Inhibition by alpha-adrenoceptor agonists of renin release in vitro. A variety of alpha-adrenergic agonists encompassing a broad range of concentrations were used to investigate the existence and nature of a putative alpha-adrenergic mechanism inhibitory to renin release, which may operate at the level of the juxtaglomerular apparatus. For this purpose rat renal cortical tissue incubated in vitro was used. Concentrations of noradrenaline, adrenaline and methoxamine of 10(-6), 10(-5), 10(-4) and 10(-3) M caused significant dose-related inhibition of renin release. The inhibition of release by these doses was reversed completely by 10(-4) M phentolamine. In contrast, phenylephrine, oxymetazoline and clonidine did not inhibit renin release. The results support the concept of an alpha-adrenergic mechanism inhibitory to renin release and show that high concentrations of alpha-adrenergic agonist are required for its operation in vitro. The manner in which this inhibitory mechanism affects renin release under physiological circumstances remains to be demonstrated.


Dopamine agonist-induced hyperglycemia in rats: effects of lergotrile mesylate. Lergotrile and apomorphine, two direct-acting dopamine agonists, caused marked hyperglycemia in fasted rats, while compounds which release endogenous dopamine (amphetamine, methylphenidate) or inhibit dopamine reuptake (LR5182), failed to elevate blood glucose. The effect of lergotrile was dose dependent, causing blood glucose to rise 3-fold above resting levels at 5 mg/kg (i.p.). Blood glucose increased prior to the onset of the behavioral signs of dopamine stimulation. The effect of lergotrile was attenuated by phentolamine, propranolol or butaclamol. Adrenalectomy also prevented lergotrile-induced hyperglycemia. These data indicate that hyperglycemia is not a property of all compounds that cause dopaminergic effects. The action of lergotrile might be indirect, perhaps mediated through release of catecholamines from the adrenal glands.


Amphotericin B induced structural changes of the erythrocyte membrane. Incubation of human erythrocytes in isotonic NaCl-sucrose medium with a relative high concentration of amphotericin B results in the occurrence of distinct structures on the membrane faces as visible by freeze-fracturing. Such kind of structures is known to be formed by phase separation of the lipid components (domain formation). The induction of phase separation is considered to be an additional effect of amphotericin B.


Direct measurement of the pH in the stomach of the conscious rat, using a special electrode. The construction and use of a specially designed electrode to measure gastric pH in the conscious rat is described. Measurement of gastric hydrogen ion activity with this device is rapid and reproducible in starved rats. A dose-related increase of gastric pH was obtained after s.c. injection of the anticholinergic isopropamide.


Recognition and regulation of midwives.

Recognition and regulation of midwives. There is an urgent need for internaltion standards for the recognition, training and licensure of midwives. It is important for obstetricians to use their influence constructively to help meet the need.


The midwife as a counselor on nutrition. Amid divided opinion among professionals about the effect of nutrition on the outcome of pregnancy, one indisputable fact which emerges is that a good diet is essential during pregnancy and lactation. The role of the midwife in counseling and educating pregnant women on their own and their babies' nutritional need is discussed.


The role of traditional birth attendants in family planning programs in Southeast Asia. The training and utilization of traditional birth attendants (TBAs) in maternal and child health and family planning programs in Indonesia, the Philippines, Thailand and Malaysia are discussed. Special efforts to organize and train TBAs for family planning in Malaysia are examined in detail. Import factors for successful utilization of TBAs include: (a) definite assignment of functions and tasks, (b) organization of good operational steps and (c) implementation of good supervisory activities.


Progress and problems in the care of mother and baby. The expanded role of the midwife as accepted practice is cited as an element contributing to improvement. A plea is made for a simplified system of recording perinatal events which would lead to a better study of progress and problems in the care of mother and baby.


The midwife and the family unit.

The midwife and the family unit. The activities of the midwife dealing with home deliveries or working in a ward and her relationship with pregnant women are discussed.


The midwife in Indonesia.

The midwife in Indonesia. In Indonesia, where a large percentage of the population lives in rural areas, the traditional midwife or dukun is essential in the field of obstetrics. Means to further her knowledge and improve her skills so that the population at large may benefit are discussed.


The use of paramedics in family planning services in Iran. In the Family Planning Department of Farah Maternity Hospital, from 1968 to 1976, paramedics were trained to administer all reversible methods of family planning and pregnancy termination up to ten weeks and to perform a limited number of abdominal and vaginal tubectomies (1). From 1974 to 1976, all the above methods, except tubal ligation, were performed satisfactorily on 460 417 acceptors at a clinic not located in the hospital. The complication rate of 1%-5% was the same whether the procedures were performed by paramedics or by physicians. The project has been economical. It has been accepted by both patients and paramedics and has freed doctors to carry out other work.


Genital tumor as a presenting symptom of renal hypernephroma: a case report. A case of hypernephroma is presented in which the initial finding was heavy bleeding from a vaginal tumor. Fewer than 90 cases of genital metastases in males or females have previously been reported.


Reversal of vasectomy.

Reversal of vasectomy. Vasovasostomy to reverse a previous vasectomy for sterilization was attempted for 27 men, the procedure being technically impossible in only one case. A testicular biopsy was performed at the time of operation and a number were investigated for cell-mediated immunity to sperm and for the presence of circulating sperm-agglutinating and cytotoxic antibodies. The first 17 cases have been studied and of these there have been 11 pregnancies, ten of which have already come to term with the birth of normal infants, including one set of twins. Of the rest, two are known to have oligozoospermia and four have been lost to follow-up, although two of them were euspermic when last examined. In spite of these encouraging results, it is considered that there are no grounds for alterning the present basis of vasectomy counseling which is that the operation is likely to be irreversible.


Serum cholesterol and hemoglobin levels and calorie requirements in pregnant Nigerian women. A study conducted on 50 healthy senior staff wives (socioeconomic group 1) and 63 healthy junior staff wives (socioeconomic group 2) attending the antenatal clinic at the University of Ife Health Center showed marked differences in the level of serum cholesterol and hemoglobin between the two groups. It was also found that women in group 2 took 6.8% of their total calorie requirements as fat and group 1 women took 12.0% of their total calorie requirements as fat.


Maternal mortality in an Israeli hospital: a review of 23 years. In spite of great strides in obstetrics, maternal mortality has been completely eliminated. Possible changes in the causes of mortality are examined for three periods of time (1954-1961, 1962-1971 and 1972-1976). The overall incidence was 3.6/10 000, changing through the three periods from 4.9 to 4.3 and finally to 3.0/10 000. Vascular accidents were the cause of death in almost one third of the cases, emerging as the most important etiologic factor. Older age and higher parity did not seem to influence the incidence of obstetric deaths. Cesarean section was involved in ten of 23 cases in which the death was directly related to the pregnancy and delivery. In six patients there was a rupture of the uterus. The number of preventable deaths has decreased steady, but research into the problem of vascular accidents and dampening of the enthusiasm for cesarean sections may further improve the situation.


Loss of beat-to-beat variability and a negative oxytocin challenge test: an ominous prognostic sign. The reputation of the predictive accuracy of a negative oxytocin challenge test (OCT) has been somewhat tarnished by recent sporadic reports of intrauterine fetal death relatively soon after a negative OCT. We have analyzed probable causes and the possibilities of reducing to a minimum "false-negative" results of the OCT. In particular, several of these reports did not take into account the loss of baseline fetal heart rate (FHR) variability recorded during the OCT and, in the absence of late decelerations, the OCT was interpreted as negative. We suggest that recordings showing a loss of baseline beat-to-beat FHR variability and a negative OCT illustrate a complete inability of the fetus to react to any stimulus and that, in these cases, a negative OCT should in no way be reassuring, but rather a warning sign of severe fetal compromise. Two cases are presented to illustrate this phenomenon.


The endodermal sinus tumor: special features. Eight cases of endermal sinus tumor, two extragonadal and six gonadal, seen at the American University of Beirut are reviewed. Three of them present special clinical and pathologic features that further substantiate the extraembryonic origin of this teratoid tumor and its association with dysgenetic gonads. We consider the two histologic types, endodermal sinus pattern and polyvesicular vitelline architecture, to be part of the same tumor. The prognosis in this series was unfavorable.


Fetal absorption of 3H-leucine injected into the amniotic sacs of pregnants rats. When injected into the amniotic sacs of fetal rats in late pregnancy, 3H-leucine is rapidly absorbed and incorporated into fetal proteins as determined by liquid scintillation spectrometry. Peak values in the fetal liver and pancreas occur around four hours after injection. Approximately 21% of the radioisotope is retained in the injected fetus at 24 hours. The remainder escapes into the maternal circulation and accounts for a gradual increase in levels in noninjected fetuses from the same litter and for persistently high counts found in maternal serum as late as 24 hours after the injection. Specific activity of the radioisotope in fetal tissues is considerably higher (100- to 200-fold) when 3H-leucine is injected inth the amniotic sac rather than intravenously into the mother. The possible treatment of various fetal disorders by means of intra-amniotic injections is discussed.


Birth weight, gestational age and other indices of fetal maturity. A number of indices for evaluating the maturity of the newborn were studied. Indices of anthropometric, anatomic, reflex and physiologic development were examined to evaluate their association with birth weight and gestational age; the validity of these diagnostic indices is discussed. Studies are recommended to establish a maturity score to identify high-risk infants at birth.


Correlation of ominous fetal heart rate pattern and scalp blood pH with one-minute Apgar score. Fetal minitoring data were analyzed in 176 labors that developed ominous fetal heart rate deceleration patterns (FHRDP) during the two hours prior to delivery in an effort to identify how neonatal condition could be most accurately predicted. It was found that ominous FHRDP corresponded to a one-minute Apgar score of 6 or less in only 15.3% of cases. When patterns with fetal scalp blood pHs (FSBpH) of less than or equal to 7.20 (n = 18) were considered, 44.4% had an Apgar score of 6 or less. Mean Apgar scores for those neonates who had a pH of less than or equal to 7.20 were significantly less than the mean Apgar score of the control group (p less than 0.001 for all deceleration types). The result reveals that FSBpH has a better predictive value than FHRDP alone for neonatal depression. It is suggested that, as soon as ominous FHRDP occurs during labor, FSBpH be performed to assess not only fetal status but also to predict neonatal outcome.


Current status of immunologic pregnancy tests. Information about the mechanism of various pregnancy tests is summarized. The sensitivity of these tests as well as their advantages and limitations in a clinical setting are presented and evaluated in an effort to maximize their usefulness in a clinical setting.


Recessive congenital myotonia and pregnancy. There is a group of genetic disorders of muscle associated with myotonia. Congenital myotonia, usually a benign disorder and myotonic dystrophy, potentially a serious disorder, belong to this group. Uterine recordings during labor in patients with congenital myotonia have not been previously reported. These records have been abnormal in patients with myotonic dystrophy which has been associated with serious obstetric complications. A case of congenital myotonia in pregnancy is reported. The record of uterine contractions was normal. Although the patient delivered a stillborn male, the etiology of the intrauterine fetal demise is obscure.


The occurrence of a distinct high molecular weight form of serum non-suppressible insulin-like activity. Gel filtration of acromegalic or normal serum at acid pH gave two distinct species of non-suppressible insulin-like activity (NSILA), one of high MW and the other of low MW (approximately 7000 daltons). The acid-stable high MW form remained high MW on rechromatography in acid. Gel filtration of serum at neutral pH however, gave only high MW activity, which remained high MW when rechromatographed under neutral conditions but split into both high and low MW forms when rechromatographed in acid. These results indicate that there are at least two circulating forms of NSILA--a low MW form which circulates in serum bound to a carrier protein in an acid-labile high MW complex and a species which circulates only as a stable, discrete high MW protein.


Reduction of cinerulose in aclacinomycin-A by soluble and microsomal cinerulose reductases. The in vitro metabolism of the antitumor anthracycline antibiotic, aclacinomycin-A, was studied using rat liver homogenate. In the presence of NADH or NADPH, aclacinomycin-A was converted to aclacinomycin-A analogs, MA144 M1 and MA144 N1, which were stereospecifically reduced at the keto group of the C-4''' position of L-cinerulose in aclacinomycin-A. Subcellular fractionation indicated that the production of MA144 M1, which was reduced to L-amicetose, was catalyzed by NADPH-dependent soluble cinerulose reductase I, and the production of MA144 N1, which was reduced to L-rhodinose, was catalyzed by NADPH-dependent soluble cinerulose reductase II and NADH-dependent microsomal cinerulose reductase. The properties of these three enzymes were studied. Soluble cinerulose reductase I which produces MA144 M1 showed a optimum pH at 6.3, Km values of 3.3 x 10(-4) M for aclacinomycin-A and 3.2 x 10(-5) M for NADPH. Soluble cinerulose reductase II which produces MA144 N1 showed a pH optimum at 6.3 and Km values of 2.0 x 10(-3) M for aclacinomycin-A and 4.0 x 10(-5) M for NADPH. All thesse reductases were sensitive to sulfhydryl reagents and were inhibited by vitamin K3. Microsomal cinerulose reductase showed sensitivity to diconmarol and ferrous ion. The main nondegradative pathways of aclacinomycin-A were discussed from these results.


Reduction of anthracycline glycoside by NADPH--cytochrome P-450 reductase. The in vitro degradation of the new antitumor anthracycline antibiotic, aclacinomycin-A, was studied using rat liver homogenate. In the presence of NADH or NADPH, the glycosidic bond at C-7 position of aclacinomycin-A was reductively cleaved to produce 7-deoxyaklavinone and 7-deoxyaklavinone dimer, MA144 E1. Subcellular fractionation indicated that most of the enzyme activity was present in the microsomal fraction and required anaerobic condition and NADPH. The purified enzyme reduced the glycosidic metabolites, MA144 M1 and MA144 N1, as well as aclacinomycin-A. The optimum pH for the anthracycline glycoside reductase reaction using aclacinomycin-A as substrate was 7.4. The enzyme was sigmoidally saturated with aclacinomycin-A and showed the concentration of 1.2 x 10(-4) M required for half maximal activity, and Km value of 7.7 x 10(-5) M for NADPH. The degradative pathway of aclacinomycin-A and its glycosidic metabolites was discussed.


[Effect of plasmid F'Col+trp+ on the transfer of plasmid RP4]. Interaction of conjugative plasmids F'colV colB trp and PR4 in Escherichia coli host was studied during the transfer of the plasmids from cell to cell. The plasmid F'colV colB trp is found to stimulate the transfer of RP4 from the diplasmid strain. This seems to be due to stabilization of the conjugating pairs which require normal pili coded by the plasmid F'colV colB trp.


Activation of proteolytic enzymes during autolysis of disintegrated baker's yeast. Disintegration substantially accelerates autolysis of yeast cells. Three proteases (A, B, and C) take part in the autolytic process, protease A being the activator of the other two enzymes. The role of proteases B and C in the process depends on temperature. At 40 degrees C both proteases are active while at 50 degrees C the major role is played by protease C. At 40 degrees C NaCl acts as inhibitor while at 50 degrees C it activates the process.


Biochemical mechanism of nitrofurantoin resistance in Vibrio el tor. Vibrio el tor cells contain a constitutive reductase enzyme which converts nitrofurantoin to an active principle that is responsible for the observed antibacterial activity of the drug. Acquisition of resistance of this strain towards nitrofurantoin is associated with the loss of this reductase. This enzyme is located in the periplasmic region of the nitrofurantoin-sensitive cells, and seems to play an important role in transporting the drug into the cells.


The forensic identification of semen by isoelectric focusing of seminal acid phosphatase. An isoelectric focusing technique for the separation of acid phosphatases is described. The focusing patterns for semen and vaginal secretion are examined in detail and the technique is evaluated as a means of identifying and distinguishing between these body fluids and other materials of potential forensic interest. The method can reliably be used to diagnose the presence of semen in stains and in the post-coital vagina.


[Therapeutic experiences with clotiazepam (Trecalmo), a new anxiolytic drug from the thienodiazepine group. Results of a field study on 186 patients]. The efficacy and the compatibility of the new anxiolytic Trecalmo (Clotiazepam) were investigated by means of a field trial covering 186 patients, suffering from anxiety and tension as well as from psychosomatic disorders. The new drug proved to be remarkably effective in symptoms as anxiety, restlessness, irritations, tension headache as well as functional cardiovascular diseases and digestive disorders. The compatibility was good.


Studies on the hydrolysis of 1-alkyl-sn-glycero-3-phosphoethanolamine in subcellular fractions of rat brain. The formation of 1-alkylglycerol from 1-alkyl-sn-glycero-3-phosphoethanolamine in different cell fractions of rat brain is reported. The substrates used were labelled either with 14C or 3H in the alkyl residue or with 14C in the alkyl and 3H in the ethanolamine residue. The examination of the lipid- and water-soluble cleavage products showed that both ethanolamine and phosphoethanolamine are liberated from the substrate in the microsomal fraction of 14-day-old rat brain. The latter product is rapidly hydrolyzed. In comparison with other cell fractions, the microsomes contained the highest enzyme activities, which exhibited a pH optimum of 7.1--7.5. SH-group reagents are inhibitors, whereas diisopropylfluorophosphate has no effect. As the animals age, these enzyme activities decrease in brain homogenates.


Ethyl bromoacetimidate, a NH2-specific heterobifunctional reagent. Model reactions with ribonuclease. Ethyl bromoacetimidate was designed as a potential reagent for cross-linking protein NH2 groups with a vicinal nucleophile. The chemical properties of this compound were studied by model reactions with small molecules. Ethyl bromoacetimidate amidinates lysine residues in ribonuclease at pH 9. In a second step, at lower pH values, one of the bromoacetamidino groups bound to the enzyme alkylates a proximal histidine residue. This substitution is pH-dependent with a sharp optimum at 5.6, the same as was earlier observed for alkylation of histidine-119: histidine-12 by halogenoacetates and halogenoacetamides. A common mechanism is suggested for all these types of alkylation. Ethyl bromoacetimidate thus appears as a short-distance crosslinker which can be used, for example, to explore chemically the microenvironment of an essential lysine residue of an enzyme within the active site.


Quantification of non-specific immunosuppressive factors. A simple, reproducible and quantitative method for evaluating certain non-specific immunological inhibitors in a variety of biological fluids is described. Human lymphocytes were stimulated with PHA in the presence of colchicine. Phytohaemagglutinin stimulated a large percentage of cells and colchicine's selective blockage of mitosis limited the stimulated cells to one S phase. These conditions effectively established a maximum amount of DNA synthesis within each culture. Quantification of suppression was then achieved by measuring a decrease from this maximum. The PHA-colchicine assay was successfully used to quantify inhibition by normal plasma, normal mouse sera, mouse neonate sera, murine Ehrlich's and sarcoma I ascitic fluids and an immunoregulatory alpha-globulin peptide preparation. Because of the ability to obtain a specific inhibitory activity for the suppressive factors, this assay was particularly suited for following the isolation of inhibitors during the fractionation of suppressive substances from complex fluids.


Anaerobic isolates in chronic recurrent suppurative otitis media. Treatment with carbenicillin alone and in combination with gentamicin. Tympanocentesis was performed in 32 pediatric patients with chronic recurrent suppurative otitis media. The aspirate was cultured aerobically and anaerobically. Aerobes were isolated from ten patients (31.2%); anaerobes from one patient; and both aerobes and anaerobes from 21 patients (65.6%). There were 46 aerobic isolates. The aerobes commonly recovered were Pseudomonas aeruginosa (24 isolates) Proteus sp. (5) and Staphylococcus aureus (3). There were 32 anaerobes isolated including anaerobic gram-positive cocci (19 isolates) and Bacteroides sp., the latter of which included Bacteroides fragilis group and Bacteroides melaninogenicus (9). The patients were treated by parenteral carbenicillin 300 to 400 mg per kg per day given in four dosages administered for a period of 12 to 21 days (average 17 days). An aminoglycoside (gentamicin) was added in 15 patients. The clinical response was good in 17 patients and poor in 15. There were no side effects or adverse reactions noted during therapy. The above findings demonstrate the polymicrobial etiology of chronic recurrent suppurative otitis media and suggest that treatment directed against the aerobic and anaerobic isolates is efficacious in more than half of the cases.


Hemodynamic characterization of bufuralol-HCl and pindolol based on the competitive effects of isoproterenol. In this hemodynamic study a new beta-receptor blocker, Bufuralol-hydrochloride was compared with Pindolol under an Isoproterenol infusion with increasing doses in healthy male volunteers. We found the following results: 1. Before Isoproterenol peripheral resistance increased after acute i.v. application of Pindolol but decreased after Bufuralol-hydrochloride i.v. application. 2. After beta-receptor blockade with either Bufuralol-hydrochloride or with Pindolol a shift to the right of the dose effect relationship concerning heart rate and cardiac output under Isoproterenol infusion was observed, indicating beta 1-blockade. 3. The reduction of peripheral resistance which is usually observed as a sign of beta 2-blockade was also shifted to the right under the influence of both drugs. 4. This proves Bufuralol-hydrochloride to be a non-specific beta-blocking agent with an affinity to the beta 1- and beta 2-receptors. 5. Although Bufuralol-hydrochloride has a beta 2-blocking property which is even more pronounced than that of Pindolol, it reduces acutely, intravenously given, peripheral resistance.


Lithium-induced nephrogenic diabetes insipidus: studies of tubular function and pathogenesis. We describe a patient with lithium-induced nephrogenic diabetes insipidus in whom detailed investigations of distal tubular function were performed. Clearance of free water during water diuresis was found to be augmented. This suggests proximal suppression of sodium reabsorption by lithium. Reabsorption of free water during high solute clearance was impaired. Acidification of the urine following ammonium chloride loading was abnormal, and this was corrected by sodium sulfate infusion. The cellular mechanism of lithium was investigated by means of indomethacin, an inhibitor of prostaglandin synthesis. Indomethacin caused a partial reversal of the nephrogenic diabetes insipidus, suggesting that the primary cellular action of lithium may be to inhibit the formation of cyclic AMP in the collecting duct cell, although a direct action of indomethacin in increasing solutes in the renal medulla could not be ruled out. It is possible that the lithium-induced polyuria is partially due to an enhancement by lithium of renal prostaglandin action.


A quantitative histochemical technique for the characterisation of alpha-glucosidases in the brush-border membrane of rat jejunum. A quantitative histochemical method to determine the Km and Vmax of alpha-glucosidases in the intestinal epithelium without disruption of the cellular structure is described. 2-Naphthyl-alpha-D-glucoside was used as substrate and hexazonium-p-rosaniline as coupling agent. Using a Leitz MPV2 microdensitometer and a field measuring 4 X 4 micrometers, and reading the test samples against a blank focused on the lamina propria, we observed that the intensity of the colour was a linear function of both the incubation time up to 20 min, and the thickness of the slice up to 20 micrometers. The ratio between the extinction at the absorption maximum and at a second wave-length was constant, whatever the intensity of the colour. By determining the relationship between the extinction and the substrate concentration under standard conditions (slice thickness of of 10 micrometers and incubation time of 10 min), we obtained a saturation curve described by a Km of 0.68 +/- 0.038 mM and a Vmax of 1.41 +/- 0.039 A lambda 480 . 10(-2) . micrometers-1 . min-1. When the hydrolysis of the same substrate by a homogenate of jejunal mucosa was examined biochemically under comparable conditions, a Km of 0.64 +/- 0.012 mM and a Vmax of 57.3 +/- 0.70 mU/mg protein were obtained. When the natural substrate, sucrose, was used in the biochemical study, a Km of 15 +/- 3.5 mM and a Vmax of 149 +/- 24.7 mU/mg protein were obtained. These experiments demonstrate that the kinetic constants of enzyme reactions can be assessed with equal accuracy on histochemical sections as in tissue homogenates.


Intracellular pH regulation of normal and hypertrophic rat myocardium. The myocardial cell pH (pHi) observed during breathing of 0, 7.5, or 10% CO2 in air for 3 h was studied in rats with myocardial hypertrophy due to aortic stenosis and in sham-operated rats. The change in pHi during hypercapnia was significantly smaller in the rats with myocardial hypertrophy, with the apparent nonbicarbonate buffer value (delta [HCO3-]i/delta pHi) being almost three times that of the sham-operated rats. In vitro CO2 equilibrium of myocardial tissue homogenates showed no difference in nonbicarbonate buffer value between homogenates obtained from normal rats and from rats with myocardial hypertrophy. Therefore, it appears that the increased ability of the myocardial cell to regulate its pH during hypertrophy is not due to an increase in the cellular level of nonbicarbonate buffers, but seems to be related to a larger bicarbonate uptake by the myocardial cell during hypercapnia.


Effect of cerebral extracellular fluid acidity on total and regional cerebral blood flow. Total and regional cerebral blood flow (CBF), and cerebrospinal fluid (CSF), and arterial blood acid-base status were measured in 26 chloralose-urethan-anesthetized dogs before and after 30 and 60 min of ventriculocisternal perfusion with artificial CSF equilibrated with 7% CO2 and containing either low (8.7 or 9.1 meq/l), normal (19.6 meq/l), or high (34.7 meq/l) bicarbonate ion concentration ([HCO3-]). An inverse linear relationship was observed between the CSF pH and total CBF. Regional blood flow changes were greater in structures that were closest to the ventricular system. In addition, regional blood flow changes were greater in all tissues studied after 60 min of perfusion than after 30. Perfusion with the control [HCO3-] caused no significant changes in either acid-base status or CBF. We believe that the regional cerebral blood flow changes are the result of changes in the H+ concentration gradient across the cerebral extracellular fluid (ECF) space due to the diffusional exchange of HCO3- between CSF and ECF. It is concluded that cerebral ECF acidity is important in the local regulation of cerebral blood flow.


Aortic body chemoreceptor responses to changes in PCO2 and PO2 in the cat. Responses of aortic chemoreceptor afferents to a range of arterial carbon dioxide tension (Paco2) changes at various levels of arterial oxygen tension (Pao2) were investigated in 18 cats anesthetized with alpha-chloralose and maintained at 38 degrees C. Aortic chemoreceptor activity, end-tidal oxygen pressure, end-tidal carbon dioxide pressure, and arterial blood pressure were continuously monitored. Arterial blood gases were measured in steady states. Single or a few clearly identifiable afferents were studied during changes and steady states of Pao2 and Paco2. All the aortic chemoreceptor afferent discharge rates increased with Paco2 increases from hypercapnia (10-15 Torr) to normocapnia and moderate hypercapnia (30-50 Torr) and with Pao2 decreases from above 400 to 30 Torr. Hypoxia augmented the response to Paco2 most effectively in the range of 10-40 Torr. At any Pao2, the discharge rate reached a plateau with sufficient intensity of hypercapnia. The Paco2 stimulus threshold at a Pao2 of 440 Torr was about 15 Torr, and at a Pao2 of 60 Torr it was 10 Torr. In the transition from hypocapnia to hypercapnia, responses increased gradually, usually without an overshoot. The steady-state responses to Paco2 of the majority of aortic chemoreceptors resembled those of carotid chemoreceptors. The responses of both receptors can be attributed to the same basic type of mechanism.


Effect of increased blood-oxygen affinity on oxygen transport in hemorrhagic shock. Effect of increased blood-oxygen affinity on tolerance of hemorrhagic shock was studied in pentobarbital-anesthetized rats. Rats were first exchanged transfused with blood whose P50 had been reduced by various methods by 4-21 Torr. Hypotension (BP = 30 Torr) was induced and maintained at this level by controlled hemorrhage; it was terminated when reinfusion of shed blood became necessary to sustain this blood pressure. Initial rate of bleeding during shock was inversely proportional to P50, varying from 0.52 ml.min-1.kg-1 in controls to 1.5 ml.min-1.kg-1 in the group with the lowest P50, a reaction probably indicating increased sympathetic output in the latter group. Duration of shock tolerance varied from 50 +/- 16 min in controls to 28 +/- 11 min (SD, P less than 0.001) in the group with the lowest P50. Central venous SO2 (SCVO2) and PO2 (PCVO2) were significantly higher and lower, respectively, in low-P50 animals than in controls, probably because of limited oxygen extraction due to increased blood oxygen affinity. VO2 and cardiac output were significantly lower, and mortality was significantly greater, in low-P50 animals. The data suggest that a left shift of the oxygen dissociation curve limits oxygen delivery during hemorrhagic shock.


Postcapillary pH disequilibrium after gas exchange in isolated perfused liver. pH equilibration after gas exchange in a systemic vascular bed was investigated using an isolated guinea pig liver preparation perfused with a blood-free Krebs-Ringer-bicarbonate solution. Effluent perfusate was withdrawn into a stopped-flow apparatus in which pH and temperature were continuously monitored. A decrease in pH of the perfusate after exit from the liver was observed. Addition of acetazolamide to inflowing perfusate had little effect on this fall in effluent fluid pH, while the addition of carbonic anhydrase completely abolished the decrease in pH. These results suggest that: 1) after the addition of metabolically produced CO2, the reaction CO2 leads to H2CO3 leads to H+ + HCO3- in the perfusate did not reach equilibrium during passage through the liver; 2) acetazolamide had little effect on the magnitude of the pH disequilibrium; and 3) carbonic anhydrase accelerated the reaction to equilibrium. We conclude that little or no catalysis of the conversion of perfusate CO2 to H2CO3 takes place within the hepatic circulation and that the presence of slow postcapillary blood pH changes in vivo may be dependent on the specific species and organ vascular bed from which the blood exited after participating in gas exchange.


Tracheal fluid in fetal lambs: spontaneous decrease prior to birth. We studied tracheal fluid (TF) production in 14 fetal lambs: 6 controls, 6 receiving atropine on 1 or more of the last 7 days before birth, and 2 with bilateral section of the cervical vagosympathetic trunk. A cannula diverted all TF into an intrauterine bag; we collected TF intermittently and measured its volume. All ewes delivered spontaneously at 128-150 days' gestation. TF production decreased before birth in all fetuses except one control. TF production decreased before birth in all fetuses except one control. TF production did not correlate with fetal arterial blood gas tensions, hematocrit, or plasma proteins. In controls only, TF production correlated with fetal arterial pH (P less than 0.02); however, the pH range was small and the correlation has questionable physiological significance. For all fetuses, TF production during the 7 days before birth correlated inversely with the plasma cortisol concentration of 48 h previously (n = 36; r = -0.603; P less than 0.001). We conclude a) TF production in fetal lambs decreases before spontaneous term or preterm labor; b) this decrease is not affected by atropine or by section of the cervical vagosympathetic trunk; and c) the decrease in TF production may be related to increased secretion of cortisol.


Presteady state kinetics of trypsin-catalyzed hydrolyses of dansyl-arginine derivatives. Interactions between trypsin and each of five dansyl-arginine derivatives, dansyl-L-arginine methyl ester (L-DAME), dansyl-D-arginine methyl ester (D-DAME), dansyl-L-arginine amide (L-DAA), dansyl-L-arginine (L-DA), and dansyl-D-arginine (D-DA), are accompanied by a fluorescence intensity change which can be followed by the stopped-flow method. These compounds are substrates or products in trypsin-catalyzed hydrolysis reactions. All of these compounds, except L-DAA, show a considerable fluorescence intensity increase in the reaction with trypsin. The observed rate constant, tau obsd -1, for the initial fluorescence intensity enhancement in the reaction between trypsin and D-DAME yields a typical hyperbolic curve when the rate is plotted as a function of the ligand concentration. This result is consistent with a two-step mechanism (1) in which a fast bimolecular association process is followed by a slower unimolecular isomerization process. The isomerization process may be considered to be associated with a conformational change of the enzyme molecule, induced by the formation of the enzyme-substrate complex (1). The rate of the isomerization process depends on pH. The rates obtained for L-DAME and D-DAME increase linearly with decrease of the hydrogen ion concentration in the pH range below neutral.


Purification and characterization of an endonuclease specific for single-stranded DNA from Bacillus subtilis Marburg. Bacillus subtilis Marburg TI (thy,trpC2) has at least four endonuclease activities as assayed by measuring the conversion of single-stranded circular f1 DNA to the linear form by agarose gel electrophoresis. One of them, which is specific for single-stranded DNA (named endonuclease MII), was purified about 320 times by two chromatographic steps and gel filtration, thereby eliminating exonuclease and phosphomonoesterase activities. This activity requires divalent cations but does not require ATP. The molecular weight estimated by gel filtration was about 57,000 daltons. The cleavage products have 5'-phosphoryl termini. At low concentrations, double-stranded DNA is not split to any detectable extent. At high concentrations, however, double-stranded superhelical DNA is attacked to yield open-circular and linear DNA's. The activity of the enzyme towards single-stranded circular DNA relative to that towards double-stranded linear DNA was calculated to be approximately 5,000:1 by comparing the initial rates of introducing single-strand breaks into the DNA's.


An esterase on the outer membrane of Pseudomonas aeruginosa for the hydrolysis of long chain acyl esters. A new esterase activity which hydrolyzes palmitoyl-CoA was found in the membrane fraction of Pseudomonas aeruginosa. All the 11 strains of P. aeruginosa tested possessed this esterase activity. The esterase was constitutive and was fully active on the intact cell bodies toward substrates in the medium. It was located on the outer membrane of the cell envelope, and was not released into the culture medium. This activity was designated as OM (outer membrane) esterase. OM esterase was solubilized from the cell envelope with EDTA-Triton X-100 and purified 690-fold. It was a minor component of the outer membrane. Its molecular weight was approximately 55,000. The activity was rather stable to heat, a wide range of pH, and treatment with detergents and organic solvents. No cofactors were required. The pH optimum of the reaction was 8.5. Among various acyl-CoAs, only long chain (C12--C18) thioesters were hydrolyzed. OM esterase also hydrolyzed some kinds of oxy-esters such as p-nitrophenyl acyl esters, monoacyl esters of sucrose and Tween 80 (polyoxyethylene sorbitan monooleate). On the other hand, triglycerides, phospholipids, or hydrophobic monoesters were not hydrolyzed at all. Thus, this enzyme seems to have specificity for long chain acyl esters with hydrophilic groups, whether thio- or oxy-ester. Mutants deficient in this esterase activity were isolated. These mutants were unable to grow on Tween 80 as a sole carbon source. This suggests a possible role of OM esterase in the utilization of acyl esters as carbon sources.


Occurrence of old yellow enzyme in Gluconobacter suboxydans, and the cyclic regeneration of NADP. Old yellow enzyme system has been found in the cytosol fraction of Gluconobacter suboxydans. This is the first time that the enzyme has been found in organisms other than yeast cells. Old yellow enzyme [EC 1.6.99.1], D-glucose-6-phosphate dehydrogenase [EC 1.1.1.49], and catalase were isolated and crystallized separately from the organism. The old yellow enzyme from G. suboxydans showed catalytic and physicochemical properties almost identical with those of the enzyme from yeast cells. NADPH was specifically oxidized by the old yellow enzyme and the reduced enzyme was spontaneously reoxidized by atmospheric oxygen. The old yellow enzyme from G. suboxydans also contained FMN as a prosthetic group, and two mol of FMN were found per mol of enzyme (molecular weight, 88,000 as determined by gel filtration). In the oxidation of D-glucose-6-phosphate to 6-phospho-D-gluconate, cyclic regeneration of NADP occurred smoothly in the presence of D-glucose-6-phosphate dehydrogenase and catalase, even when a limited amount of NADP or NADPH was present in the reaction mixture.


Calcium binding of troponin C. I. A potentiometric titration study. Structural changes of troponin C on calcium binding were studied by hydrogen ion titration, circular dichroism, and fluorescence measurements. The potentiometric titration curves in the carboxyl region are shifted towards lower pH with calcium binding. The intrinsic pK of the carboxyl groups at the calcium binding sites decreases by 0.8 pK unit on calcium binding; on the other hand, magnesium ions have little effect on the intrinsic pK of the carboxyl groups. The intrinsic pK of the imidazole group is not affected by calcium binding. The value of w, an electrostatic interaction factor, is identical for calcium-free and calcium-bound troponin C and is about half of the value calculated assuming a compact sphere. The results of difference titration on the calcium binding indicate that the pH of troponin C solution increases on addition of CaCl2 up to 2 mol of Ca2+ per mol of troponin C and then decreases on further addition of CaCl2. The pH increase is depressed in the presence of MgCl2, in the low pH region, or at high ionic strength. The pH increase is also observed on addition of MgCl2. The ellipticity at 222 nm was measured under the same conditions as the difference titration measurements, and the relation between the pH change and the conformational change of troponin C on calcium binding is discussed based on the results obtained. The number of calcium binding sites and the binding constants estimated by analysis of these difference titration curves were in agreement with the results of Potter and Gergely (22). No magnesium binding site was observed. The tyrosine fluorescence measurements indicated that the binding site near tyrosine-109 is one of the high affinity sites.


Multiple forms of ribonuclease H from rat liver cytosol. Three forms (termed I, II, and III) of ribonuclease H (RNase H) [EC 3.1.4.34] activity are present in rat liver cytosol. These enzymes degrade RNA specifically in RNA-DNA hybrid structures. They were eluted at 0 M, 0.25 M, and 0.5 M KCl in phosphocellulose chromatography, and were further purified by using blue Sepharose. They are further distinguished from one another by their ionic requirements, optimal pH, molecular weights, sedimentation coefficients, and sensitivity to the -SH reagent, p-chloromercuribenzoate, although I and III have similar characteristics. They liberate a mixture of oligonucleotides with 5'-phosphate and 3'-hydroxyl termini.


Adenosine triphosphate-dependent calcium uptake of synaptic vesicle fraction is largely due to contaminating microsomes. Ca2+ uptake by synaptic vesicle fractions isolated from bovine caudatolenticular nuclei and from rat brain was studied. The purified vesicle fractions from both materials took up very little Ca2+ even in the presence of ATP and Mg2+, but the crude fractions took up Ca2+ actively, showing the maximum uptake around pH 7.0. Since the crude fractions were contaminated by microsomes, which are known to accumulate Ca2+ actively (Yoshida, H., Kadota, K., & Fujisawa, H. (1966) Nature 212, 291--292; Otsuka, M., Ohtsuki, I., & Ebashi, S. (1965) J. Biochem. 58, 188-190), the active uptake of Ca2+ appeared to be largely, if not wholly, due to microsomal contamination.


Changes in the random distribution of sialic acid at the surface of the myeloid sinusoidal endothelium resulting from the presence of diaphragmed fenestrae. Diaphragmed fenestrae (DF) are sites of increased vascular permeability. The anionic charge distribution at the luminal aspect of the DF of the endothelium of the bone marrow vessels has been studied after aldehyde fixation by means of colloidal iron (CI), native ferritin (NF), and polycationic ferritin (PCF). At pH 1.8, these cationic agents are bound by the nonmodified luminal endothelial cell surface but not at the sites of the DF. PCF was used over a pH range of 1.8--7.2 (CI is unstable at higher pH levels, whereas NF which has a pI of 4.5 is anionic above this point). PCF shows increased binding at the DF from pH 3.5 upwards. PCF binding at pH 1.8 at the nonmodified luminal cell surface is significantly diminished by neuraminidase treatment which, however, does not perceptibly reduce PCF binding at the higher pH levels. It is concluded that there are exposed sialic acid groups at the lunimal cell surface which are absent or significantly fewer at the sites of the DF, whereas other anionic materials possibly with a pKa higher than that of sialic acid (pKa 2.6) are present both at the DF and at the nonmodified endothelial cell surface.


pH regulates the polymerization of actin in the sea urchin egg cortex. The state of actin in the isolated cortex of the unfertilized sea urchin egg can be controlled by experimentally manipulating the pH of the isolation medium. Cortices isolated at the pH of the unfertilized egg (6.5--6.7) do not contain filamentous actin, while those isolated at the pH of the fertilized egg (7.3--7.5) develop large numbers of microvilli which contain bundles of actin filaments. Cortices that are isolated at pH 6.5 and then transferred to isolation medium buffered at pH 7.5 also develop actin filaments. However, the filaments are not arranged in bundles and microvilli do not form. Although the cortical granules in cortices isolated at pH 6.5 discharge at a free Ca++ concentration of approximately 10 micrometer, actin polymerization is not induced by increasing the Ca++ concentration of the isolation medium. These results suggest that the increase in cytoplasmic pH which occurs following fertilization induces the polymerization of actin in the egg cortex.


Biochemical and coagulation changes after clamping and declamping of the superior mesenteric artery in dogs. Intestinal ischemia produced by S.M.A. temporary occlusion, results in a degree of metabolic and coagulation changes at the ischemic area. When the blood flow is restored, those changes become manifest in the systemic circulation. Both the degree of those changes and their effects are documented and discussed.


A growth factor from spinal cord.

A growth factor from spinal cord. Mitogenic activity is present at a variety of sites in the central nervous system. A growth factor was purified from neonatal bovine spinal cord. It has a pI of 9.5-9.8 and a molecular weight of about 11,000 daltons. Spinal cord growth factor is a basic polypeptide that is inactivated by extremely acid or basic conditions. Its mobility on SDS polyacrylamide gels suggests that this factor is different from pituitary FGF and brain FGF-1.


Plasma beta-endorphin and beta-lipotropin in the human fetus at delivery: correlation with arterial pH and pO2. Beta-endorphin-like immunoactivity was measured in the umbilical cord plasma of 45 term human fetuses. Mean concentration was 91 +/- 16 (SEM) pg/ml,an the normal adult level of 30.7 +/- 2.7 pg/ml. This immunoactivity was further characterized in 10 cases by Sephadex G-50 chromatography to separate beta-endorphin from beta-lipotropin (beta-LPH). Mean beta-endorphin and beta-LPH concentrations were 57 +/- 12.8 and 455 +/- 101 pg/ml, respectively. Both were higher (P less than 0.01) than the mean beta-endorphin and beta-LPH concentrations reported in the adult. The mean molar beta-endorphin to beta-LPH ratio was 0.35 in the fetus and 0.36 in the adult. In 17 fetuses whose umbilical arterial and venous concentrations were measured separately, mean beta-endorphin-like immunoactivity was higher in the artery than in the vein. A highly significant negative correlation (r = -0.831; P less than 0.001) was present between umbilical arteiral pH and beta-endorphin-like immunoactivity. A negative correlation (r = -0.611; P less than 0.005) with arterial pO2 was also noted. We conclude that high levels of beta-endorphin-like immunoactivity, composed of both beta-endorphin and beta-LPH, circulate in the human fetus at term, and that hypoxia and secondary acidosis may be major stimuli to the release of these peptides.


Fluorescent antibody test kit for rapid detection and identification of members of the Bacteroides fragilis and Bacteroides melaninogenicus groups in clinical specimens. The Fluoretec fluorescent antibody test kit (Pfizer Inc., New York, N.Y.), developed for the rapid detection of members of the Bacteroides fragilis and B. melaninogenicus groups, was evaluated by testing 58 stock cultures and 76 clinical specimens. The test reagents detected 100% of 40 B. fragilis and B. thetaiotaomicron stock culture strains, although only 22% of 18 B. vulgatus, B. distasonis, and B. ovatus strains showed positive fluorescence. The 76 clinical specimens were evaluated by examining fluorescent antibody-stained smears of 49 specimens of purulent material and smears of 27 blood cultures which were positive for gram-negative bacilli by Gram stain or subculture. The fluoretec reagent detected members of the B. fragilis group in 28 (97%) of the 29 specimens of purulent material and all (100%) of the 16 blood cultures in which these anaerobes were demonstrated by culture. Overall, the Fluoretec reagent detected members of B. fragilis group in 44 (98%) of the 45 clinical specimens which were shown by culture to contain these anaerobes. Two of the 76 clinical specimens gave positive fluorescence for members of the B. fragilis group but failed to yield these organisms by culture. Members of the B. melaninogenicus group were detected by culture in 15 specimens and in each case their presence was demonstrated by the Fluoretec reagent. No members of the B. melaninogenicus group were isolated from five clinical specimens that gave positive fluorescence with the B. melaninogenicus reagent.


Effects of cell culture and laboratory conditions on type 2 dengue virus infectivity. The stability of type 2 dengue virus to exposure to a variety of laboratory conditions was determined. Suckling mouse brain passage virus was adapted for growth in BHK-21 cells, and plaque assays were performed using a tragacanth gum overlay. A three- to fourfold increase in plaque size could be obtained if monolayers were subconfluent at time of inoculation. Incubation of virus for 24 h at 37 degrees C, pH 6.5, or in buffer containing 1 mM ethylenediaminetetraacetate considerably reduced virus infectivity as compared with virus incubated for the same period at 4 degrees C, pH 8.0, or in buffer with or without 1 mM CaCl2 and 1 mM MgCl2. Multiple freezing and thawing of virus tissue culture medium containing 10% fetal calf serum did not reduce virus infectivity.


Relaxation therapy, desensitization, and the treatment of anxiety-based disorders. Evaluated systematic desensitization and relaxation training for the treatment of snake phobia and test anxiety as representatives of two classes of anxiety-based disorders. Treatment outcomes were assessed by examining situational and dispositional components of anxiety as related to these disorders and by behavioral measures of performance in relevant anxiety-provoking situations. Analyses of variance revealed that more pervasive anxiety reductions occurred for the more focalized animal phobia and that there was little difference in the effectiveness of desensitization and relaxation training. The generalizability of research findings based on the treatment of animal phobias was questioned, and the possible role of nonspecific factors in determining success was considered.


Rapid diagnosis of anaerobic infections by gas-liquid chromatography. It was postulated that the short chain fatty acids (SCFA) produced by anaerobic bacteria might serve as microbial markers in purulent material. Eighteen pus specimens from various sources were analysed by gas-liquid chromatography (GLC), and the SCFA detected were compared with the microorganisms isolated by conventional methods. It was found that the detection of propionic, isobutyric, butyric, or isovaleric acids by direct GLC of pus specimens is strong evidence for anaerobic infection but not specific for Bacteroides fragilis. It was also shown that the presence of succinic acid in pus specimens does not necessarily indicate infection by anaerobes. It can be concluded that direct GLC of purulent material provides a rapid and reliable presumptive method for the differentiation between anaerobic and aerobic infections.


Serum gamma-glutamyltransferase isoenzymes in extrahepatic biliary obstruction. The gamma-glutamyltransferase isoenzymes in the sera of patients with extrahepatic biliary obstruction have been studied, using electrophoretic, gel filtration, and ultracentrifugation techniques, and compared with those present in normal sera. Five isoenzymes were shown to exist in patients' sera, three of which were not demonstrated in normal sera. The observations are discussed in relation to the influence of biliary regurgitation and the possible solubilisation of membrane-bound enzymes. The results are compared with those of previous studies on alkaline phosphatase.


Long-term use of a "stable" bicarbonate containing dialysate. We present an original method for the preparation of "stable" dialysate containing 35 mEq/l of bicarbonate. The dialysate was utilized with 4 patients for periods ranging from 4 months to 1 year according to a short-term recirculated dialysis schedule in closed circuit (20-40L) (2-2 1/2 hrs) on alternate days. Preliminary results are reported here with respect to the tollerance of the dialytic run and correction of the acid-base balance equilibrium. The clinical tollerance is excellent despite high dehydration rates even in patients particularly sensitive to ultrafiltration. The acidosis correction would seem to be much better with bicarbonate than with traditional dialysis. The difference is even higher if we consider the brevity of the dialysis. During the bicarbonate dialysis we do not observe any fall of the PCO2 or significant difference in PO2 in the patient's blood. The correction of acidosis probably causes the normalization of pre-dialytic potassiemia in spite the "net" removal of K with short dialysis is considerably less.


Effect of acetate upon arterial gases.

Effect of acetate upon arterial gases. Following infusion of sodium acetate, a significant fall in arterial oxygen tension (PO2) was observed (p less than 0.001). A rise in PCO2 or bicarbonate was also statistically significant. pH was however remained unchanged. Though a rise in PCO2, bicarbonate, pH was observed following infusion of sodium bicarbonate, no significant fall in PO2 was observed. These results suggest a role of acetate ion for the fall of arterial oxygen tension; the hemodialysis-induced hypoxemia.


Dialyzing room disinfection with ultra-violet irradiation. Infections represent a major problem in dialysis treatment, thus the dialyzing room should be kept abacterial as possible. We have installed 15-watt ultra-violet (U-V.) lamps for every 13.5 m2 on the ceiling for the purpose of the room disinfection and used them for 16 hours nightly after working hours. Bacteria were killed with over 10 hours irradiation even at the areas of low U-V. intensity where the irradiation may not be direct. This unexpected effectiveness might be from the influence of reflected rays and 03 produced. When half the lamps were turned on, the bacteriocidal effect was not sufficient in some areas. Any living organism with nucleic acids must be inactivated by this treatment, for the baceteriocidal effect is due to the nucleic acids injury. Furthermore, safety, readiness after the treatment, easy application and the negligible costs would make this method more advantageous to the other methods in room disinfection.


Clinical and microbiological aspects of chemotherapeutic agents used according to the specific plaque hypothesis. Certain forms of dental decay and periodontal disease appear to be due to specific bacterial infections following overgrowth of certain indigenous plaque bacteria, i.e., the specific plaque hypothesis, If so, then antimicrobial treatment based on a diagnosis of elevated levels or proportions of these organisms should be considered. Such treatment cannot be administered according to concepts of the non-specific plaque hypothesis. A treatment philosophy is presented which is based upon considerations long established in medical infections following overgrowth of certain indigenous plaque bacteria, i.e., the specific plaque hypomicrobial to the site of the infection for periods long enough to suppress or destroy the pathogenic agent. Examples of this treatment philosophy are given.


Summary of an International Symposium on phenytoin-induced teratology and gingival pathology. Phenytoin has been the preferred drug for treating grand mal epilepsy for more than 40 years. The metabolism of this drug, its teratogenic potential, and its role in the pathogenesis of gingival overgrowth are discussed.


Biologic correlates of psychoanalytic concepts. In deference to Freud's ingenious predictions of the future potential of the biological sciences concerning psychoanalytic tenets and clinical disorders, an attempt is made to bring the concepts of the Three Polarities, life and death instincts, the Unconscious, and repression into association with relevant anatomical, neurophysiological, and neurochemical data. Implications for diagnosis and therapy were considered in the light of recent and anticipated neurobiological advances. Building bridges between biologic and physiologic processes is exceedingly complicated by the enormous complexities of both fields. There is good reason to hope that the speculative nature which at this time pervades our bridging efforts will eventually be substituted by unequivocal facts and deductions.


The fate of acephate and carbaryl in water. Acephate was resistent to hydrolysis in distilled, buffered water at pH 4.0 to 6.9, but not at pH 8.2, held for 20 days at 20 or 30 degrees C. The maximum conversion to methamidophos was 4.5% of the added acephate at pH 8.2 and 20 degrees C. The persistence of acephate in two natural waters, held at 9 degrees C for up to 42 and 50 days varied: 80% were recovered from pond water after 42 days, and 45% from creek water after 50 days. Rates of acephate degradation increased greatly when treated water samples were incubated in the presence of sediments, but not if water and sediment were autoclaved prior to treatment and incubation. The greatest conversion to methamidophos, 1.3% of the added acephate, had occurred after 42 days in pond water without sediment. Under the same conditions, carbaryl was less persistent than acephate in the natural waters: 18 to 20% were recovered from pond water after 42 days, and 37 to 40% from creek water after 50 days. The presence of sediment did not affect its degradation significantly. But more than 55% were recovered after 50 days if water and sediment were autoclaved prior to treatment and incubation. Neither acephate, methamidophos, nor carbaryl could be shown to escape from water into the atmosphere.


Transmitter mediated arginine vasopressin release from superfused hypothalamus and pituitary gland. The study was designed to investigate the effect of various neurotransmitters on the hypothalamus and pituitary gland to determine the sites of their action. Superfused isolated rat hypothalami and pituitary glands demonstrated basal secretion of arginine vasopressin (AVP) and repeated response to stimulation thus showing the viability of the preparation. Acetylcholine and histamine stimulated the release of AVP at the hypothalamic and pituitary levels; dopamine and norepinephrine released AVP in a dose related manner only from the hypothalamus; angiotensin II released AVP in the same fashion only from the pituitary gland. AVP secretion stimulated by dopamine and norepinephrine may represent synaptic inputs which are localized at the hypothalamus and must be distinguished from the site of action at the pituitary gland of angiotensin II.


Chemical modification of horseradish peroxidase. Preparation and characterization of tracer enzymes with different isoelectric points. Horseradish peroxidase (HRP), a plant glycoprotein with a molecular weight of 40,000 D and a molecular radius (ae) of 30 A, has been modified chemically to prepare tracer molecules with different molecular charge. Modification of free carboxyl groups on the enzyme is achieved by carbodiimide activation and subsequent reaction of activated carboxyl groups with a nucleophile; uncharged groups or radicals containing additional positively charged moieties are introduced into the protein molecule resulting in an increased net positive charge of the tracer. Amino groups in the protein molecule are modified by acetylation or succinylation; this reaction will increase the net negative charge of the enzyme by either introducing an uncharged group or an additional carboxyl radical. The tracer molecules so obtained are then characterized in terms of molecular size and charge by column chromatography and isoelectric focusing respectively. The enzymatic activity as measured by 3,3'-diaminobenzidine reaction, the pH optimum and the absorption spectra for the modified enzymes remain virtually unchanged.


Elevated proteinase activities in mouse lung tumors quantitated by synthetic fluorogenic substrates. Proteinase activities in malignant and normal lung tissues were measured using two synthetic substrates that consist of a fluorophor coupled to a peptide moiety. The hydrolysis of CBZ-Val-Lys-Lys-Arg-4-methoxy-2-naphthylamide and BZ-Gly-Gly-Arg-4-methoxy-2-naphthylamide were studied in homogenates of two types of mouse lung tumors, the Lewis lung tumor of the C57 black mouse and the KHT tumor of the C3H mouse. The activity of CBZ-Val-Lys-Lys-Arg-4-methoxy-2-naphthylamide hydrolysis had a pH optimum of 6.3 and a Km of 2.1 x 10(-4) M, required a thiol activator, and was inhibited by leupeptin suggesting the activity of a cathepsin B-like enzyme. The activity of BZ-Gly-Gly-Arg-4-methoxy-2-naphthylamide hydrolysis had a pH optimum of 6.7 and a Km of 3 x 10(-5) M. Lung tumor homogenates contained higher hydrolytic activities for both substrates than normal lung homogenates.


Discontinuous density gradient separation of human mononuclear leucocytes using Percoll as gradient medium. The use of a new commerically available medium (Percoll) for fractionation of human mononuclear leucocytes is described. Cells can be fractionated on the basis of their densities with high reproducibility. The separated cells were characterized by morphological and functional criteria. Monocytes can be obtained in the low density fractions with a purity of 70%--90%. Lymphocytes were found in high density fractions with a purity up to 99%. No separation between E-rosette forming (E-RFC) and surface immunoglobulin-bearing lymphocytes was obtained. However, a reduced number of high avidity E-rosette forming lymphocytes (HAE-RFC) was found within low density lymphocytes. Best spontaneous DNA synthesis and reaction in mixed leucocyte culture (MLC) were obtained with cells isolated from the 1.066/1.068 density interface, whereas the stimulation with phytohemagglutinin (PHA) or pokeweed mitogen (PWM) had a peak response with cells from the 1.064/1.066 density interface. Colony forming myelopoietic stem cells and colony forming T-lymphocytes were detected in fractions of low density.


Hepatic enzymes of tyrosine metabolism in tyrosinemia II. A middle-aged adult male with a mild form of tyrosinemia II (Richner-Hanhart syndrome) is described. Treatment with a low-tyrosine diet caused a fall in plasma tyrosine and clearing of the hyperkeratosis of the soles. Liver biopsy of this patient revealed low but measurable levels of cytoplasmic tyrosine aminotransferase and elevated levels of the mitochondrial tyrosine-metabolizing enzyme aspartate aminotransferase. It is hypothesized that these enzymes have been induced in sufficient amounts to account for the mild clinical course.


Metabolic control of neuronal pacemaker activity and the rhythmic organization of central nervous functions. The endogenous rhythmic activity of isolated pacemaker neurones of Aplysia californica appears to be controlled by the operation of a substrate cycle. The recycling of fructose-6-phosphate is mediated by two membrane-bound enzymes: phosphofructokinase (PFK) and fructose-1,6-diphosphatase (FDPase). Allosteric effectors which promote the PFK-FDPase system either increase the regular beating activity or induce bursting discharges, while inhibitory effectors reduce pacemaker activity. Associated with the PFK-FDPase cycle are slow oscillations in membrane potential, the postulate being that changes in amplitude and time period of the waves are brought about by the cyclic fluctuations of H+ ions and ATP in the immediate vicinity of the membrane. Other enzyme reactions which affect the concentrations of gluconeogenic substrates or PFK effectors can modulate the oscillatory driving input, a good example being the neurogenic amino acid glutamate. Modifiers of FDPase and PFK are equally effective in changing pacemaker activity within the intact neuronal network and, hence, the rhythmic body function connected to this network. This has been demonstrated with pacemaker neurones governing cardiovascular activity in Apylsia, blood pressure or heart beat in the cat, and respiration or thermoregulation in the rabbit. Nature appears to have achieved a functional differentiation between different pacemaker neurones by altering their response to at least one or two of the PFK and FDPase effectors. New periodicities can be entrained by current stimuli on the pre-existing rhythms of isolated Aplysia pacemaker neurones. Stimulus-induced resetting of the discharges is in fact accompanied by a redistribution between two kinetically distinct forms of PRK, and modifiers of this enzyme can stabilize the new periodicities or facilitate the conditioning effect of a stimulus. Memory facilitation and consolidation under PFK modifiers could also be demonstrated in avoidance and discrimination learning trials with honey bees and rats, which are consistent with the metabolic nature of the slow-wave rhythmicity in vertebrate microneurones thought to be the site of memory storage.


Oscillations of cyclic nucleotide concentrations in relation to the excitability of Dictyostelium cells. Aggregating cells of Dictyostelium discoideum are able to release cyclic AMP periodically. The oscillations of cAMP generation are associated with changes in adenylate cyclase activity. Cyclic AMP receptors on the cell surface are functionally coupled to the oscillating system as evidenced by phase shifts that are induced by small pulses of extracellular cAMP. An important element of the oscillating system is the signal processing from surface receptors to the adenylate cyclase. This pathway exhibits adaptation resulting in the suppression of responses to constant, elevated concentrations of cAMP. The signal input for adenylate cyclase activation is, therefore, a change in the extracellular cAMP concentration with time. Oscillations in the absence of detectable changes of intra- or extracellular cAMP concentrations suggest the possibility that there is a metabolic network in D. discoideum cells that undergoes oscillations without coupling to adenylate cyclase. Cyclic GMP concentrations oscillate with a slight phase difference in advance of that of cAMP, suggesting that the two nucleotide cyclases might not be activated by the same mechanism. Elevation of extracellular calcium exerts an inhibitory effect on the accumulation of cAMP and on the second of the two cGMP peaks.


Membrane potential oscillations in molluscan "burster" neurones. Membrane potential oscillations can be induced in molluscan neurones under a variety of artificial conditions. In the so-called 'burster' neurones oscillations are generated even in isolated cells. A likely mechanism for 'bursting' involves the following ionic currents: 1. A transient inward current carried by Na+ and Ca2+. This current is responsible for the upstroke of the action potentials. 2. A delayed outward current carried by K+. This current is voltage-sensitive and is responsible for the downstroke of the action potential during the early part of the burst. It becomes progressively inactivated during the burst. Its amplitude depends on the intracellular pH. 3. A rapidly developing outward current carried by K+ which is inactivated at potentials close to action potential threshold. This current tends to hold the membrane in the hyperpolarized state and is involved in spacing the action potentials. 4. A prolonged inward current which may not inactivate. It is probably carried by both Na+ and Ca2+. This current is responsible for the depolarizing phase of the burst but also contributes to the action potential. 5. A slowly developing outward current, carried by K+. This current appears as a result of a slow increase in intracellular ionized calcium and is responsible for the hyperpolarizing phase of the burst. Note that a transient increase in this current may also contribute to the falling phase of the action potential during the later stages of the burst. It is also sensitive to intracellular pH. One of the more significant features of this system of producing membrane potential oscillations is that the frequency of the bursts depends on the rate at which the intracellular ionized calcium returns to its resting level. This process depends on the metabolic state of the animal which can thereby exert a considerable influence on the electrical activity of burster neurones.


Specificity of serotoninergic inhibition in Limulus lateral eye. The receptor specificity for synaptically mediated lateral inhibition in Limulus lateral eye retina was studied by structure-activity correlations of the action of the putative indoleaminergic neurotransmitter, serotonin (5-HT), and its isomers and structural analogs, tryptamine (TRYP), 6-hydroxytryptamine (6HT), 5,6-dihydroxytryptamine (5,6-DHT), 5-hydroxydimethyltryptamine (5-HDMT), and 5-hydroxytryptophan (5-HTP). The 5-HT blockers, lysergic acid diethylamide (LSD), bromo-LSD (BOL), and cinanserin, were also tested. The inhibitory action of the indoleaminergic agonists is highly structure-specific. An hydroxyl group in the 5 position of the indole nucleus, sterically unencumbered by hydroxyls in neighboing positions, is essential. In order of decreasing potency, 5-HT, 5-HDMT, and 5-HTP are active agonists; TRYP, 6-HT, and 5,6-DHT are inactive. Configuration and mobility of the side chains of the active agonists also affect the interaction, and these side-chain characteristics correlate with agonist potency. The receptors for inhibitory action and for transmembranal transport in reuptake are different. Both active agonists and inactive analogs appear to be taken up (Adolph and Ehinger, 1975. Cell Tissue Res. 163:1-14). LSD and BOL have bimodal actions: direct inhibition and agonist blockade. These actions may be mediated via low-specificity presynaptic uptake receptor sites rather than highly specific, postsynaptic, agonist receptor sites.


The treatment of psychotic major depressive disorder with drugs and electroconvulsive therapy. A retrospective chart review of 54 patients demonstrating depression with psychotic symptoms was accomplished with the use of Research Diagnostic Criteria (RDC) for diagnosis of psychotic major affective disorder. Patients received adequate trials of either tricyclic antidepressants alone, antipsychotics, the two in combination, or electroconvulsive therapy (ECT). Antidepressants alone were found to be ineffective or only partially effective in treating psychotic depression unless somatic or depressive declusions were the only psychotic symptoms. Antipsychotics alone were usually effective in providing at least a partial response, particularly with psychotic symptoms. Excellent responses of the depressive and psychotic elements were provided with ECT, ECT with antipsychotic medication, and the combination of antidepressant and antipsychotic medications. These latter treatments may be the most appropriate for depression with psychotic features.


Sleep problems and sleep medication involvement of psychiatric patients. Psychiatric patients received a computer interview that assessed 27 life problem areas. Within the sleep area, difficulty falling asleep was compared to awakening problems and appeared to be a more severe patient problem in terms of the large number of associated nonsleep problems. The patient's intensity of physical tension and degree of excessive solitary behavior were related to the problem of falling asleep. The major variables associated with awakening problems were pain and physical complications interfering with sleep. Patients with a strong sleep medication involvement were found to use multiple classes of drugs, to use a greater overall quantity of drugs, and to express various drug problems. A combination of physical tension, anxiety, and depression were predictive of patients' sleep medication involvement, whereas no one particular group of sleep problems indicated a significantly greater involvement than another group.


Tardive dyskinesia presenting as a psychosis. Tardive dyskinesia (TD), a movement disorder secondary to neuroleptic medication, is frequently found in psychiatric patients. The authors review reasons why some patients with TD go undetected. In addition, they report on another clinical situation which led to TD going undiagnosed: two cases which developed TD, then incorporated their movement disorder into a delusional system. Their clinicians focused exclusively on their psychosis, missing the underlying neurological disorder. How commonly this occurs in clinical practice is unknown.


Electroconvulsive therapy following pituitary surgery. A 39-year-old female with a history of three psychotic episodes in 6 months developed Cushingoid features and catatonia. Despite the surgical removal of a pituitary basophil adenoma and treatment with neurleptics, her catatonia worsened. Electroconvulsive therapy provided rapid, life-saving relief for this patient. Patient characteristics are discussed and treatment recommendations provided.


Preliminary characterization of bovine pineal prolactin releasing (PPRF) and release-inhibiting factors (PPIF) activity. Bovine pineal glands were subjected to extraction with dilute acetic acid, gel filtration on Sephadex G-25 and subsequent ultrafiltration through Diaflo membranes PM10, UM2 and UM05. Various fractions derived at each step were tested for the presence of substances which stimulate or inhibit prolactin secretion in vitro and in vivo. Both prolactin releasing (PPRF) and release-inhibiting (PPIF) activities were observed. PPRF activity was present in certain fractions derived from Sephadex G-25 and in the PM10 residue (MW congruent to less than 10,000). Whereas both the UM2 residue (MW greater than 1000) and UM05 filtrates (MW less than 500) was seen to inhibit pituitary prolactin release in vitro, the UM05 residue (MW greater than 500 and less than 1000) inhibited prolactin release in vivo, possibly by stimulating the secretion of the hypothalamic prolactin inhibiting factor. On the basis of its inactivation by trypsin it was concluded that PPIF may be a peptide or contain a peptide moiety indispensible for its biological activity. Experiments are in progress to characterize pineal prolactin-regulating activities and to elucidate further the physiological role of the pineal gland in the regulation of prolactin secretion.


Neurobiology of Polyorchis. II. Structure of effector systems. The gross and fine morphology of the major effector systems in the anthomedusan, Polyorchis penicillatus, is described and discussed in relation to the known physiological and behavioral properties of these systems. Swimming is controlled by an anastomosing network of giant neurons within the inner nerve ring and radial nerves. Although these neurons may be coupled by gap junctions it is likely that they form a syncytium. The photosensitivity of the "giants" is attributed to reflexive membranes within the cytoplasm. Giant neurons act as both the pre- and postsynaptic cell when forming synapses with other neurons of the inner nerve ring. Neuromuscular synapses between "giants" and the striated swimming muscle are found around the margin and along the radii. Swimming muscle cells are connected laterally by gap junctions and end-to-end by desmosomes which are sometimes elaborated with extra-thick filaments. Unstriated sphincter and radial muscles, the major muscles associated with crumpling, are both greatly folded over mesogloeal ridges and have processes that cross the mesogloea to contact the ring and radial canals, respectively. Synapses or other sites that might be responsible for exciting these muscles during crumpling have not been found. The ability of the endodermal lamella and canals to propagate action potentials can be accounted for by the numerous gap junctions that are seen in these tissues. The precise location where excitation is transferred to the nervous system to initiate crumpling is not known but epithelial bridges crossing the mesogloea are likely routes. Synapses between neurons originating in the outer nerve ring and tentacle longitudinal muscle can account for the control of tentacle length. Neurons of the outer nerve ring also synapse onto velar, radial fibers and the sphincter muscle. The inner and outer nerve rings have nervous connections. The organisation of the outer nerve ring and the arrangement of nerves within the endodermal plexus is described. A diagram showing the major connections and interactions of components of the effector systems is presented.


Acidic thiol proteinase activity of Schistosoma mansoni egg extracts. Extracts of the eggs of the human blood fluke, Schistosoma mansoni, exhibit proteolytic activity which requires the presence of added thiol reagents or cyanide. The pH optimum for hydrolysis of Azocoll and cartilage proteoglycan is 4.8--5.2 and the molecular weight of the major component is 25--26,000. The effects of inhibitors suggest this activity belongs to the acidic thiol proteinase class, with a similarity to Cathepsin B. These proteinases may be involved in nutrition of the egg or sporocyst, in penetration of eggs or miracidia through host tissues, or in the immunopathology of schistosomiasis.


Incorporation of radioactive precursors into filarial larvae of Brugia developing in susceptible and refractory mosquitoes. The incorporation of tritiated precursors injected into mosquito hosts parasitized by developing filarial larvae of Brugia patei has been studied by autoradiography in 2 species of mosquito, Aedes togoi in which filarial development was normal and Anopheles labranchiae atroparvus in which filarial development was abnormal. In both mosquito hosts there was significant incorporation into 4--5-day-old developing larvae of uridine and amino acids (isoleucine, leucine, valine, arginine, lysine, cystine, methionine, phenylalanine, tyrosine, tryptophan, histidine, and proline), although lower incorporation of methionine, tyrosine, and tryptophan was found during abnormal development. No incorporation of thymidine, hydroxytryptophan, dopa, or carbohydrate was found at this stage of larval development. Some incorporation of glucose and dopa was found in or around earlier stages of development in An. l. atroparvus. Mosquito flight muscle showed lower incorporation of glucose, but not of amino acids, around the site of filarial parasite development. The flight muscle of An. l. atroparvus showed a higher level of incorporation of lysine compared to that in A. togoi and higher levels of lysine and valine were found in the abnormally developing filarial larvae in the refractory mosquito.


The longevity and hatchability of Philophthalmus megalurus and P. gralli miracidia in different environmental conditions. The effects of salinity, pH, and temperature on the longevity and hatchability of miracidia of Philophthalmus megalurus and P. gralli were determined. Miracidia of both species are able to hatch and survive at saline concentrations much above physiological levels, although these processes are reduced in 2.0--2.4% saline and completely inhibited at 2.6%. The greatest hatching rates for both species were found near neutrality (pH 6--8) but some miracidia hatched at the extreme pH levels of 3 and 12. Philophthalmus megalurus miracidia exhibited longer half-lives under acid conditions (pH 2--6) than P. gralli miracidia; conversely, P. gralli miracidia showed longer half-lives in alkaline conditions (pH 8--11). Hatching and longevity were much greater below room temperature (5--20 C) than above (30--50 C) for miracidia of both species. Temperatures above 50 C proved lethal for eyefluke eggs. Except in acid pH, P. gralli miracidia showed longer half-lives than miracidia of P. megalurus. Comparison to studies on schistosomes revealed that the inhibitory effects of physiological saline and host body temperature on the hatching process of schistosome eggs does not occur in these 2 species of eyeflukes.


The parasitic coelenterate, Polypodium hydriforme USSOV, from the eggs of the American acipenseriform Polyodon spathula. No significant differences in macro- and micromorphology were found between the parasitic stolon and free-living polyps of Polypodium sp. obtained from infected eggs of the North American acipenseriform fish Polyodon spathula and corresponding developmental stages of Polypodium hydriforme Ussov, parasitic in the Volga sterlet (Acipenser ruthenus). Therefore, both the American and the European forms of Polypodium belong to the species P. hydriforme Ussov.


Basic study of microsurgical orchidopexy for high undescended testis; assessment of clinical applicability. An experimental microvascular surgical technique to bring an intraabdominal testis to the scrotum was used on rats and dogs. It appears that the method may be applicable to clinical cases in children.


Interactions of parenteral solutions with sulphur-treated glass bottles. Concern about limited surface durability has been the main reasons for recommendations by advisory committees and government health authorities, not to reu-use sulphur-treated soda-lime glass (type II) bottles for intravenous solutions. In order to contribute specific data, the interactions of slightly acid and neutral parenteral solutions with ammonium sulphate-treated type II glass bottles have been investigated. It was established that the amounts of silica, sodium and calcium released into the solution are not greater than the potential background contamination from the raw materials. The number of particles in the solution was well below the limits set by the British Pharmacopoieia and not much higher than the lowest background count practically achievable. On an average, bottle surfaces released less material after the first time of use. Bottle-to-bottle variations revealed by scanning electron microscopy point at problems in achieving smooth, evenly surface-treated bottle surfaces during bottle manufacture.


The assay and stability of chlorpropamide in solid dispersion with urea. Thin layer chromatography followed by reflectance densitometry has been used to evaluate the stability of chlorpropamide-urea during the fusion process. Urea was found to decompose to biuret and chlorpropamide to p-chlorobenzenesulphonamide: several other unidentified decomposition products were detected. The energy for decomposition of chlorpropamide was 57.1 kJmol-1 for melts containing 15 and 30% chlorpropamide. Decomposition followed apparent first order kinetics.


Correlation of bioavailability in man with simulated absorption data for three doxantrazole preparations. The in vitro and in vivo availability of doxantrazole, a potential antiallergic compound has been evaluated. A solution was significantly less bioavailable than either tablet or suspension formulations and it is suggested that this is associated with the large volume of the solution vehicle altering the hydrophilicity of the gastrointestinal fluids. In vitro availability was determined from absorption rate constants and absorption profiles obtained using the Sartorius absorption and solubility simulators. A statistically significant correlation was found between the percentage absorbed in vitro at 1 h and both total urinary recovery and area under plasma curve values in vivo. It is considered that in vitro determination of diffusion through artificial lipid membranes may be a useful predictive method of in vivo availability.


Serum and peritoneal fluid concentrations of clindamycin and lincomycin in the mouse. Serum concentrations of drug were obtained at various times after intramuscular dosing of healthy mice with either clindamycin or lincomycin hydrochlorides. Washings from the peritoneal cavity were taken at the same time as the serum samples. Changes in drug concentration in the peritoneal fluid with time mimic those for the serum, but concentrations in peritoneal fluid are much greater than the corresponding serum concentrations. The total volume of peritoneal fluid was estimated to be 0.05 ml.


Effects of acute or chronic administration of chlordiazepoxide on feeding parameters using two food textures in the rat. The effects of acute or chronic administration of chlordiazepoxide on several feeding parameters were studied in the rat. Given acutely, chlordiazepoxide reduced the rate of feeding, at lower dose levels (5.0 or 10.0 mg kg-1) tended to prolong feeding duration, and at a higher dose (15.0 mg kg-1) reduced the amount of food intake. Food texture can influence feeding parameters too; but the effects of the drug did not interact with the effects of food texture. After 9 daily injections of the drug, there were several indications of the development of tolerance to the effects of the drug on the feeding parameters. A possible role of 5-HT mechanisms in the reduction of eating rate produced by the drug is considered.


Monoamine oxidase inhibition and brain amine metabolism after oral treatment with toloxatone in the rat. Rats were administered toloxatone 100 mg kg-1 p.o., and killed 0.5 to 8 hours later. Toloxatone reversibly inhibited type A MAO, but did not affect the activity of type B MAO in whole brain. Cerebral concentrations of noradrenaline, dopamine and 5-hydroxytryptamine were increased after toloxatone, while their metabolite concentrations were reduced. Synaptosomal uptake processes of these amines were not altered by tolaxatone.


Selective influences of age and thyroid hormones on type A monoamine oxidase of the rat heart. The specific actiivty of rat heart MAO, towards both tyramine and benzylamine as substrates, was found to increase with the age of the animal, and also after administration of (-)-thyroxine to young male rats. Conversely, enzyme activity was decreased in animals made hypothyroid by including 2-thiouracil in their diet. However, with both age and altered thyroid status, relatively greater changes in the deamination of tyramine rather than in that of benzylamine, were obtained. Clorgyline and deprenyl, used as inhibitors of rat heart MAO, indicated that tyramine is metabolized solely by MAO-A, whereas benzylamine is a substrate for both MAO-A and -B, and also a clorgyline- and deprenyl-resistant enzymic activity. The proportional contribution of MAO-A, -B and the clorgyline-resistant enzyme towards the total benzylamine deamination in the rat heart was found to vary with the age and with altered thyroid status of the animal in such a way that selective changes in the activity of MAO-A appear to be largely responsible for the overall changes in the specific activity of rat heart MAO which occur in response to these developmental factors.


Catecholamine uptake blockade in anaesthetized dogs: influence on cardiovascular responses. The effects of differential and combined catecholamine uptake antagonism on cardiovascular responses of anaesthetized dogs to isoprenaline, noradrenaline, and electrical stimulation of the left ansa subclavia nerve have been studied. Uptake1 inhibition by cocaine HCl (5 mg kg-1 and 1 mg kg-1 every 45 min) enhanced responses to noradrenaline (0.1 to 2.0 micrograms kg-1 i.v.) and sympathetic nerve stimulation (1 to 20 Hz), but did not affect those to isoprenaline. Uptake2 inhibition by metanephrine (40 micrograms kg-1 min-1) enhanced cardiac responses to isoprenaline (0.05 to 1.0 microgram kg-1 i.v.), but did not significantly alter those to noradrenaline or nerve stimulation. Responses to all agonist interventions were increased by the combined administration of cocaine and metanephrine. Cocaine preferentially enhanced the positive chronotropic cardiac response to noradrenaline, but metanephrine did not differentiate between heart rate and contractility. These results have been discussed in the light of the mechanism of drug action involved.


Inhibitory effect of 5-hydroxytryptamine on lipogenesis in rat adipose tissues. 5-Hydroxytryptamine (5-HT) inhibited the incorporation of 14C from 14C-labelled glucose, pyruvate, citrate and acetate into fatty acids but it did not inhibit the conversion of 14C from citrate and acetate into CO2, and the citrate conversion into glyceride-glycerol in epididymal and mesenteric adipose tissue from 24h-fasted rats. 5-HT stimulated the formation of lactate from glucose and pyruvate, and increased the ratio of lactate produced/pyruvate taken up. This ratio was similar to the NADH:NAD ratio. These results indicate that 5-HT inhibits fatty acid synthesis in rat white adipose tissue by mechanisms similar to those of the catecholamines.


A method to predict infinity values for biexponential processes. An equation is presented which allows infinity values for biexponential processes to be predicted in the early nonlinear phase when samples are taken at equal time intervals. This equation is independent of the value or ratio of the rate constants involved in the process. However, this method is very sensitive to noise normally associated with urine data.


Degradation of mecillinam in aqueous solution. The hydrolysis of mecillinam in aqueous solution (37 degrees) was studied at pH 2-10. The degradation products observed by TLC and NMR were identified and quantified. Several of these compounds were synthesized. Mecillinam and the key degradation product, (6R)-6-formamidopenicillanic acid, underwent reversible 6-epimerization in basic solution. Some of the thiazolidine derivatives formed epimerized at position 2. In contrast to penicillins, the degradation pattern of mecillinam becomes more complex with increasing pH. Rate constants for some processes are given.


New and simple methylene blue colorimetric assay for glycyrrhizin in pharmaceuticals. A new colorimetric method for glycyrrhizin in licorice and drug preparations is given. The method is based on coupling the acidic genin, glycyrrhetinic acid, with methylene blue and measuring the extinction of the coupled compound solution in chloroform-alcohol.


Synthesis and pharmacology of potential beta-blockers. Several 1-(4-substituted phenoxyl)-2-hydroxy-3-isopropylaminopropanes and 1-(4-substituted phenoxy)-2-hydroxy-3-[3,4-dimethoxyphenethyl]aminopropanes were synthesized for possible beta-adrenergic receptor blockage. The compounds were synthesized by reaction of the 4-substituted phenol with epichlorohydrin and subsequent opening of the resulting epoxide with either N-isopropylamine or N-3,4-dimethoxyphenethylamine. Preliminary biological testing indicated a decrease in the beta-blocking potency and the duration of action.


Urinary excretion of chlorpheniramine and pseudoephedrine in humans. A specific high-pressure liquid chromatographic method for the determination of chlorpheniramine and pseudoephedrine in urine was developed and applied in a urinary excretion study of normal healthy subjects who received a sustained-release dosage form contianing 8 mgof chlorpheniramine maleate and 120 mg of pseudoephedrine hydrochloride. Five subjects received one dose on Day 1, followed by multiple dosing every 12 hr for 7 days without ammonium chloride administration. Four subjects received one dose of the sustained-release dosage form together with ammonium chloride. Urine samples were collected during the 1st day and at steady state. The method is specific and simultaneously determines choorpheniramine, two metabolites (mono- and di-desmethylchlorpheniramine), pseudoephedrine, and norpseudoephedrine. The assay recovery was less than 97% (0.06-3 microgram/ml) for chlorpheniramine maleate and less than 98% (1.5-75 microgram/ml) for pseudoephedrine hydrochloride. Excretion of chlorpheniramine and its two metabolites in urine was enhanced after ammonium chloride administration. At steady state, a change in urine pH from 5.69 to 6.46 resulted in more than a 25% decrease in chlorpheniramine and monodesmethylchlorpheniramine excretion. In spite of expected changes in its biological half-life, the overall amount of unchanged pseudoephedrine excreted in urine was not affected by urine pH, presumably because it is primarily excreted in urine as intact drug.


Anomalous solution behavior of 2-palmitate esters of lincomycin and clindamycin. The aqueous solubilities of lincomycin and clindamycin 2-palmitate esters are compared. Clindamycin 2-palmitate hydrochloride has an unusually high solubility at 25 degrees, which is due to micelle formation. Both compounds are surface active with relatively low critical micelle concentrations. However, since the Krafft point of lincomycin palmitate is approximately 43 degrees, it does not form micelles below  that temperature and appears to be quite insoluble until heated above 43 degrees. The experimental monomeric solubilities  of the two compounds agree with calculations based on group contributions to lipophilicity. Clindamycin 2-palmitate hydrochloride solutions are quite sensitive to ions, being salted out as unprotonated base in the form of oily droplets. Salting out correlates well with anionic strength, which is quite constant for the various salts studied. A viscosity maximum occurs with increasing salt addition, with the peaks of the different salts occurring at the same anionic strengths.


Plasma binding of benzodiazepines in humans. Plasma binding of chlordiazepoxide, diazepam, loraxepam, and oxazepam was determined by equilibrium dialysis in 20 male, healthy volunteers, 25-86 years old. A wide range of binding was observed, with the free fraction varying twofold for lorazepam, fourfold for chlordiazepoxide and diazepam, and over 20-fold  for oxazepam. Statistically significant linear relationships were not observed between the degree of binding and age, serum albumin, or total protein for any of the drugs. There was, however, a correlation between the extent of binding for the four drugs. Because of the importance of unbound benzodiazepine levels in eliciting any pharmacological response and also in disposition, consideration of the wide interindividual variability in plasma binding must be made in interpreting pharmacodynamic and pharmacokinetic data.


Capacity-limited gut wall metabolism of 5-aminosalicylic acid, a therapeutically active metabolite of sulfasalazine, in rats. The metabolic fate of 5-aminosalicylic acid (reported to be the active therapeutic moiety of sulfasalazine) was assessed in fasting rats as a function of dose (25-200 mg/kg) and administration route (oral, intraperitoneal, and intravenous). 5-Aminosalicylic acid is subject to both capacity-limited presystemic (apparently during first passage through the intestinal epithelium) and systemic acetylation. The possibility exists that 5-aminosalicylic acid also is acetylated presystemically after oral sulfasalazine administration to patients with inflammatory bowel disease. Any alteration in the absorption activity if N-acetyl-5-aminosalicylic acid is inactive or less active than 5-amino-salicylic acid.


Degradation kinetics of a new cephalosporin derivative in aqueous solution. The degradation kinetics of a new cephalosporin derivative (1) in aqueous solution were investigated at 60 degrees, mu = 0.05, at pH 2.0-10.0. The observed degradation rates followed pseudo-first-order kinetics and were influenced significantly by H2O and OH- catalysis. No primary salt effect was observed in the acid region, but a positive salt effect was observed at pH 9.4. A general base catalytic effect by a phosphate buffer species was observed at pH 7-8. The pH-rate profile for I exhibited a degradation minimum at pH 6.05. The Arrhenius activation energies determined at pH 4.0 and 9.4 were 27.2 and 24.5 kcal/mole, respectively. Excellent agreement between the theoretical pH-rate profile and the experimental data supported the hypothesized degradation process. A comparison of I and cefazolin revealed close structural and stability analogies.


Acidosis inhibits the hypocalcemic effect of acetazolamide. The effect of acetazolamide on calcium metabolism was examined using sham-operated, ureter-ligated and nephrectomized rats. Acetazolamide doses from 10 to 500 mg/kg produced significant hypocalcemic effects in ureter-ligated and nephrectomized rats. However, doses of acetazolamide up to 1000 mg/kg were devoid of hypocalcemic activity when administered to sham-operated rats. Sham-operated rats exhibited an acidotic response to acetazolamide while ureter-ligated rats did not. Attenuation of this drug-induced acidotic response with i.p. injections of tris(hydroxymethyl)amino-methane uncovered a hypocalcemic effect of acetazolamide in sham-operated rats. Also, the hypocalcemia associated with acetazolamide treatment of ureter-ligated rats was negated when an acidosis was induced by prior injection of NH4Cl. These data indicate that the administration of inhibitors of carbonic anhydrase produces a hypocalcemia when a metabolic acidosis is not present.