single protein (identified as the kappa-carrageenase) was detected in the medium. Activity against nominal carrageenan types and inspection of the products indicate the enzyme to be a kappa-carrageenase. Purification is described here by ammonium sulphate precipitation and subsequent CM-Sepharose CL-6B ion-exchange chromatography. Molecular weight was estimated as 35,000 by dodecylsulphate/polyacrylamide gel electrophoresis. Products of degradation were analysed by gel filtration, spectrophotometric assays and 13C nuclear magnetic resonance. These results are consistent with the product of limit digest being neocarrabiose 4-O-sulphate.


Pial arterial reactions to hyper- and hypocapnia: a dynamic experimental study in cats. In 29 cats, the extent and time-course of the pial arterial reactions to hypo- and hypercapnia were studied by means of the skull-window technique. The typical, well-known dilatations and constrictions during hyper- and hypocapnia were seen. The latent period for dilatation after the beginning of CO2-inhalation was ca. 20 sec. There was no stable relation observable between vessel diameter and arterial carbon dioxide tension (paCO2). Diameter changes lagged behind CO2-changes, indicating that CO2 acts via metabolic regulation, probably extracellular pH.


Relation between the test for gonadal stimulation with HCG and testicular histology in cryptorchism. A histological study, by means of bilateral biopsy, on 44 cryptorchid patients (26 of whom are monolateral and 18 bilateral), comparing them with the response obtained for the HCG gonadal stimulation test, is presented. A direct, although not always exact, relation is established between the testicular histology and the testosterone levels after the administration of 4,500 U of HCG.


Haemodynamic effects and pharmacokinetics of a new selective beta1-adrenoceptor agonist, prenalterol, and its interaction with metoprolol in man. The haemodynamic effects of the selective beta1-adrenoceptor agonist prenalterol were studied in healthy subjects before and after therapeutic doses of the selective beta1-adrenoceptor blocker metoprolol. Plasma levels of the drugs were also determined in order to calculate certain pharmacokinetic variables. Intravenous infusion of prenalterol 0.13, 0.25 and 0.50 mg induced a dose-dependent decrease in total electromechanical systole (QA2) and pre-ejection period (PEP).  The effect on left ventricular ejection time (LVET) was not significant. Increases in systolic blood pressure and heart rate were dose-dependent. Diastolic blood pressure did not change significantly. When metoprolol had been administered in a cumulative dose of 150 mg (mean maximal plasma level, 284 nmol/l) prenalterol had to be administered in doses that were twelve times higher than before the beta-blocker in order to induce the same haemodynamic effects. Prenalterol was rapidly distributed with an average half life of 8 min. This indicates that distribution equilibrium will be achieved within 30 min after intravenous administration. The overall elimination rate in the post-distributive phase corresponded to an average half life of 2.0 h.


Alterations in blood levels of carbohydrate and lipid metabolites and of cyclic AMP mediated by beta1- and beta2-adrenoceptors in beagle dogs: effects of procaterol, a new selective beta2-adrenoceptor agonist. The intravenous injection of isoprenaline (10 nmole/kg) into conscious beagle dogs caused significant increases in the blood level of lactate, glucose, FFA, insulin and cyclic AMP. These metabolic alterations induced by isoprenaline were blocked completely by pretreatment of the dog with propranolol (1 mg/kg). Butoxamine (10 mg/kg) antagonized isoprenaline-induced increases in glucose, lactate and insulin, but not the increases in FFA. Practolol (10 mg/kg) diminished the increase in blood FFA very strongly. Salbutamol, which is known to be an agonist of the beta 2-subtype in its bronchomotor and cardiovascular actions, produced marked increases in the blood concentrations of lactate, glucose and insulin but were without effect on the FFA level. Thus metabolic responses of conscious beagle dogs to beta-adrenoceptor agonists appeared to depend differentially on two types of beta-adrenoceptors: beta1-adrenoceptors are largely involved in lipolysis while beta2-adrenoceptors are involved in the regulation of blood glucose metabolism and insulin secretion. A new beta-adrenoceptor agonist, 5-(1-hydroxy-2-isopropylaminobutyl)-8-hydroxycarbostyril hydrochloride hemihydrate (Procaterol), was classified as a beta 2-agonist, because it markedly increased plasma concentrations of glucose, lactate and insulin but increased the plasma level of FFA to a lesser degree. The order of potency of beta2-agonists was procaterol greater than salbutamol greater than trimetoquinol. Metabolic responses of beagle dogs would be useful for appreciating the selectivity and potency of beta-adrenoceptor agonists and antagonists.


Isolated lung strips of guinea pigs: responses to beta-adrenergic agonists and antagonists. Isolated lung strips of guinea pigs were examined as an in vitro model for assessing the direct effect of beta-adrenergic drugs at the level of peripheral airways. Changes in intrinsic tone of thin strips of lung parenchyma were measured with an isometric force transducer. Isoproterenol, a nonselective beta-adrenergic agonist, and several beta-adrenergic agonists, soterenol, salbutamol, metaproterenol and ritodrine elicited a dose-related relaxation of lung strip. Responses to isoproterenol were antagonized by propranolol and the selective beta blocking agents butoxamine (beta2) and practolol (beta1). These results were compared to data obtained with the same compounds on isolated guinea pig atria. All agonists except ritodrine were full agonists in the lung strip whereas isoproterenol and metaproterenol were the only full agonists in the atrial preparation. In the atria, practolol was a more effective blocker of isoproterenol responses than butoxamine, and the reverse was true for the lung strip.


The action of verapamil on the rate of spontaneous release of transmitter at the frog neuromuscular junction. Verapamil is known to reduce Ca2+ entry in a variety of cells. At 10(-5) M it produces a small reduction in MEPP frequency at the frog neuromuscular junction, whereas the rate of spontaneous release rises following treatment at a concentration of 10(-4) M. This latter effect is augmented by raising [Ca2+]0 to 9 mM or, more especially, by raising the temperature from 17 to 23 degrees C. It is argued that both these opposing effects are related to the action of verapamil in modifying [Ca2+]i at the presynaptic terminals and it is suggested that the drug can affect both inward Ca2+ flux (so reducing the steady-state position of [Ca2+]i) and also, at higher concentration, either inhibit the membrane Ca2+ pump or cause the release of Ca2+ from intracellular Ca2+ stores (so raising [Ca2+]i).


Dopaminergic interaction with the respiratory control system in the rat. Apomorphine given to rats lightly anesthetized with halothane produces a dose dependent increase in respiratory frequency and minute ventilation. Although basal arterial CO2 tensions were not significantly altered by apomorphine, the mechanical response to exogenous CO2 exposure was greatly increased in rats given apomorphine. Haloperidol returned the apomorphine-stimulated respiratory pattern to control values. It is concluded that dopamine neurons may have important interactions with respiratory control.


Pre- and postsynaptic effects of alpha-agonists in the anococcygeus muscle of the pithed rat. In the pithed rat electrically induced contractions of the anococcygeus muscle were inhibited by clonidine (0.1--3.0 microgram/kg, i.v.), mecamylamine (1 mg/kg, i.v.) and guanethidine (1 mg/kg, i.v.). Only the inhibition produced by clonidine was antagonised by yohimbine (0.3 mg/kg, i.v.). At higher concentrations (10 microgram/kg, i.v.) clonidine increased the resting tension of the tissue; this effect was antagonised by phentolamine (1 mg/kg, i.v.). The anococcygeus muscle of the pithed rat was used to simultaneously assess pre- and postsynaptic alpha-adrenoceptor agonist activity. Guanfacin was more selective and tiamenidine less selective than clonidine for presynaptic alpha-adrenoceptors. Naphazoline and oxymetazoline were selective postsynaptic alpha-adrenoceptor agonists.


The effect of dopaminergic modifiers on morphine-induced analgesia and respiratory depression. The influences of the dopaminergic system on morphine-induced analgesia and respiratory depression were compared using modulators of dopaminergic activity. Blockade of dopaminergic receptors by haloperidol or pimozide produced a potentiation of morphine analgesia, while stimulation of dopaminergic activity by L-dopa methyl ester inhibited morphine analgesia. Morphine-induced depression of respiratory rate was potentiated by haloperidol and inhibited by pimozide or L-dopa methyl ester. These results suggest that the dopaminergic system plays a modulating role in morphine-induced analgesia, but not in morphine-induced respiratory depression.


Tumor acceptance modified by passage in hybrids with graft-versus-host reaction. A graft-versus-host reaction (GVHR) was produced in adult F1 hybrid mice by the injection of 10(8) parental strain spleen cells and 8 days later they were challenged with allogeneic third-party tumor. BALB/c Leydig cell tumor (C4092), C57BL/6 sarcoma (30795), and DBA/1 melanoma (S91) often grew progressively in B6D1F1, CD1F1, B6CF1 or their reciprocal hybrid recipients, respectively, when GVHR had been induced in these animals. Control, without GVHR, hybrids always rejected the tumor. The C4092 tumor was serially transplantable in untreated hybrids after its initial passage in unrelated GVHR-treated mice; the S91 grew in its first passage into untreated B6CF1 mice but thereafter was rejected by these hybrids; while the B6 tumor 30795 grew progressively only in the initial GVHR-treated CD1F1 or reciprocal hybrids. Reduced immunogenicity of tumors resulting from passage in unrelated recipients immunosuppressed in association with a GVHR is comparable to allograft adaptation achieved by such techniques as organ culture pretreatment and presents an additional method for attenuating rejection of allotransplants.


Experimental hypoplastic marrow failure in the mouse. In order to study the pathogenesis of aplastic anemia in man, hemopoietic stem cells were investigated in 'aplastic mice' the aplasia being induced by the immunological method. C3H/He (H-2k, Mlsc) received 600 rad whole body x-irradiation followed by the transplantation of 10(7) lymph node cells prepared from B10.BR mic e (H-2k, Mlsb). The C3H/He mice developed pancytopenia and marrow hypoplasia 21 days after these treatments. The total number of nucleated cells, CFU-S and CFU-C in the marrow and the wet weight and CFU-C of the spleen were markedly reduced. These findings are consistent with those of aplastic anemia in man and the model may provide a useful tool for the investigation of the pathogenesis of this anemia. Control mice that received irradiation only recovered from the damage 21 days later, while control mice that receive lymph node cells only showed no hematological changes.


A rotating bottle culture method with continuous replacement of the gas phase. A 'rotator' culture method is described which provides a continuous flow of oxygenating gas to cultures in rotating bottles. The system maintains constant O2 and CO2 levels in the culture medium throughout the incubation period. It also provides a more stable pH than systems with sealed culture bottles.


Brain microvessel hexokinase: kinetic properties. Kinetic differences between brain capillary and parenchymal hexokinase in the presence of glucose, ATP. fructose, potassium, sodium and different pH were established. Parenchymal hexokinase is more susceptible to glucose inhibition, can tolerate greater variations in the ATP concentration, is inhibited by increasing concentrations of fructose and potassium, and showed greater activity on the lower pH values. The data suggest that in brain parenchyma and endothelial cells of brain microvessels, there are 2 different enzymes with regard to the kinetics properties.


Evidence for a neuronal release of isotopically labelled gamma-amino-n-butyric acid (GABA) from the rat dorsal medulla in vivo. High potassium and electrical stimulation consistently increase efflux of labelled GABA from the in vivo superfused rat dorsal medulla in a calcium-dependent fashion. The depolarizing alkaloid, veratridine, also evokes a large increase in efflux of labelled GABA. These data strongly suggest release from a neurotransmitter pool in this region.


Inhibition of saline-induced diuresis in the rat by sulpiride. Sulpiride (120 mg/kg, i.p.) inhibited saline-induced diuresis in the rat, an effect not observed with haloperidol, clozapine, pimozide or chloromazine. The antidiuretic effect of sulpiride also occurred in hypophysectomized rats suggesting that the response was not prolactin-mediated.


[Tetrahydropyridoazepine and tetrahydropyridoazepinone derivatives. I. Derivatives of 2,3,4,5-tetrahydro-1H-pyrido[3,2b]azepine and of the corresponding lactam]. Preparation of N-derivatives of 2,3,4,5-tetrahydro-1H-pyrido[3,2-b]azepine and, in one case, of 2,3,4,5-tetrahydro-1H-pyrido[3,2-b]azepine-2-one was achieved by introducing the substituents COCl, CONH2, CO2(CH2)3N(CH3)2, COCH2Br and (CH2)3N(CH3)2. The pharmacological results indicate some effect on the ANS.


Molecular properties of the adrenergic alpha-receptor. III - Origin of topographical dualism in the reaction of a receptor thiol with symmetrical and unsymmetrical polyamine disulfides. Structure activity relationship studies previously led to the discovery of two distinct classes of tetramine-disulfides [(I) and (II)] each exhibiting optimum adrenergic alpha-blocking activity. The more potent one (I) was shown to uniquely discriminate between noradrenaline (NA) and adrenaline (A) binding sites. The unusual receptor topographical dualism toward (I) and (II) led to an investigation of the role of structural symmetry in their interaction with the receptor. To this end, a series of unsymmetrical disulfide analogs (x-z) where half of (I) was retained and the other half was made of unsubstituted N-(omega-aminoalkyl)cysteamines [as present in (II) and its homologs] were synthesized and evaluated as alpha-blockers. In addition, the role of the number of basic nitrogens on activity was also examined. It was found that the presence of four nitrogens is necessary for optimum activity. Moreover, it was discovered that optimum alpha-blocking potency is obtained when the respective halves of the two best prototypes (I) and (II) of symmetrical structures are fused [compound (XVI)]. This new unsymmetrical tetramine disulfide has a potency approaching that of (I) and its receptor saturation mechanism is similar. Moreover, it also shares with (I) the ability to discriminate between NA and A elicited responses. The effect of methylation of a single inner nitrogen on potency allowed the conclusion that each half of the unsymmetrical disulfide (XVI) respectively occupy half of the sites for (I) and half of those for (II). Accordingly, the previously observed topographical dualism toward (I) and (II) can best be accommodated by a model where two distinct sets of sites crossing each other over the same receptor thiol are involved. The possible significance of the anionic site multiplicity of the alpha-receptor is briefly discussed.


Molecular properties of the adrenergic alpha-receptor. I--Structural requirements for specific covalent occupancy by N,N'-bis--(5-aminopentyl)cystamine derivatives. The synthesis and alpha-adrenoreceptor blocking activity of several substituted analogs of the prototype alpha-blocker N,N'-bis-(5-aminopentyl)cystamine (APC) are described. The three optical forms of the analog carrying methyl groups on the carbons alpha- to the sulfurs were synthesized and shown to be equipotent and somewhat less active than APC. The omega, omega'-bis-guanidino analog of APC was less active. Significant improvement in potency was observed only with APC analogs carrying benzyl and substituted benzyl groups on the terminal nitrogens. Linking the terminal nitrogens of APC with a p.xyledenyl group so as to give the 26-membered analog caused a sharp drop in activity. The significance of these results as regards the alpha-receptor topography is discussed.


[Effect of neuroleptics on spontaneous and evoked noradrenaline release from brain nerve endings of rats]. Spectrofluorometric determination of the noradrenaline content in the non-purified fraction of the rat's brain stem synaptosomes and radioisotope method resulted in finding that carbidine is capable to release noradrenaline from the nerve endings. Azabutyron, droperidol and fluphenazine had no noticeable effect on this process. The release of noradrenaline with carbidine depends upon the presence of Ca2+ ions in the medium. This neuroleptic is also capable of intensifying the release of the mediator provoked by the potassium depolarization of the nerve endings.


[Antihistaminic and antiserotonin properties of new complex tropine esters]. The antihistaminic and antiserotonin properties of 16 new tropine esters, analogues of atropine, tropacin and tropaphen, were studied. All of them were found to lessen the spasmogenic effects of histamine and serotonin. The intensity of the antihistaminic and antiserotonin action of the drugs varied depending upon the structure of the radical at the alpha-carbon atom in the acidic part of the molecule. Both types of the activity are most marked in the desoxymethyl propyl and butyl analogues of atropine. The absence of the oxymethyl group at alpha-carbon in the series of atropine analogues is shown to facilitate the manifestation of the antihistaminic activity.


Initial experience with a new long-acting beta-blocker, nadolol, in hypertensive patients. Nadolol, a new beta-blocker that is not metabolized and has a uniquely long pharmacological half-life (up to 24 hours) was employed as the sole drug therapy for thirty-one patients with mild to moderately severe essential hypertension. Twenty-two patients had a reduction over 3 months of 10% or more in supine diastolic pressure, including twelve who became normotensive. Daily dosage, administered in two divided doses, ranged up to 640 mg; however, twenty patients required 160 mg or less. Twenty-three patients continued therapy for a median additional period of 15 months, during which satisfactory control was maintained in seventeen patients. Side-effects of nadolol caused termination of therapy in only one patient.


Angina pectoris: effective therapy once daily. Twenty-nine anginal patients were randomly allocated to nadolol once daily, or propranolol four times daily and titrated to optimum dosage over 14 weeks under double-blind conditions: all patients then continued on nadolol once daily. Efficacy was assessed by numbers of anginal attacks and nitroglycerin (GTN) usage, and exercise EKG on the bicycle ergometer. Results showed the two drugs to have similar effects regarding anginal attacks and GTN consumption, but nadolol produced better performance in exercise time. In the extended treatment period on nadolol, the improvements over baseline readings were maintained. No serious side-effects or laboratory abnormalities were encountered.


Takayasu's arteritis in Crohn's disease. This report describes a patient with coexistent Crohn's colitis and Takayasu's arteritis involving the thoracic aorta, superior mesenteric artery, and left hepatic artery. Unlike most previously reported cases of this association, characteristic histopathology is demonstrated. This case is also unusual in that the Takayasu's arteritis presented as aneurysmal disease instead of occlusive disease. We believe that Takayasu's arteritis may represent another of the many extra-intestinal manifestations of inflammatory bowel disease.


[Report on the round-table discussion of the subject "idea and experience" in connection with the 1977 annual meeting "process kinetics" of the German Academy of Natural Scientists Leopoldina (Halle/S., October 16, 1977)]. A report is given about a round-table-discussion upon the role and meaning of "idea and experience" in the creative scientific process. Notable representatives of mathematics, theoretical physics and geophysics, chemistry, theoretical and general experimental biology, and of medicine contribute in the discussion guided by C. F. V. WEIZSACKER to this theme from point of view of their disciplines. The components of meaning of "idea and experience" in their connection one to another may be paraphrased by such pairs of terms as "theory and practice", "theoretical or empirical", "law and appearance of a single phenomenon", "unity and diversity", abstract and concrete". It was demonstrated that in each of the mentioned scientific disciplines there is a natural, and that, starting from mathematics and going to biology and medicine, the weight in that relation will shift more and more to "experience". Many of the known methodological problems and difficulties will arise in the mentioned scientific branches if one stresses immoderately only one component of "idea and experience" by leaving the natural, discipline-related range of variation of the relation "idea and experience".


EcoRI activity: enzyme modification or activation of accompanying endonuclease? A study has been made of the factors and mechanism leading to appearance of the so-called EcoRI activity described by Polisky et al. (1975) in the restrictase EcoRI preparations. The preparations of purified restrictase EcoRI, precipitated at 0.9 ammonium sulphate saturation, as well as that obtained using standard techniques have been found to contain an admixture of an endonuclease which at neutral pH and high ionic strength multiply cleaves those DNAs which normally have only one recognition site for EcoRI. Under the standard conditions for EcoRI digestion this activity is found only when large amounts of freshly isolated enzyme are added to the incubation mixture and it is sharply enhanced by replacement of Mg2+ with Mn2+. The number and size of DNA fragments produced under such conditions practically do not differ from those found under the so-called EcoRI conditions, that is for alkaline pH values and low ionic strength. The optimum incubation mixture for the EcoRI activity has been found to be 10 mM Tris . HCl buffer (pH 8.8) + 2 mM Mn2+. Similar activity is induced also by addition to EcoRI solution of 40--50% glycerol or a number of organic solvents (dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulphoxide (DMSO), sulphalane (SP) in concentrations from 1 to 6%. The EcoRI activity induced by 50% glycerol or at alkaline pH values and low ionic strength is suppressed or sharply inhibited by 2--3 mM parachloromercuribenzoate (PCMB), while EcoRI is not sensitive to this agent. The DNA fragments cleaved by EcoRI have cohesive termini and can be easily ligated. It is suggested that the EcoRI activity can be due not only (or largely not) to modification of the "recognizing capacity" of the EcoRI restrictase but not activation of a latent specific endonuclease which is present in the restrictase preparation as an impurity.


[Absence of the inhibitor factor of the Mullerian structures. Report of a case]. A 30 year old patient with normal masculine phenotype, karyotype 46 XY with a congenital left inguinal hernia, was studied. This hernia contained (at surgery): uterus, tubes and a hypoplasic vagina, as well as testis. The Müllerian structures and the right testicle, were resected, and left orchidopexy was performed. FSH, LH, testosterone, prolactine, B-HCG in blood by radioimmunoanalysis, total estrogens, 17-ketosteroids and 17-hydroxicorticosteroids in urine, were all normal. Roentgenologic and endoscopic urologic studies were normal. A post-operative study of semen showed moderate oligospermia. The factors involved in masculine sexual differentiation, were reviewed, specially the inhibitory factor of Müllerian structures.


Effect of glucagon and pentagastrin on gastric mucosal potential difference in man. Glucagon given as an intravenous injection of 2 mg (0.57 mmol) or as constant intravenous infusion of 1.64 microgram/kg/h (0.47 nmol/kg/h) significantly increased gastric mucosal potential difference (PD) in man. Pentagastrin infusion of 2 microgram/kg/h (2.8 nmol/kg/h) dramatically reduced gastric PD in man. The effect of each of these hormones on PD was reversed by the administration of the other. Changes in PD induced by one hormone were not associated with reductions in blood levels of the other. There was an approximate correlation between changes in PD and pH of gastric aspirates, however, the patterns of PD and pH changes were at times dissimilar. This study indicates that administration of gastrointestinal hormones significantly alters gastric mucosal PD in man.


Comparative trial of sulphasalazine and oral sodium cromoglycate in the maintenance of remission in ulcerative colitis. Patients with ulcerative colitis in remission were randomly allocated to treatment with sulphasalazine (2 g/day) or oral sodium cromoglycate (160 mg/day or 2 g/day), and the relapse rates in these treatment groups were compared during continued treatment for one year. The percentage cumulative relapse rate after 12 months' treatment was 30% in the 33 patients treated with sulphasalazine compared with 71% in the 25 treated with high dose sodium cromoglycate, a highly significant difference (P less than 0.01). Patients allocated low dose sodium cromoglycate were only treated for a maximum of six months, and the relapse rate in these 12 patients was similar to that in patients on the high dose. These results suggest that oral sodium cromoglycate is considerably less effective than sulphasalazine in maintaining remission, and by analogy with results in other trials may be no more effective than placebo tablets.


Favourable effect of storage at 0 degrees C on the O2 transport function of heparinized blood. The biological value of heparinized blood was found to decrease less on storage at 0 degrees C than on storage at +4 degrees C. At 0 degrees C the P50 value decreases substantially later and less, the blood is less acidotic, and the UV absorption of the dialysate is less enhanced than at +4 degrees C. The findings justify a further search for the conditions suitable for the widespread practical application of storage at 0 degrees C.


[Alpha- and beta-adrenoceptor blocking actions of labetalol and effects on the myocardial function, coronary circulation and myocardial energy metabolism thereof (author's transl)]. In the isolated atria, trachea and aorta of the guinea pig and in the heart-lung preparation (HLP) of the dog, labetalol was 3.6 approximately 5.9 times less potent than phentolamine in blocking alpha-adrenoceptors, 1.4 approximately 3.6 times less potent as beta 1-blocker and 6 times less potent as beta 2-blocker than propranolol. Beta 1 and beta 2-blocking actions were 12 approximately 28 and 7 times more potent than alpha-blocking action, respectively. As regards beta 1-blockade, attenuation of the heart rate increase was more prominent than that of the augmentation of the contractile force. Both in the isolated atrial preparation of the guinea pig and in canine HLP, labetalol produced a positive inotropic and chronotropic effect and a slight increase in the coronary flow associated with a slight increase in the myocardial O2 consumption and improvement of the myocardial redox potential. Myocardial extraction and usage of lactate, pyruvate and glucose tended to be decreased and those of FFA decreased significantly. The positive inotropic and chrontropic effect was observed in HLP of 6-hydroxydopamine-pretreated dogs and was abolished with propranolol. Labetalol produced a relaxation of the isolated trachea of the guinea pig, which was inhibited with propranolol. In doses higher than those required for alpha- and beta-adrenoceptor blockade, labetalol produced calcium-antagonistic action.


Amylase activity of Torulopsis ingeniosa Di Menna. Torulopsis ingeniosa DI MENNA was found to possess an alpha-amylase strongly attached to the cell wall, its pH optimum being at 5.5, optimum temperature at 50 degrees C, highly sensitive to thermal inactivation. The enzyme was found to be induced by starch but the synthesis is not subject to a glucose effect.


Production of L-glutamic acid by a Bacillus sp. A strain of Bacillus cereus var. mycoides isolated from Burdwan soil produces L-glutamate in the medium. The strain is able to grow and produce in a synthetic medium but supplementation with casamino acid or yeast extract improves the yield. Maintenance of pH of the fermentation medium near neutrality prolongs the active growth period and improves the yield. Glucose and ammonium nitrate were found to be most suitable carbon and nitrogen sources, respectively. Cane sugar molasses (as a substitute for glucose) significantly stimulated the growth but glutamate production was less. Various B vitamins stimulate the growth and glutamate yield. The yield of glutamate under optimal condition is 5.2 g/l.


[New developments in the field of bronchospasmolytics]. Bronchial obstruction is mainly treated by bronchospasmolytics. They have different sites of action and work either by stimulating the beta-adrenergic receptors (beta-sympathicomimetics), by inhibiting the phosphodiesterase (theophylline-derivatives) or by blocking the cholinergic receptors (anticholinergica). Beta-sympathicomimetics do not only act via the beta-receptors but also by inhibiting the degranulation of the mast cells and thus preventing the liberation of spasmogenic substances. A very promising development is ipratropium bromide, an anticholinergic with a spasmolytic effect as pronounced as that of a beta2-adrenergic substance but with hardly any adverse side-effect. Glucocorticoids which are highly effective in bronchial asthma were shown to have also an "permissive effect" towards beta-sympathicomimetics.


[Rational therapy of peptic ulcers].

[Rational therapy of peptic ulcers]. Prerequisites of rational conservative treatment of peptic gastroduodenal ulceration are rational diagnostics(endoscopy, histology, cytology), present state knowledge of peptic ulcer pathogenesis, and knowledge of the drug to be used. Treatment should aim at: pain relief, promotion of ulcer healing, prophylaxis of ulcer relapse including consideration of side-effects and costs of the drug, which should be minimal. At present, rational recommendations can be given only for the treatment of the acute ulcer attack but not as to prophylaxis of recurrences. Patients with uncomplicated peptic ulceration should be treated as outpatients, whereas in case of complications (penetration, perforation, bleeding of ulcers, symptoms of stenosis, dramatic pain, or particularly adverse living conditions at home) patients need hospitalization.


[Drug interactions in the intestinal tract]. The mechanisms of drug-drug interactions are generally classified and interferences within the gastrointestinal tract are discussed. The latter mainly concern interactions in which absorption of a drug is affected. Unwanted reactions may easily be avoided by choosing carefully combination and time of administration of two or more drugs to be used in a patient.


[Testing antacid with intragastric Ph measurements and gastroscopy]. In connection with the testing of an antacid in form of a suspension, pH-measurement and gastroscopy are carried out in the same 11 patients. Essential criteria of the efficacy of an antacid are its buffering capacity and its retention in the stomach as a mechanical protective for the gastric mucosa. The Talcid suspension tested in this way showed a good period of stay in the stomach, often exceeding the period of acid-binding capacity. Compared with bicarbonate, its buffering period is about twice as long above pH 4.5 and about 2 1/2 times as long at lower pH's.


Effect of metabolic alkalosis on the B-cell sensitivity to alloxan in vivo. Metabolic alkalosis was induced in starved mice by treatment with NaHCO3, which did not significantly alter the blood glucose concentration, but protected against the diabetogenic effect of subsequently given alloxan. This protection and the alkalosis had disappeared 4 hr after NaHCO3 treatment. Protection against alloxan, and metabolic alkalosis, were found also in starved mice pretreated with sodium lactate. The findings indicate that metabolic alkalosis, directly or indirectly, protects against alloxan toxicity in vivo.


Comparison of two pig intestinal brush border peptidases with the corresponding renal enzymes. Intestinal dipeptidyl peptidase IV and gamma-glutamyltransferase were compared to the corresponding kidney enzymes with respect to immunological and electrophoretic properties. The influences of selected effectors on the two enzymes were also studied. The two kidney peptidases exhibited the reaction of total identity with the corresponding intestinal enzymes in immunodiffusion. Furthermore, the intestinal dipeptidyl peptidase IV and gamma-glutamyl transferase showed the same inhibition patterns as the corresponding kidney enzymes and the acceptor specificity of the intestinal gamma-glutamyl-transferase was found to be identical to that of the kidney enzyme. The electrophoretic mobilities of dipeptidyl peptidase IV from the two organs differed greatly. The difference was almost abolished by treatment with neuraminidase, suggesting that the variation in mobility was due to different contents of sialic acid. It is suggested that the intestinal brush border peptidases, dipeptidyl peptidase IV and gamma-glutamyltransferase, are closely related to the corresponding enzymes obtained from the kidney.


[Psychiatric intensive therapy after acute alkaloid withdrawal syndrome]. Two groups of patients (nine patients each) were treated for acute alcaloid-withdrawal-syndromes. One group was given peroral neuroleptic medication, the other was treated by neuroleptic cocktail administered via vena subclavia catheter. By statistic comparison of the two methods the superiority of the latter could be shown in that duration of withdrawal, the syndrome itself and hallucinatory transitional psychoses could be drastically reduced. A suitable prescription and dosages of the neuroleptic cocktail are presented. The results are compared to the ones in current literature.


Correlation between in vivo and in vitro functional tests for activated macrophages. Mice undergoing graft-versus-host reaction, skin grafting, and inoculation with tumor cells were tested for nonspecific resistance by intravenous challenge with Listeria monocytogenes. Peritoneal exudate macrophages from mice treated in a similar manner were tested in vitro for increased degradation of [1-14C]glucose, ability to degrade antigen/antibody complexes, ability to inhibit intracellular growth of listeria, and staining for beta-galactosidase. There was good correlation between in vivo resistance towards L. monocytogenes and in vitro inhibition of intracellular growth. There was also good correlation between increase in beta-galactosidase and in vivo resistance in mice undergoing a graft-versus-host-reaction.


Regulation of lactose catabolism in Streptococcus mutans: purification and regulatory properties of phospho-beta-galactosidase. Phospho-beta-galactosidase (P-beta-gal), the enzyme which catalyzes the first step in the metabolism of intracellular lactose phosphate, occurred at high specific activity in the cytoplasm in 12 of 13 strains of streptococcus mutans grown on lactose but not other carbon sources. The P-beta-gal from S. mutans SL1 was purified 13-fold using diethylaminoethyl-cellulose ion exchange and agarose A--0.5 M molecular exclusion column chromatography. The molecualr weight of the enzyme was estimated to be 40,000, and its pH optimum was 6.5 in three different buffer systems. P-beta-gal activity was inhibited by Co2+, Zn2+, and Cu2+, but other cations, ethylenediaminetetraacetic acid, orthophosphate, and fluoride had no effect upon enzyme activity. The kinetic response of P-beta-gal to a model substrate, o-nitrophenyl-beta-D-galactopyranoside-6-phosphate, obeyed Michaelis-Menten kinetics, and the Km for this substrate was 0.19 mM. In addition to being under genetic control, P-beta-gal activity was regulated by a number of biologically active metabolites. Enzyme activity was inhibited in a sigmoidal fashion by phosphoenolpyruvate. The M 0.5 V value for phosphoenolpyruvate was 2.8 mM, and the Hill coefficient (n) was 3. In addition, P-beta-gal exhibited strong inhibition by ATP, galactose-6-phosphate, and glucose-6-phosphate. In contrast to inhibition of P-beta-gal activity by phosphoenolpyruvate, the inhibition exerted by ATP, galactose-6-phosphate, and glucose-6-phosphate obeyed classical Michaelis-Menten kinetics; the Ki values for these inhibitors were 0.55, 1.6, and 4.0 mM, respectively.


Nutritional requirements for synthesis of heat-labile enterotoxin by enterotoxigenic strains of Escherichia coli. Optimal growth conditions have been established for production of heat-labile enterotoxin (LT) by both porcine and human strains of enterotoxigenic (ENT(+)) Escherichia coli. There were no unusual growth factor requirements, and some strains produced fairly high levels of LT in a basal salts medium containing 0.5% glucose if the pH was carefully controlled. Several amino acids markedly stimulated LT synthesis when added to the basal salts-glucose medium. Methionine and lysine were the most stimulatory for both human and porcine strains. Either aspartic acid or glutamic acid further enhanced LT synthesis in the presence of methionine and lysine, with aspartic acid being more stimulatory for porcine strains and glutamic acid more stimulatory for human strains. There were no apparent vitamin requirements and no unusual cations needed for toxin synthesis except that Fe(3+) was slightly stimulatory for porcine strains. The stimulation by Fe(3+) was observed only in the presence of the three amino acids, suggesting that the effect was indirect rather than on toxin synthesis. The carbon source also influenced the yield of LT. Glucose supported maximal synthesis, but other carbon sources which exhibit a high degree of catabolite repression also supported high levels of synthesis. Little or no LT was released below pH 7.0; therefore, because the pH drops during growth from 7.5 to 6.8, even in highly buffered media, it was necessary to adjust the pH to 8.0 to effect complete release of cell-associated toxin. The defined medium containing three amino acids reduced the amount of UV-absorbing material in culture supernatants about fivefold and increased LT activity for various strains from two- to fivefold over a complex Casamino Acids-yeast extract medium. Conditions found to be optimal for synthesis of LT were inhibitory for the heat-stable enterotoxin.


Effect of growth rate and glucose concentration on the activity of the phosphoenolpyruvate phosphotransferase system in Streptococcus mutans Ingbritt grown in continuous culture. Streptococcus mutans Ingbritt was grown anaerobically in a chemostat with a glucose limitation, as well as with an excess of glucose (amino acid limitation) at dilution rates (D) between 0.05 and 0.4 h(-1) (mean generation time = 12 to 1.5 h). The glucose-limited culture produced cells having 1.5- to 6.0-fold greater glycolytic activity than the cells from the glucose-excess culture. The preferred substrate for these cells was glucose, with the glycolytic rate for sucrose being only slightly lower; the rate for fructose was half that of glucose. The glycolytic rate of the glucose-limited cells was maximum at D = 0.1 h(-1), with a decline in rate as the growth rate approached D = 0.4 h(-1). A comparison of the activity of phosphoenolpyruvate phosphotransferase system (PTS) in the two types of cells showed that the glucose-limited cells had 1.7- to 5.6-fold greater PTS activity for the three sugars than the glucose-excess-grown cells. Whereas little difference was seen between the three sugars with the latter cells, the glucose-PTS had the greatest activity with glucose-limited cells, with the maximum in cells grown at D = 0.1 h(-1). Comparison of the rate of sugar uptake in the chemostat with the rate of PTS transport activity in the cells at each growth rate demonstrated that only under conditions of slow growth with a glucose limitation was the PTS system capable of supporting growth on glucose. Furthermore, PTS activity in cells grown with an excess of glucose was insignificant when compared with glucose uptake during growth in the chemostat. This evidence supports the observation that S. mutans possesses at least one other system, in addition to the PTS, for the transport of glucose into the cell. The organism was, however, devoid of glucose-proton symport transport activity.


Longitudinal study of experimentally induced periodontal disease in Macaca arctoides: relationship between microflora and alveolar bone loss. Macaca arctoides monkeys develop periodontal disease, and they harbor a periodontopathic indigenous flora largely similar to that of humans. This study showed that various Haemophilus isolates and H2O2-splitting asaccharolytic Bacteroides melaninogenicus strains constituted major segments of the monkey periodontal microflora. These organisms have not been previously identified among human isolates. Furthermore, the present data revealed that asaccharolytic B. melaninogenicus strains increased in proportion from a few percent to about 66% of the total isolates concomitant with the development of a significant loss of alveolar bone mass. Hence, this study strongly implicates B. melaninogenicus subsp. asaccharolyticus and closely related strains as important pathogens in actively destructive periodontal disease.


Cell envelope of Neisseria gonorrhoeae CS7: peptidoglycan protein complex. Treatment of cells grown to exponential phase with 4% sodium dodecyl sulfate for 3 h at 100 degrees C resulted in solubilization of all cellular components except for peptidoglycan. In most strains, cells cultured in liquid gonococcal broth at pH 7.2 yielded a peptidoglycan composed primarily of N-acetylmuramic acid N-acetylglucosamine, alanine, glutamic acid, and diaminopimelic acid in a molar ratio of 1:1:2:1:1. The peptidoglycan in these cells accounted for 1 to 2% (dry weight) of the cells. However, in cells cultured at pH 6.0, the dry weight of peptidoglycan increased to 4 to 13%. Preliminary investigations indicated that the apparent increase in weight is strain dependent and is due in part to associated protein(s). Neisseria gonorrhoeae strain CS7 had elevated amounts of protein associated with the peptidoglycan regardless of growth pH. The peptidoglycan-protein complex could not be dissociated by additional extraction with sodium dodecyl sulfate, 10 M LiCl2, or ethylenediaminetetraacetate or by 7.5% polyacrylamide gel electrophoresis. The complex could be degraded by lysozyme, trypsin, chymotrypsin, Pronase B, and Chalaropsis sp. muramidase.


Pneumococcal adherence to human epithelial cells. Evidence is presented that pneumococci adhere poorly to oropharyngeal cells in vitro and that the capsule may interfere with adherence. A brief survey indicated that pneumococci may also adhere poorly in vivo.


Inhibition of the interaction between the complement component Clq and immune complexes. Several groups of compounds were examined for their ability to inhibit in vitro the binding of Clq to insoluble immune complexes. As expected from previous studies, polyinosinic acid, liquoid, sodium pentosan polysulfate, aliphatic diamines and heparin are good inhibitors. This study shows that compounds with much lower toxicity, such as certain amino acids and substances with vitamin B6 activity (pyridoxal-5-phosphate:P5P) were also capable of decreasing the binding of Clq. Using methods of equilibrium dialysis and difference spectra, it was shown that this compound binds to both, Clq und IgG antibody by forming Schiff bases. Clq binds approximately 10 times more P5P when compared to IgG. Immune complexes prepared with IgG antibody modified by P5P and stabilized with sodium borohydride had the same complement-fixing and Clq-binding capacity as normal immune complexes. This suggests that the inhibition of Clq-binding to immune complexes by P5P is due to a modification of lysyl resides in Clq. Collagen and IgG fragments derived from Fc were also found to inhibit Clq binding to immune complexes, but at higher concentrations than the above small-molecular compounds and with a different mode of action.


Clinical and laboratory findings on admission to an alcohol detoxification service. A review of 260 consecutive admissions for alcoholic withdrawal to the Alcohol Detoxification Service of the Los Angeles County--University of Southern California Medical Center shows that 98% of the patients fulfill the criteria for alcoholism as stated by the National Council of Alcoholism Criteria Committee. Previous alcoholic history, current symptoms, physical findings, and laboratory tests indicate frequent multiple organ dysfunction in acute alcohol withdrawal. Hallucinations occurrred in more than 46% of patients with a blood alcohol over 100 mg%. More than 90% of the patients had one or more abnormal laboratory values, and the serum gamma glutamyl transpeptidase was the most commonly elevated laboratory test.


Recent advances in diagnosis and management of galactorrhea. Using prolactin suppression (L-dopa) and provocative (thorazine) tests along with the clomid test the intactness of hypothalamico-pituitary axis was assessed in a group of eight patients  with galactorrhea syndrome. Pituitary neoplasm was suspected in three cases on the basis of elevated plasma prolactin and its abnormal response to L-dopa, thorazine, and clomid tests. Trans-sphenoidal exploration of the pituitary gland in these three cases confirmed the diagnosis. In these cases, however, the commonly used parameters for evaluation of the pituitary gland (skull X ray, visual field measurement, echoencephalogram, etc.) failed to reveal the presence of the pituitary lesions. In two of the three cases, normal menses recurred following extirpation of the tumors and conception was achieved successfully. In the remaining patient the operation failed to remove the entire pituitary lesion and it was necessary to irradiate the pituitary gland as the patient continued to have persistent galactorrhea and hyperprolactinemia. The importance of the prolactin assay, and its response to stimulation and suppression tests for proper evaluation of galactorrhea was fully discussed.


Treatment of infertility caused by antisperm antibodies. Twenty infertile couples with antisperm antibodies in the male or in the female partner were scheduled for treatment. In 15 couples the male and in 5 couples the female partner was the antisperm antibody carrier. In all the couples the result of the in vivo and in vitro sperm-penetration test was negative or poor. The SCMC-test was strongly positive in each of the couples. In all the couples IgG and IgA antisperm antibodies could be demonstrated on the spermatozoa or in the cervical mucus. It was postulated that antisperm IgA and not antisperm IgG is responsible for the penetration reduction of spermatozoa in cervical mucus and for the "shaking phenomenon" in the SCMC-test. Intrauterine inseminations, performed in 20 couples, resulted in four pregnancies. Condom therapy in three couples, for at lest 6 months, had no result. Two men were treated with 96 mg methylprednisolone per day for 7 days; this resulted in a slight decrease of the sperm-agglutination titre, but no pregnancy occurred.


The "unexplained" poor postcoital test.

The "unexplained" poor postcoital test. In 30 or 32 infertile couples with an unexplained negative or bad in vivo and in vitro sperm penetration test, we obtained a strongly positive Sperm Cervical Mucus Contact test (SCMC-test) and demonstrated the presence of antisperm antibodies in the male or female partner. In these 30 couples 25 of the male partners had a sperm-agglutination titre of at least 32 in the serum and at least 4 in the seminal plasma. In the five remaining couples the female partner showed a minimum sperm-agglutination titre of 16 in the serum and a cervical mucus titre of at least 128. In 48 couples with a fair or good sperm penetration in cervical mucus, in vivo and in vitro, we never found a strongly positive SCMC-test. In 43 of these couples the SCMC-test was negative. Only one man in the latter group had sperm-agglutinating activity in the semen. In a group 32 couples, with a negative SCMC-test, there was no or only weak sperm-agglutinating activity in the cervical mucus, although 2 women had moderate sperm-agglutinating activity in the blood serum. Based on these data we conclude that the so called "unexplained" poor postcoital test is almost always due to the presence of antisperm antibodies in the semen or in the cervical mucus. We consider the SCMC-test not only to be a simple and reliable technique for detecting the presence of these antisperm antibodies, but also a method of demonstrating the mechanism by which antisperm antibodies decrease the chance of conception.


A quantitative study of the effects of acetylsalicylic acid on spermatogenesis and organs of the rat. Although the occurrence of prostaglandins in the male reproductive organs is consistent, their physiological role in fertility and reproduction is not known. The influence of acetylsalicylic acid, an inhibitor of prostaglandin synthesis, on the male reproductive tract was investigated in Sprague-Dawley rats of proven fertility. Acetylsalicylic acid dissolved in phosphate buffer was administered once a day at two dosages (300 mg/kg body weight and 150 mg/kg body weight) over a period of 12 days and a period of 6 days. The animals were killed 24 hours after the final treatment and the testes, epididymides, ductus deferens, seminal vesicles, kidneys, and adrenal glands were removed and placed in Bouin's solution. Subsequently, the tissues were cleaned and weighed and the testes were prepared for quantitative study under the light microscope. Organ weights were not significantly altered in the animals that were treated with acetylsalicylic acid. Cell counts indicated that there was a significant increase in the mean number of preleptotene spermatocytes and spermatids in those animals treated with the drug for 6 days at a dose of 150 mg/kg body weight. Treatment at the same dosage for a period of 12 days produced a significant decrease in the mean numbers of preleptotene and pachytene spermatocytes and spermatids. The mean diameter of the seminiferous tubules was also significantly decreased in the latter group of animals. In the group of animals treated with ASA for 12 days at a dose of 300 mg/kg body weight the mean diameter of the seminiferous tubules was significantly increased. No clear conclusion as to the effect of the drug on spermatogenesis or the various organs could be drawn.


Significance of sperm antibodies in human fertility. Indirect immunofluorescent, macroagglutination and immobilizing sperm antibody estimations were performed as part of a prospective study on a representative group of 63 infertile couples. Serum from both, cervical mucus and seminal plasma were tested and the clinical use of the tests evaluated by considering the pregnancies that occurred and the causes of infertility that were discovered in the subsequent year. The indirect immunofluorescent test had the largest number of positives although more were found in seminal plasma than elsewhere. In these, both male and cervical factors were implicated and in this media there were less positives pregnant with the acrosome and midpiece patterns absent. The macroagglutination test had no positives pregnant and again more significance was attached to positives in the male. Similarly, immobilization results showed most positives in seminal plasma where the male and cervical factors featured prominently. It would, therefore, appear that if any clinical use is to be derived from the three tests in the search for antisperm antibodies as a cause of infertility, testing the male and especially seminal plasma appears to correlate best. If treatment is to be recommended, therefore, it should be concentrated on the man.


Correlation between in vivo inhibition of gonadotropin release induced by LH-RH and the blockade of ovulation by synthetic analogues of LH-RH. Several antagonists of LH-RH were examined for their anti-LH/FSH releasing activities in an immature male rat assay and for their ability to block ovulation. The in vivo inhibition of the release of LH and FSH induced by LH-RH over a 4-h period of time and the antiovulatory activity of these analogues were parallel to each other. The analogues that exhibited at least a 95% inhibition of LH and 77% of FSH release at 1 h and 75% and 40% of FSH release at 4 h in immature male rats, gave high blockade of ovulation. [D-Phe2, D-Trp3, D-Phe6]-LH-RH was the most potent analogue of the series and it seems to be active in men.


The correlation between sperm count and testicular biopsy using a new scoring system. Biopsies of 320 testes were evaluated and a new scoring system for describing testicular biopsies was used. This scoring system uses four criteria in the evaluation of the biopsy: the diameter of the tubules (D), the thickness of the peritubular membrane (M), the population of cells within the tubules (P), and the spermatogenic maturation degree (S). The score of each biopsy was correlated to the sperm count of each patient. A typical correlation between the biopsy score and the sperm count was found in cases with sperm count between 0 and 12 million/cc. The clinical value of the new scoring system in describing typical clinical syndromes and in the evaluation of the prognosis of treatment was emphasized.


Delayed diagnosis in a case of secondary amenorrhea caused by a long arm isochromosome--X; i(xq). Secondary amenorrhea was observed in a 27-year-old woman with a structurally abnormal isochromosome--X, i(Xq), in all cells analyzed. The apparent phenotypical variability that may be observed in subjects with functional monosomy for the short arm of X, must be taken into consideration when patients with amenorrhea are clinically investigated. A correct, and early diagnosis of the etiology of the dysgenetic state of the gonads may only be obtained by accurate cytogenetic investigations, i.e., by utilizing various modern banding techniques.


Clinicopathologic implications and diagnosis of nongranulomatous nonspecific endometritis. The analysis of 600 subsequent endometrial biopsies from urban gynecologic outpatient clinics was carried out with special regard to nongranulomatous and nonspecific endometritis. The total incidence of inflammatory changes reached 10.8%; after excluding all postabortion and puerperal endometria 9.4% of the curettings showed endometritis. The relative occurrence of acute, subacute, and chronic disease was respectively 6.2, 30.8, and 63.0%. The etiologic factor remained obscure in 64.7% of all the cases. The influence of inflammatory changes on the hormonally induced modifications of the endometrium are discussed.


The separation of peptide hormone diastereoisomers by reverse phase high pressure liquid chromatography. Factors affecting separation of oxytocin and its diastereoisomers--structural implications. Experimental conditions and parameters involved in high performance liquid chromatography (HPLC) separations of the peptide hormone oxytocin and seven of its diastereoisomers, namely [1-hemi-D-cystine]-, [2-D-tyrosine]-, [4-D-glutamine]-, [5-D-asparagine]-, [6-hemi-D-cystine-], [7-D-proline]-, and [8-D-leucine]-oxytocin, on reverse phase columns were investigated. The effects of solvent, pH, and salt concentration were studied. Using the solvent systems 10% tetrahydrofuran-ammonium acetate buffer or 18% acetonitrile-ammonium acetate buffer and the muBondapak C18 support, oxytocin was separated from each of its diastereoisomers under all conditions studied, but the order of elution of diastereoisomers was highly dependent on solvent and to a lesser extent on pH. Separations of the hormone and its diastereoisomers on reverse phase HPLC and on classical partition chromatography on Sephadex G-25 were compared. The results are discussed in terms of the interactions of the solute with the reverse phase column and the solvent system. Implications of these findings in terms of the different solution conformations of the peptides are discussed.


Haloacetyl groups as reversible protection of the amino function: cleavage with 2-aminothiophenol. Haloacetylamino acids and haloacetyl peptides react rapidly with 2-aminothiophenol in weakly alkaline media to yield 2-aminothiophenoxyacetyl derivatives. These intermediates are subject to acidolysis under mild conditions with release of free amino acids or peptides. With this mild method for removal of the haloacetyl group N-haloacetoxysuccinimide derivatives, which rapidly and specifically acylate amino groups of polypeptides in aqueous solutions, become promising reagents for the reversible protection of amino groups. The chloroacetylation of amino groups in lima bean trypsin inhibitor and the quantitative removal of the chloroacetyl groups demonstrate the applicability of the method for polypeptides. The haloacetyl group also serves an analytical function in that treatment of a completely or partially haloacetylated polypeptide with cysteine forms one carboxymethylcysteine residue per haloacetyl group in the polypeptide derivative. Carboxymethylcysteine is readily measured by amino acid analysis of acid hydrolysates. Approaches to further improvement of conditions for removal of haloacetyl groups are discussed and potential applications of the general chemistry of 2-haloacids to modern polypeptide chemistry are outlined.