[The value of dobutamine in cardiology]. The hemodynamic effects of dobutamine have been evaluated by catheterization for a limited duration in 17 patients. The cardiac index, the systolic index, the myocardial contractility given by the values dp/dt and Vmax, and the systolic work index have increased significantly while the increase in the mean aortic pressure and the cardiac rate were small. The left ventricular telediastolic pressure, and systemic and pulmonary arterial resistance diminished significantly. These results seen to authorise the prolonged use of this drug in the low output syndromes with left ventricular failure whether the cause be ischemic or due to a cardiomyopathy. Nine patients were then treated, with a dosage of 8 microgram/kg/min. The supervision of the treatment was composed of iterative readings of the arterial pressure, the cardiac rate, pulmonary capillary and arterial pressures, and the cardiac index. The hemodynamic results were similar to those obtained in the 17 patients. It seems that this product can be used in the acute stage of myocardial infarction. Indeed its action on the myocardial excitability is small and the moderate increase in the oxygen consumption can be compensated for by the improvement in the myocardial vascularisation.


[The effects of dobutamine in postoperative disorders of left ventricular function in coronary patients undergoing abdominal surgery. Apropos of 18 cases]. Increasing doses of dobutamine were administered to the first group of 10 coronarians having undergone and abdominal surgical procedure, and presenting, one hour after awakening from the anesthesia, hemodynamic modifications with a diminution of cardiac index (CI), systolic index (SI), systolic work index of the left ventricle (SWILV) increase in the pulmonary capillary pressure (PCP), and in the total peripheral resistance (TPR), as well as an acceleration of the cardiac rate (CR). Doses of dobutamine of 5 or 7.5 microgram.kg-1.min-1 corrected the IC, PCP and TPR. Dobutamine ameliorated the SI and SWILV in an increasing fashion up to a dose of 10 microgram.kg-1.min-1 only and without restoring them to the control values of the pre-operative period. CR progressively increased with the increasing of the doses reaching 126 +/- 21.5 beats min-1 for 15 microgram.kg-1.min-1. Extrasystoles appeared at dose levels of 12.5 and 15 microgram.kg-1.min-1 in two patients. Tests of vascular filling (pre-charge tests) carried out in the second group of patients under 10 microgram,kg-1.min-1 of dobutamine and in a third group under 15 microgram.kg-1.min-1 showed a good cardiac adaptation to filling, equal or superior to that of the pre-operative period. It also appeared that the amelioration of CF obtained with a moderate vascular filling (300 ml of low molecular weight dextran) under 10 microgram.kg-1.min-1 of dobutamine is greatly superior to the amelioration obtained by 10 to 15 microgram.kg-1.min-1 of dobutamine.


[Hemodynamic effects of dopamine, dobutamine and isoproterenol in chronic respiratory insufficiency]. A study of the effects of dopamine, dobutamine and of isoprenaline was carried out in 20 patients with chronic respiratory insufficiency, 16 of whom had pulmonary arterial hypertension (PAP: 22 mmHg). The following findings emerged: --absence of any significant effect of dobutamine on heart rate,--increase in systolic and mean PAP, after infusion of dobutamine,--increase in systolic and mean PAP, after infusion of dopamine,--decrease in BP after administration of infusion of dobutamine,--decrease in pulmonary arterial resistances under the influence of isoprenaline,--decrease in total systemic resistances after the administration of dobutamine,--increase in ventricular, systolic work after the infusion of dopamine.


[Bacteriological studies of peritonitis]. Bacterial infections of the peritoneum may be primary (pneumococcal peritonitis of childhood). These are rare, as are Streptococcal and Staphylococcal peritonitis in the neonate, following umbilical or tegumental infections. The great majority of cases of peritonitis are secondary to an adjacent inflammation, a perforation of the gastro-intestinal tract, or trauma, which may be surgical in origin. The microbial flora involved is generally mixed, with a high proportion of anaerobic organisms, reflecting the composition of the intestinal flora, and in particular, that of the colon. Infections due to a single organisms are less common, and are seen principally in superinfected ascites, high perforations, or following preoperative antibiotic therapy. The treatment of peritonitis is thus dependent upon the microbial flora present. In mixed infections, broad spectrum antibiotic therapy (amino-glycosides + beta lactamines) is necessary, combined with a b-nitro-ionidazole derivative in view of the frequent presence of Bacteroides fragilis. When the infection is due to one or a limited number of organisms, only the antibiotic sensitivy will provide useful indications. The dangers of preoperative "sterilization", which leads only to the selection of multiresistant strains, is emphasized. Such sterilization is thus to be proscribed in the prevention of peritonitis. Antibiotic cover, on the other hand, is still indicated.


[Value of and indications for noxythiolin in generalized peritonitis]. The favourable experience of M. BROWN with noxythiolin in fecal peritonitis has prompted us, over the last seven years, to use this product in all forms of generalized peritonitis. This study involved 187 cases of generalized peritonitis : 102 males and 85 females, whose mean age was 53 years. Noxythiolin was reserved for severe cases (after having deliberately decided not to perform a comparative control series), the severity being related to the advanced age of the patients, the delay in intervention, and preexisting disorders. It was used in the form of a peroperative lavage of the peritoneal cavity and a postoperative instillation. The overall survival (71 p. 100) and that observed in particular in peritonitis of colonic origin (58 p. 100), seem to be arguments in favour of the beneficial effect of the product. Noxythiolin contributes to the sterilization of the peritoneal cavity. By virtue of its efficacy, the simplicity of its use, and its innocuousness, it merits a place in the therapeutic arsenal for treatment of generalized peritonitis, in combination with systemic treatments.


[Results of noxythiolin use in acute peritonitis]. The authors present a series of 30 cases of acute supra- and inframesocolic peritonitis which, in addition to the treatment of their organic causes, benefitted from medical treatment by noxythioline, used as a peritoneal lavage, an irrigation with aspiration, or an instillation. A marked amelioration in the postoperative course was noted, particularly in those cases with infra-mesocolic peritonitis, with the absence of adhesions and suppurative collections on reintervention, and a rapid return to negativity of the positive bacteriological investigations performed systematically on the pe ritoneal fluid and on the drainage.


[Postoperative peritoneal irrigation in generalized peritonitis treated in an intensive care unit]. Over a period of four years, 16 patients with generalized peritonitis have been treated by postoperative peritoneal irrigation in an Intensive Care Unit. The majority of cases involved postoperative peritonitis accompanied by severe visceral failure. The irrigation liquid, containing an antibacterial agent, was perfused for between one and eleven days at a high flow rate (mean 16.5 I). The overall mortality in the series was extremely high (75 p. 100), and even greater (85 p. 100) if only the cases of postoperative peritonitis were considered. Anatomical examination of the peritoneum, performed on reintervention or at autopsy, revealed an abnormally high incidence of residual abscesses. These are responsable for the continuation or recurrence of infection, and explain the high mortality. These observations, combined with the frequent occurrence of local or general complications, led the authors to reject peritoneal irrigation in postoperative peritonitis in the presence of severe visceral failure, and to use it, in selected cases, only for periods not exceeding 48 hours.


[Antibiotic treatment in acute peritonitis]. The selection depends on several factors and in particular: -the means of transport by which the antibiotic reaches the peritoneum : the aminosides and the betalactamines attain the peritoneum easily while the polymyxins do not;  -the causative agent: most often it is a gram negative aerobic or anaerobic bacteria. The entero-bacteria predominate but in the last few years Bacteroides fragilis has been frequently encountered giving rise to numerous studies. The flora is most often polymicrobic. The betalactamines act on gram positive bacteria whether they are aerobic or anaerobic. The aminosides are active against aerobic gram negative bacteria. The same can be said for Colimycine (except for certain Proteus and Providencia). All these antibiotics (other than carbenicillin) are not effective against Bacteroides fragilis which are however very sensitive to lincomycine, to the cyclines and in particular to doxycycline and to the derivatives of imidazole. The laboratory is an important aid in judging therapeutic effectiveness by determining the bacteriacidal strength of the serum and even the serum level of the antibiotic whatever the cause it is imperative: to avoid toxicity and to pay particular attention to the frequent problem of renal insufficiency in these subjects.


[Focalized irrigation-lavage and sequential use of a new antiseptic by local and general route. Preliminary report apropos of 2 cases of suppurative pancreatic necrosis]. In view of the severe course seen in the presence of any suppurated pancreatic necrosis, it was felt to be of value to treat two patients by the adjuvant use of a new antiseptic tauroline, administered locally and, where appropriate, systemically. After surgical excision of pancreatic and peripancreatic necrotic tissue, the authors prefer to avoid active suction drainage of the residual pocket, replacing it by continuous and prolonged post-operative irrigation-lavage, together with the sequential, twice-daily use of a solution of 2 p. 100 tauroline. In view of the presence of a Gram negative septicemia (organisms identical to those of the suppurated area), the same substance was given by intravenous infusion in a 0.5 p. 100 solution, this being the only systemic, anti-infectious therapy. The effectiveness, good tolerance and originality of this new substance, active against Gram negative, Gram positive, aerobic and anaerobic organisms as well as yeast, have led the authors to immediately report their initial experience with its use.


[Purulent pleurisy caused by anaerobic bacteria. Apropos of 2 observations]. Purulent pleural effusions due to anaerobic organisms secondary to infections of the pulmonary parenchyma are complications which often occur following the inhalation of organisms from the buccal cavity. The most commonly found organisms belong to the endogenous anaerobic flora of Veillon. Rapid encystment of the pleural collection is suggestive of this diagnosis. Treatment consists of drainage of the cavity associated with specific antibiotics.


[Use of a new solution of synthetic amino acids in postoperative surgical intensive care. Evaluation of the value of nitrogen intake started in the immediate preoperative period]. A new synthetic amino acid solution, Totamine, was studied in pre- and postoperative surgical intensive care in order to determine the possible value of beginning high protein intake during the preoperative period. A comparison was made between two groups of 11 and 15 patients respectively, divided on a strictly random basis, receiving 1200 calories and 12.4 g of nitrogen per day in the forme of Totamine carbohydrate concentrate, starting two days before operation in one of them and on the day of surgery in the other. No statistically significant difference was seen. Calorific and nitrogen intake in both cases appeared to be satisfactory during the early preoperative period and both local and general tolerance of the amino acid solution was excellent. During the late postoperative period, there was a tendency to negativisation of nitrogen balance which justified an increase in calorific and nitrogen intake.


[Acute renal insufficiency caused by phenyl-indane-dione. Apropos of 1 case]. One case of Phenindione (PID) adverse reaction is reported. The patient showed a typical picture of immunological reaction to the drug. In spite of severe bacteremia, she recovered. Only 33 cases of PID intolerance are reported in the literature. In all these patients, renal failure occurred. Superinfection is the most frequent cause of death. PID adverse reaction should be evoqued in the presence of signs such a fever, asthenia, anorexia and cutaneous reaction. The PID should be stopped immediatly but renal failure yet develops. During a PID treatment, frequent evaluation of blood azotemia, creatinine and proteinuria should be performed.


Development and prospective study of an Anatomical Index and an Acute Trauma Index. The development of two indices which can be used to characterize the acutely injured patient is presented. Evidence in support of the concept of characterizing injuries by the combined use of tissue disruption information and physiologic and biochemical response information is examined.


Changes in hydromineral metabolism in diarrhoeic rabbits. 2. Study of the modifications of electrolyte metabolism. The study of the equilibrium of the main electrolytes (Na+, K+, Cl-), in plasma and intestinal content and the evolution of the partial plasma (Na+, K+, Cl-) in the young rabbit suffering from diarrhoea revealed disturbances of the mineral metabolism. Mineral losses were lower in sick animals than in controls. When the reduction of ingested matter was taken into account, the ion balance showed a deficit. The apparent coefficient of digestive utlization (aCDU) of the electrolytes Na+ and K+ dropped, although the reabsorption of these ions in the last segments of the digestive tract was generally good. The pH remained normal and the proteinaemia fell, suggesting a haemodilution. This was backed up by the presence of a hyponatraemia and a hypochloraemia, but did not account for the large drop in kaliemia. On the other hand, the plasma osmotic pressure increased, very probably in association with a high uraemia. It would therefore seem that the pathogenesis of diarrhoea in the young rabbit differed from that of other known examples, such as diarrhoea in the calf and in the infant. In the young rabbit, the pathogenesis of diarrhoea resulting from coccidiosis would seem to be, as much a result of a nutritional deficiency in particular of potassium, as of a malfunctioning of the digestive tract.


Optimizing the continuous production of Candida utilis and Saccharomycopsis fibuliger on potato processing wastewater. The yeasts Candida utilis and Saccharomycopsis fibuliger were propagated as a source of single-cell protein in a continuous, mixed, aerobic, single-stage cultivation on blancher water generated during potato processing. A series of steady-state experiments based on a two-level factorial design, half-replicate modified with an intermediate experiment, was performed to determine the effect of pH, 3.8 to 4.8; dissolved oxygen, 42 to 80% saturation; dilution rate, 0.17 to 0.31 h(-1); and temperature, 27 to 32 degrees C on the amount of carbon consumed, the rate of carbon consumption (R(c)), the amount of reducing sugar consumed, the rate of sugar consumption (R(g)), the amount of protein produced, the rate of protein production (R(p)), the yield from carbon, and the yield from reducing sugar. The results were analyzed by stepwise multiple regression and Fisher's least significant difference test. Analyses showed that high dilution rates resulted in increased R(c), R(g), and R(p) and indicated that a rate of 0.31 h(-1) was below the critical dilution rate. A temperature of 32 degrees C increased the amount of carbon consumed by 34%. A pH of 4.3 to 4.8 increased the amount of protein produced. The yield from carbon was constant, and the relatively high yield from reducing sugar indicated that other substrates were consumed. Dissolved oxygen was in excess at 42% saturation and above. Since C. utilis predominated the mixed cultures and amylase production appeared to be limited, a single-stage fermentation lacked efficiency. The experimental design allowed preliminary optimization of major environmental variables with relatively few experiments and provided a basis for future kinetic studies.


Injury and recovery of Escherichia coli after sublethal acidification. Over 99% of the viable cells of Escherichia coli K-12 were injured after a 60-min exposure to 0.3 M sodium acetate buffer at pH 4.2. Injured cells were those able to grow on Trypticase soy agar but unable to grow on violet red bile agar. The extent of both death and injury of acid-treated cells increased with decreasing pH or increasing concentration of acid. Injured cells were able to recover their colony-forming ability on violet red bile agar by incubation in Trypticase soy broth or potassium phosphate buffer before plating on the agar media. Direct neutralization of the injury medium did not allow recovery and, in fact, was lethal to the population. The addition of metabolic inhibitors to the Trypticase soy recovery broth was used to study the repair process. It was not affected by the presence of inhibitors of protein, cell wall, deoxyribonucleic acid, or ribonucleic acid syntheses. The addition of 2,4-dinitrophenol to the recovery medium also did not inhibit recovery. Actinomycin D and N,N'-dicyclohexylcarbodiimide were lethal to a proportion of the acidified cells but allowed another fraction of the population to recover. There were no detectable amounts of 260- or 280-nm-absorbing materials leaked during the course of acid injury.


Formation of N-nitrosamines from seconday animes and nitrite by resting cells of Escherichia coli B. In the presence of resting cells of Escherichia coli B, the formation of N-nitrosamines from nitrite and secondary amines, such as dimethylamine and piperidine, was proportional to the incubation time and to the cell concentration. Optimum pH was 8.0. Boiled cells were incapable of nitrosating secondary amines. Although these experiments were carried out by using intact cells of E. coli B, the reaction followed Michaelis-Menten kinetics, and the apparent Km values calculated from Lineweaver-Burk plots were 0.12 +/- 0.03 M for dimethylamine and 0.07 +/- 0.02 M for nitrite. The apparent Km for piperidine was 0.15 +/- 0.05 M. The nitrosation was inhibited by high substrate concentrations. These results suggested that the formation of n-nitrosamines by resting cells of E. coli B apparently depends on their enzyme activities.


Relationship between arterial and venous bicarbonate values. We determined the clinical efficacy of using the venous CO2 value, as obtained with routine "electrolytes", in acid-base management. Venipuncture samples for venous CO2 content and chloride concentrations were obtained in 336 patients with arterial blood pH, PaO2, PaCO2, and oxygen saturation determinations. The linear correlation between actual calculated arterial HCO3- and the measured venous CO2 was significant (P less than .001). Using venous CO2, chloride, and arterial pH values, we present various prediction equations for estimating arterial HCO3-. We determined the effects of time delay, exposure to air, and acute changes in arterial blood pH and PaCO2 on venous CO2 levels. Venous CO2 determinations should not be substituted for the arterial HCO3 value in the Henderson-Hasselbalch equation to calculate arterial blood pH or PaCO2. Clinically, the venous CO2 value has little direct use, but when venous CO2 content is abnormal, it should alert the clinician to the need for obtaining arterial blood gas and pH values.


[Effects of beta blockaders on ventilatory function in chronic bronchitis]. This study was carried out to determine whether beta blockers could be prescribed for patients with coronary insufficiency and chronic bronchitis. The effects of intravenous infusions (30 mn) of propranolol (30 micrograms/kg), practolol 90 micrograms/kg), atenolol (90 micrograms/kg) and acebutolol (150 micrograms/kg) on vital capacity and expiratory flow rates were investigated in chronic bronchitics. Propranolol (n = 51) moderately reduced the vital capacity and FEV1, by an average 9% and a maximum of 20%. The three other infused agents given to groups of 10 patients did not change the ventilatory function. When the same patients were investigated by cross over with propranol bronchoconstriction was observed. This effect was seen in all stages of chronic bronchitis but was much less severe than in a group of 50 asthmatic patients (-23%). The respiratory tolerance of the cardioselective beta blockers seems to be better but there is considerable individual variation and the diagnosis between asthma and chronic bronchitis may itself be very difficult.


Blood levels of neuroleptic drugs in nonresponding chronic schizophrenic patients. Blood levels of butaperazine were measured in schizophrenic patients who were chronic nonresponders to their psychotropic medication. The blood levels were compared with those in patients who had shown a better clinical response to this neuroleptic. Nonresponders had two to seven times lower levels of butaperazine in plasma and RBCs after a single dose or chronic dosing. Some of the patients later treated with thioridazine or haloperidol had lower plasma levels of these neuroleptics also. No significant differences were found between nonresponders and relative responders in either the alpha- or beta-phase half-life of butaperazine in plasma and RBCs after administration of a single dose of the drug. Butaperazine and thioridazine levels were not related to previously administered amounts of neuroleptic drugs. These findings do not support the hypothesis that low blood levels are the result of faster systemic metabolism of the drug after it reaches the central circulation. Our results suggest that low blood levels of neuroleptics may be one important factor in the poor clinical response of some chronic schizophrenic patients.


Tardive dyskinesia--reversible and persistent. Twenty-one hospitalized patients over 50 years of age who had tardive dyskinesia were studied for 13 months. After withdrawal of neuroleptics and antidepressants for three months, dyskinetic symptoms abated in 12 patients and persisted in nine. Discriminant function analysis showed that the persistent and reversible dyskinesia groups could be clearly separated and that the best discriminator was the number of drug-free intervals. The persistent dyskinesia group had had significantly longer neuroleptic treatment (mean, 10.8 yr) and a greater number (mean, 5.6) of drug interruptions of at least two months' duration each than did the reversible dyskinesia group. Our finding, as well as the literature reviewed, does not support the commonly held notion that frequent lengthy interruptions of long-term drug treatment reduce the incidence of persistent dyskinesia, at least in patients who are otherwise predisposed to the development of tardive dyskinesia.


Improvement in tardive dyskinesia after muscimol therapy. Muscimol, thought to be a agonist of gamma-aminobutyric acid (GABA), was administered to eight neuroleptic-free subjects with tardive dyskinesia. At oral dose levels from 5 to 9 mg, involuntary movements were consistently attenuated, usually in the absence of sedation. These results support the view that pharmacologic attempts to stimulate GABA-mediated synaptic transmission may afford symptomatic relief to patients with tardive dyskinesia.


Relationship between the B-mitten EEG pattern and tardive dyskinesia. A pilot control study. The identification of patients at high risk for the development of tardive dyskinesia (TD) is a major problem in psychopharmacology. We found a possible relationship between the subcortical B-mitten EEG pattern and TD. Twenty-one TD pateints were matched on a number of relevant variables with 21 patients without TD. Ninety-five percent of the TD patients had mittens, compared to 33.3% of controls. The results suggest that the mitten dysrhythmia may be a risk factor for the development of TD. Additional findings suggest that among TD patients, mittens are differentially more frequent in younger (93.1%) as opposed to older (0.0%) subjects.


A rabbit used as a model for testing hematopoietic tissue transplants. Allogenic transplantations of bone marrow or spleen cells were carried out on rabbits treated with cyclophosphamide. About 50% of the recipients developed chimerism of peripheral blood cells. Chimerism was a passing phenomenon prevailing from 14 to 180 days after grafting. A phenomenon indicating that not all cell lines develop after transplantation from the donor stem cells, was observed. This phenomenon has been called "restricted chimerism". The GvH reaction was of a chronic type. Relative spleen weights were significantly increased in animals receiving the spleen cell grafts, but not in animals receiving bone marrow grafts. A direct positive erythrocyte Coombs test was found in almost all graft recipients. In addition cytotoxic autoantibodies against lymphocytes were present in sera of about 20% of the recipients.


Enhancing effect of X-ray irradiation on a new blood vessel formation in mice tested by lymphocyte induced angiogenesis assay. X-irradiation enhanced a new blood vessel formation occurring in the course of a local GvH reaction in mice. Maximal enhancing effect was exerted by single total body irradiation doses ranging from 700 to 1050 R. Fractionation in daily doses smaller than 200 R did not exhibit enhancing effect. Proliferative ability of angiogenesis inducing cells is not necessary for enhancing effect of irradiation.


Inhibition of lymphocyte-induced angiogenesis by enzymatically isolated rabbit cornea cells. Corneal and kidney cells were isolated from adult rabbits by enzymatic digestion. The were tested for anti-angiogeneic activity by the lymphocyte-induced angiogenesis assay. In this assay an intradermal injection of semi-allogeneic lymphocytes resulted in a new blood vessel formation visible after three days at the injection site. Isolated rabbit cells were mixed in 1:10 ratio with murine lymphocytes and injected into 600 R X-ray irradiated mice. Number of newly formed blood vessels evoked by lymphocytes injected alone or with rabbit cells added was counted. Corneal cells but not kidney cells decreased angiogenesis evoked by lymphocytes. This finding is discussed in view of corneal avascularity and pathological neo-vascularization.


Local graft versus host reaction in the xenogeneic system. The injection of viable rat spleen cells beneath the renal capsule of cyclophosphamide (CY)-treated adult mice produced a localized graft versus host reaction (GvHR), manifested by the kidney enlargement index (KI). The optimal xenogeneic GvHR was obtained after injection of 30-50 million rat spleen cells into mouse recipients treated 24 hr before with CY, 200 mg/kg intraperitoneally (i.p.), and killed 7 days later. Splenomegaly in recipient mice suggested systemic dissemination of the local GvHR.


Splenic suppressor cells in fetal and newborn mice. Newborn mice are refractory to sensitization with picryl chloride and the graft versus host reaction (GvHR) in F1 fetal mice injected with parental lymphocytes is markedly reduced. Moreover, spleen cells (but not thymic lymphocytes) of fetal or newborn animals are able to suppress the cell-mediated immune responses, contact sensitivity (CS) and GvHR in a variety of experimental situations. The inhibitory cells are thy-positive and their suppressive activity wanes early after birth.


Alkali-stable oligosaccharides isolated by hydrazinolysis of M and N blood group glycoproteins. Three alkali-stable oligosaccharides free of amino acids were purified from hydrazinolysis products of M or N blood group glycoprotein. These three oligosaccharides differed in the content of sialic acid and contained similar amounts of other carbohydrate components. The asparagine-linked oligosaccharides, obtained by strong alkaline degradation of M glycoprotein, showed similar electrophoretic mobilities and chemical composition to the oligosaccharides obtained by hydrazinolysis. It indicates that hydrazinolysis does not give a substantial degradation of carbohydrates of alkali-stable oligosaccharides of human red cell glycoprotein.


Physicochemical properties of a proline-rich polypeptide (PRP) from ovine colostrum. Properties of a proline-rich polypeptide (PRP) accompanying ovine colostral IgG2 are described. PRP is soluble at 4 degrees C but reversibly precipitates by warming to room temperature. Maximal precipitation is observed at pH = 4.6, temp. 48 degrees C, and ionic strength higher than 0.6. There is a linear dependence of precipitation on concentration of PRP. Molecular weight of PRP is 38,000 daltons. It is not changed in the presence of 6 M guanidine hydrochloride, SH-compounds, and in the presence or absence of metal ions. PRP is built of one polypeptide chain. No difference in proteolysis of IgG2 by pepsin, papain and trypsin in the absence or presence of PRP was found.


Modulation of human T lymphocyte rosette formation by serum factors. It was determined that in the serum of TB patients, the factor (beta-lipoprotein) was present, lowering/modulating the ability of T lymphocyte rosette formation with SRBC. It was also found that in normal sera a factor is present which restores rosette formation with T lymphocytes from TB patients. The reaction of rosette inhibition factor and rosette restoring factor with xenogeneic cells, suggest that the reaction of the factors is not limited to modulation of spontaneous rosette formation only.


Antigenic and immunogenic properties of gamma-glutamyl-transferase preparations. Incubation time of human kidney gamma-glutamyl-transferase (GGT) with specific antiserum and precipitate formation caused no effect on inhibition of the enzyme activity. In the obtained precipitate GGT activity was independent of the amount of antiserum and contained 30% of the initial enzyme activity. After reaction of the serum with human liver GGT or with the urine enzyme a precipitate formation and decrease in the enzyme activity was noted. In the serum incubated with GGT isolated from animal kidneys, neither inhibition nor precipitation was observed. The precipitate of bovine kidney GGT with a specific antibody independently of the amount of antibody contained 50% of activity. After treatment of bovine GGT with bromelain four protein fractions giving precipitin lines in crossed-immunoelectrophoresis were observed but only one of them preserved catalytic activity. Immunogenicity of newly obtained immobilized GGT preparations was much lower than that of the native enzyme. A distinct and different influence of antibody on the affinity of native and immobilized GGT preparations to donor and acceptors of gamma-glutamyl group was noted. The immunofluorescent technique was used for GGT localization in some bovine tissue sections.


Synthetic antigens. Horse "natural" antibodies against interpolymer of styrene and maleic acid (PSM). Properties of horse natural anti-PSM antibodies are described. The antibodies were of IgG class. Electrostatic forces were mainly involved in reaction of PSM with horse antibodies. The reaction was inhibited by low molecular compounds resembling structural unit of PSM. Studies of difference spectra and ORD and CD spectra showed no major conformational changes in horse antibodies after reaction with PSM.


Synthesis of new sulfuric derivatives of allobarbital (5,5-diallylbarbituric acid) with anti-inflammatory activity. It was found that allyl group of alkenes Ia-Ig is transformed into 3'-sulfo-2'-sulfatopropyl or beta-methyl-beta-sulton groups in reaction of addition under action of concentrated sulfuric acid in the presence of methyl cyanide. It was stated that this reaction is competitive to the Ritter reaction. Soluble in water calcium salts IV obtained from compounds Ia and Ib exhibited strong antiinflammatory activity.


Subendocardial protection during cardiopulmonary bypass. Its use with methylprednisolone and glucose-insulin-potassium. Fifteen adult dogs were equally divided into three groups: group 1 (control) received no medication before or after cardiopulmonary bypass (CPB), group 2 was administered glucose-insulin-potassium (GIK) according to Maroko's protocol before, during, and after CPB, whereas group 3 was treated with methylprednisolone (30 mg/kg) 30 minutes prior to and near the end of CPB. Moderately severe subendocardial hemorrhage was present in 80% of group 1, 50% of group 2, and only 20% of group 3 ventricles. Endocardial viability ratios after CPB were greater than 0.75 in 20% of group 1, 80% of group 2, and 100% of group 3 animals. Both GIK and steroids improved subendocardial perfusion after anoxic arrest and CPB compared with controls. However, steroid-treated dogs exhibited less endocardial hemorrhage, improved contractility, and more rapid return to baseline conditions than either GIK or control animals. These data suggests that steroids could be considered whenever normothermic anoxic cardiac arrest is used for cardiac operations.


Failure of cimetidine prophylaxis in the critically ill. Stress ulcers with subsequent hemorrhage are a well-recognized cause of morbidity and mortality in patients hospitalized for other serious illnesses. In 39 critically ill patients, cimetidine was used as a prophylaxis against stress ulcers with the intent to keep the gastric pH level above 4. In 11 of the 39 patients, cimetidine alone did not effect consistent elevation of the gastric pH level above 4. Of the 11 patients, nine had positive blood cultures or clinical infection and five also developed renal failure. Five comatose patients never had pH greater than 4 on cimetidine alone. The mortality for those whose conditions failed to respond to cimetidine was five times greater than for those whose conditions responded to cimetidine. These data imply that a high-risk group of patients can be identified whose conditions may not respond to cimetidine treatment and whose gastric pH must be regularly monitored.


Evaluation of intragastric pH in acutely ill patients. The effect of various modalities on maintaining a high intragastric pH in acutely ill patients was evaluated. Twelve patients with one or more organ system failures had the effect of nasogastric suction, intragastric antacid instillation, and intravenous cimetidine administration on intragastric pH determined by an indwelling, intragastric pH probe. Each therapeutic modality was administered for 12 hours and the order of performance randomized. Nasogastric suction was associated with a constant intragastric pH of less than 2.0. Mean intragastric pH with cimetidine administration was significantly higher than with antacid administration and consistently greater than 5.0. If low intragastric pH represents susceptibility to acute mucosal lesions, cimetidine therapy was more effective than antacids in the doses and frequency of administration used in this study in maintaining a high intragastric pH, and it may be effective in preventing stress ulcer formation.


[Morphohistochemical characteristics of structures of the esophageal cross-striated muscle tissue]. On the bases of histochemical enzymatic studies and morphometric investigations, some differences were revealed between cross-striated esophageal muscular tissue and other skeletal muscles of mammals. A peculiar feature of the esophageal musculature is prevalence of unitypical fibres with small diameter and possessing high activity of oxidating enzymes. According to these signs, esophageal myons can be ascribed to the first type of muscular fibres. Simultaneously, a high activity of myofibrillar ATPh is revealed in them. Thus, according to histochemical indices, cross-striated esophageal muscular fibres of mammals are quick and slowly fatiqued fibres.


[Differentiation of muscle fibers of various types during postnatal development of the ventral serrate muscle in a rat]. In the denticulate ventral muscle of Wistar rats at the age of 1 day--2 months activity of NAD-N-dehydrogenase, succynic dehydrogenase and cytochrome oxidase has been determined in transversal cryostat sections. Quantitative estimation of the enzymes activity has been carried on by the plag-method. With age, general tendency to increasing activity of the enzymes mentioned is noted, but the dynamics of the increase is peculiar for every enzyme. Analysing the histograms on muscular fibre distribution according to their optical density, it is possible to estimate the dynamics. Simultaneously, the width of variational series, central statistical moments, indices of asymmetry and excess are also estimated. The whole course of the muscular fibre development, in accordance to the range and moments of distribution, can be devided into four main stages: stable, initial stage, slow increase of events, rapid increase of events and stabilization of the process. The stages mentioned pass gradually one into another making it possible to mark transitional stages (5--11, 15--19, 34--60 days). Using standard indices, it is possible to characterize more strictly the processes occurring in the course of muscular fibre differentiation. Lack of parallelism in the dynamics of asimilarity and excess can be treated as variety in differentiation of muscular fibres with middle and large optic density, and parallelism in dynamics--as their simultaneous differentiation. By comparing the curves it can be concluded that up to the 12--14th days, variety in differentiation of muscular fibres occurs, while after the 14th day their differentiation is more regular and simultaneous. The method of cytophotometry with subsequent mathematical processing of the results helps to determine the stages of muscular fibres differentiation.


The resistance welded stainless steel interface. The results obtained using single impulse and multiple impulse welds for joining orthodontic band and wire material showed that the single impulse produced an autogenous nugget of fused parent material and several nuggets were formed with the multiple impulse weld. Excessive current was deleterious.


Prescribing in Australia. Part 2.

Prescribing in Australia. Part 2. A brief history of the Australian Pharmaceutical Benefits Scheme is given. Available statistics in recent years have been analyzed indicating the more frequently prescribed drugs both individually and in the major pharmacological groups. Part 1, which covers antibiotics and analgesics was published in AFP, February, 1979.


Urinary lysosomal hydrolases in mucolipidosis II and mucolipidosis III. Investigation of the binding characteristics of acid beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase, alpha-D-galactosidase and alpha-L-fucosidase from patients with mucolipidosis II and mucolipidosis III to concanavalin A--Sepharose 4B revealed a 2--10-fold decrease in the proportion of enzyme activities from patients with mucolipidoses II and III that adsorbed on the lectin. Neuraminidase treatment of the unadsorbed enzyme fraction did not significantly increased the proportion of enzyme activities that bound to the concanavalin A--Sepharose 4B. Characterization of acid beta-D-galactosidase from the adsorbed and unadsorbed enzyme fractions of mucolipidosis II and mucolipidosis III patients demonstrated identical apparent Km values of 0.22 mM with respect to 4-methylumbelliferyl beta-D-galactopyranoside, altered pH--activity profiles and heterogeneous isoelectric-focusing patterns. The results of this study support the suggestion of an alteration of a post-translational modification (possibly glycosylation) occurring in mucolipidosis II and mucolipidosis III common to the lysosomal hydrolases that affects the mannoserelated properties of these enzymes.


The adsorption of proteins and protein--dodecyl sulphate complexes on N-(3-carboxypropionyl)aminodecyl-Sepharose. Equilibrium and kinetic aspects of the binding of several proteins to N-(3-carboxypropionyl)aminodecyl-Sepharose, an amphiphilic ampholytic adsorbent, were studied at 22 degrees C, pH 7.0, I 0.10--0.12. In the absence of detergents Scatchard plots are linear for human haemoglobin and soya-bean trypsin inhibitor, but non-linear for bovine serum albumin, which is also adsorbed more tightly than the other two proteins. The introduction [corrected] of 3.5mM-sodium dodecyl sulphate causes dramatic increases in the amounts and affinities of serum albumin and haemoglobin adsorbed, but has relatively little effect on the trypsin inhibitor. At concentrations of sodium dodecyl sulphate greater than about 10mM there is a fall in the binding of all proteins, owing to competition from the detergent for binding sites on the adsorbent, and a tendency towards more uniform behaviour by different proteins. Kinetic experiments suggest that in the absence of the detergent haemoglobin and serum albumin are adsorbed initially by mainly ionic forces, but that subsequently hydrophobic forces become dominant. Addition of 3.5 mM-sodium dodecyl sulphate causes pronounced changes in the time course of adsorption of haemoglobin and serum albumin, the nature of the changes being different for each protein. The significance of these results is discussed.


Circular-dichroism studies of ligand binding to dihydrofolate reductase from Lactobacillus casei MTX/R. Circular-dichroism spectra (200--450 nm) were recorded for Lactobacillus casei MTX/R dihydrofolate reductase and its complexes with substrates, inhibitors and coenzymes. These spectra are compared with those reported by others for dihydrofolate reductase from other sources. The binding of NADP+ or NADPH is associated with the perturbation of one or more aromatic amino acid residues, and there is marked enhancement of the negative c.d. band at 340 nm arising from the dihydronicotinamide chromophore of NADPH. The substrates folate and dihydrofolate give rise to substantial extrinsic c.d. bands on binding, which show a number of specific differences between enzymes from different sources. The binary complexes between the enzyme and the inhibitors methotrexate or trimethoprim also show strong c.d. bands, and these are qualitatively very similar for all dihydrofolate reductases studied so far. The ternary complexes between enzyme, NADPH and trimethoprim or methotrexate are very different from the sum of the spectra of the binary complexes. Trimethoprim leads to the disappearance of the 340 nm c.d. band of bound NADPH, whereas in the methotrexate--NADPH--enzyme ternary complex a "couplet" c.d. spectrum is observed at long wavelengths. Analysis of this latter feature suggests that it arises from a direct interaction between the dihydronicotinamide and pteridine rings in the ternary complex.


Kinetic studies of dogfish liver glutamate dehydrogenase. Initial-rate studies were made of the oxidation of L-glutamate by NAD+ and NADP+ catalysed by highly purified preparations of dogfish liver glutamate dehydrogenase. With NAD+ as coenzyme the kinetics show the same features of coenzyme activation as seen with the bovine liver enzyme [Engel & Dalziel (1969) Biochem. J. 115, 621--631]. With NADP+ as coenzyme, initial rates are much slower than with NAD+, and Lineweaver--Burk plots are linear over extended ranges of substrate and coenzyme concentration. Stopped-flow studies with NADP+ as coenzyme give no evidence for the accumulation of significant concentrations of NADPH-containing complexes with the enzyme in the steady state. Protection studies against inactivation by pyridoxal 5'-phosphate indicate that NAD+ and NADP+ give the same degree of protection in the presence of sodium glutarate. The results are used to deduce information about the mechanism of glutamate oxidation by the enzyme. Initial-rate studies of the reductive amination of 2-oxoglutarate by NADH and NADPH catalysed by dogfish liver glutamate dehydrogenase showed that the kinetic features of the reaction are very similar with both coenzymes, but reactions with NADH are much faster. The data show that a number of possible mechanisms for the reaction may be discarded, including the compulsory mechanism (previously proposed for the enzyme) in which the sequence of binding is NAD(P)H, NH4+ and 2-oxoglutarate. The kinetic data suggest either a rapid-equilibrium random mechanism or the compulsory mechanism with the binding sequence NH4+, NAD(P)H, 2-oxoglutarate. However, binding studies and protection studies indicate that coenzyme and 2-oxoglutarate do bind to the free enzyme.


Zinc(II) binding to apo-(bovine erythrocyte superoxide dismutase). The binding of zinc(II) ions to apo-(bovine erythrocytes superoxide dismutase) was studied by 1H n.m.r. spectroscopy. Two zinc(II) ions bind to each subunit of the apoenzyme, and the first has a binding constant at least an order of magnitude larger than the second. The nature of the spectral changes that occur on binding the first zinc(II) ion are interpreted in terms of a change in the structure of the protein around the active site to one very similar to that of the holoenzyme, thus pre-forming the second zinc(II) binding site. The binding of the second zinc(II) ion effects changes in the environment of only those residues involved in its co-ordination.


Enzyme activity in partly dissociated fragments of rat intestinal maltase/glucoamylase. Pure rat intestinal maltase/glucoamylase was partially inactivated in 1% sodium dodecyl sulphage by heating at 40--70 degree C for 5 min at pH 7.5, or by lowering the pH to 5.4--6.6 at 24 degree C. When partially active preparations were electrophoresed in the presence of sodium dodecyl sulphate, a complicated protein band pattern of incompletely dissociated fragments of the enzyme was observed. Complete dissociation of the enzyme in sodium dodecyl sulphate, induced by boiling or by pH values below 5.4, was accompanied by total loss of enzyme activity and simplification of the protein pattern to five major species. Although the original enzyme band was absent from some partially dissociated preparations, enzyme activity was present and was associated with several transient protein bands on the gels. Maltase and alpha-glucosidase activities were detected in these bands, but glucoamylase activity was absent.


Some kinetic and other properties of the isoenzymes of aspartate aminotransferase isolated from sheep liver. A method for the purification of mitochondrial isoenzyme of sheep liver aspartate aminotransferase (EC 2.6.1.1) is described. The final preparation is homogeneous by ultracentrifuge analyses and polyacrylamide-gel electrophoresis and has a high specific activity (182 units/mg). The molecular weight determined by sedimentation equilibrium is 87,100 +/- 680. The amino acid composition is presented; it is similar to that of other mitochondrial isoenzymes, but with a higher content of tyrosine and threonine. Subforms have been detected. On isoelectric focusing a broad band was obtained, with pI 9.14. The properties of the mitochondrial aspartate aminotransferase are compared with those of the cytoplasmic isoenzyme. The Km for L-aspartate and 2-oxoglutarate for the cytoplasmic enzyme were 2.96 +/- 0.20 mM and 0.093 +/- 0.010 mM respectively; the corresponding values for the mitochondrial form were 0.40 +/- 0.12 mM and 0.98 +/- 0.14 mM. Cytoplasmic aspartate aminotransferase showed substrate inhibition by concentrations of 2-oxoglutarate above 0.25 mM in the presence of aspartate up to 2mM. The mitochondrial isoenzyme was not inhibited in this way. Pi at pH 7.4 inhibited cytoplasmic holoenzyme activity by up to about 60% and mitochondrial holoenzyme activity up to 40%. The apparent dissociation constants for pyridoxal 5'-phosphate were 0.23 micrometer (cytoplasmic) and 0.062 micrometer (mitochondrial) and for pyridoxamine 5'-phosphate they were 70 micrometer (cytoplasmic) and 40 micrometer (mitochondrial). Pi competitively inhibited coenzyme binding to the apoenzymes; the inhibition constants at 37 degree C were 32 micrometer for the cytoplasmic isoenzyme and 19.5 micrometer for the mitochondrial form.


Kinetics and mechanism of action of aldehyde reductase from pig kidney. An improved procedure for purifying aldehyde reductase is described. Utilization of Blue Dextran--Sepharose 4B and elimination of hydroxyapatite chromatography greatly improves the yield and ease of purification. Starting with 340 g of kidney tissue (two pig kidneys) approx. 50 mg of purified reductase may be routinely and reproducibly obtained. The purified reductase was used to establish the kinetic reaction mechanism of the enzyme. Initial-velocity analysis and product-inhibition data revealed that pig kidney aldehyde reductase follows an Ordered Bi Bi reaction mechanism in which NADPH binds first before D-glyceraldehyde. The limiting Michaelis constants for D-glyceraldehyde and NADPH were 4.8 +/- 0.7 mM and  9.1 +/- 2.1 micrometer respectively. The mechanism is similar to that of another monomeric oxidoreductase, octopine dehydrogenase, towards which aldehyde reductase exhibits several similarities, but differs from that of other aldehyde reductases. Phenobarbital is a potent inhibitor of aldehyde reductase, inhibiting both substrate and cofactor non-competitively (Ki = 80.4 +/- 10.5 micrometer and 66.9 +/- 1.6 micrometer respectively). Barbiturate inhibition seems to be a common property of NADPH-dependent aldehyde reductases.


The mechanism of reduction of single-site redox proteins by ascorbic acid. The reduction of single-site haem and copper redox proteins by ascorbic acid was studied as a function of pH. Evidence is presented that indicates that the double-deprotonated ascorbate anion, ascorbate2-, is the reducing agent, and the pH-independent second-order rate constants for reduction by this species are given. Investigation of the temperature dependences of these rate constants have yielded the values of the activation parameters (delta H++ and delta S++) for reduction. These values, together with ligand-replacement studies, suggest that ascorbate2- acts as an outer-sphere reductant for these proteins. Reasons to account for the apparent inability of ascorbic acid to reduce the alkaline conformer of mammalian ferricytochrome c are suggested.


Artificially induced active transport of amino acid driven by the efflux of a sugar via a heterologous transport system in de-energized Escherichia coli. Consistent with the model of an H+ cotransport, amino acid uptake can be driven by a proton gradient generated by an efflux of sugar when the normal energy sources are suppressed. Heterologous countertransport is completely inhibited by uncouplers unlike homologous countertransport. Positive coupling was obtained with methyl thiogalactoside/proline, methyl thiogalactoside/phenylalanine, gluconate/proline; however, the poor coupling efficiency suggests a more complex sequence of reactions.


Transmembrane electrical potential and transmembrane pH gradient in the acidophile Thiobacillus ferro-oxidans. Thiobacillus ferro-oxidans is capable of using the oxidation of Fe2+ by O2 at pH 2.0 as the sole source of energy for growth and CO2 fixation. The bacterium maintains an intracellular pH of 6.5 over a range of external pH from 1.0 to 8.0, as measured by [14C]acetate and [3H]methylamine distribution. The membrane potential was estimated by the distribution of the lipid-soluble cation dibenzyldimethylammonium and the anion SCN-. At pH 2.0 (the pH of growth) during Fe2+ oxidation the transmembrane pH gradient is 4.5 units with an opposing membrane potential of -10mV, giving a proton electrochemical gradient of +256mV. This gradient is actively maintained.


[The influence of 2-ethyl-3-(4-hydroxy-benzoyl)- benzofuran on the contraction of pial arterial muscles (author's transl)]. By means of perivascular perfusion of thin sections of pial arteries with differently composed cerebrospinal fluid (CSF) the vessel diameters can be influenced. In cats, anesthetized with pentobarbital, addition of serotonin, norepinephrine or calcium to an artificial CSF which was infused through micropipettes into the perivascular space of the pial arteries, caused constrictions of the arteries contacting the modified CSF. Hyperventilation or perivascular perfusion of alkaline CSF also caused reduction of the pial arterial diameters. 2-Ethyl-3-(-hydroxy-benzoyl)-benzofuran (benzarone) inhibited or abolished the constrictions elicited by the above mentioned stimuli. The effect was dose-dependent. High doses of benzarone caused pial arterial dilations: The inhibition of the calcium induced constriction especially, seems worth mentioning since this ion plays a central role in the contraction mechanisms of the vascular smooth muscle cells.


The interaction of central acting drugs with glutamate dehydrogenase. The neuroleptics chlorpromazine, fluphenazine, triflupromazine and thioridazine seem to effect the aggregation of glutamate dehydrogenase by binding to one single site of the polypeptide chain, possibly the GTP site. This interaction could biologically be of importance for the degree of activity at high enzyme concentrations found in vivo. Amitriptyline and fluorazepam also bind to a specific single site of the polypeptide chain without effecting the aggregation of the oligomers. Because of the rather low affinity of these sites the binding of these drugs in vivo does not seem to cause a concentrating effect in the mitochondria.


[Studies on the biotransformation of reproterol. Structure determination of the main metabolite (author's transl)]. Biotransformation of 7-(3-[2-(3,5-dihydroxyphenyl)-2-hydroxy-ethylamino]-propyl)-theophylline (reproterol, Bronchospasmin), a beta 2-adrenergic drug recently introduced into therapeutic use, leads to the same main metabolite in animals and in man. By mass-spectroscopy and by synthesis its structure was shown to be tetrahydroisoquinoline derivative which is produced from reproterol by uptake of an additional carbon atom concomitant with cyclization.


Pharmacological studies on CS-430, a new psychotropic agent. 10-Bromo-11b-(2-fluorophenyl)-2,3,7,11b-tetrahydrooxazolo[3,2-d][1,4]benzodiazepin-6(5H)-one (CS-430) is a new psychotropic drug and has the following properties. The compound showed sleep-inducing effect at both 0.2 and 5 mg/kg (p.o.) in cynomolgus monkeys or at 3 mg/kg (p.o.) in rats. CS-430 showed selective conflict attenuating in rats or anticonvulsant effects in mice. Furthermore, CS-430 blocked non-discriminated (Sidman) avoidance response without severe impairment of the motor function but not discriminated (shuttle-box) avoidance response in rats. CS-430 also blocked selectively electroshock-induced fighting behavior of mice but not isolation-induced fighting behavior of mice and muricidal behavior induced by ablation of the olfactory bulbs. In addition, CS-430 produced specific potentiation of chlorprothixene-induced sleep and particular potentiation of thiopental sleep in mice. These effects of CS-430 were similar to those of nitrazepam, but differed from those of phenobarbital. The effect of CS-430 on the motor function was roughly 1/2 as potent as that of nitrazepam in the various tests.


Inhibitory effect of beta-adrenergic stimulants on the histamine reaction of the human skin. II. Studies with several derivatives in regard to the dosage used and reaction depending on time. In 56 volunteers the inhibitory effects of the beta-adrenergic stimulants fenoterol, salbutamol, isoproterenol, orciprenaline, terbutaline, epinephrine and phenylephrine were studied. There was a difference between the substances in regard to the applied dose and duration of inhibitory effect. The latter parameter particularly depended on the hydrosolubility of the individual substances. Fenoterol and salbutamol had a very low solubility and a very strong and long-lasting effect. The possibly stronger adhesive power of these drugs in the tissue according to their poor solubility and the site of action in the reactive systems of the skin are discussed giving preference to the vascular system over the mast-cells and cAMP.


Gaschromatography of clemastine. A study of plasma kinetics and biological effect. A method for estimation of clemastine in human plasma has been developed. By chronic acid oxidation the drug is degradated to chlorobenzophenone, which can then be analysed by gas-liquid-chromatography using electron-capture detection. The plasma levels of clemastine after oral and i.v. administration have been studied. Using different methods for extraction of the drug from plasma a metabolic degradation of the pyrrolidine part of clemastine is demonstrated. The metabolite proposed may be biologically active. The plasma level of the drug was found to be directly related to its biologic effect, measured as inhibition of the histamine-induced flare in human skin.


Kinetics of degradation of cefazolin and cephalexin in aqueous solution. The kinetics of degradation of cefazolin and cephalexin in aqueous solution were investigated at 60 degrees C and constant ionic strength over the entire pH range. The observed degradation rates were obtained by measuring the residual cephalosporin and were shown to follow pseudo-first-order-kinetics. They were influenced significantly by solvolytic and hydroxide ion catalysis. No primary salt effect was observed in the acid or basic pH region. Of the buffer systems employed in the kinetics studies only the phosphate buffer system showed a catalytic effect. The pH-rate profile for cefazolin showed a degradation minimum between pH 5.5 and 6.5. Cephalexin did not show a pH minimum in that region. The apparent energies of activation were determined for cefazolin and cephalexin at pH 5.5 and were calculated to be 24.3 Kcal/mole and 26.2 Kcal/mole, respectively. The agreement between the calculated theoretical pH-rate profiles and the experimental points for both compounds support the hypothesis presented concerning the reactions involved in their respective degradation pathways.


[Effect of cyclophosphamide and some derivatives on the cortisone induction of tyrosine-aminotransferase in rat liver (author's transl)]. The enzyme tyrosine-aminotransferase is induced in the liver of adrenalectomised rats by cortisone acetate with doses higher than 5 mg/kg. This induction is increased by the application of cyclophosphamide. In the case of 4-hydroperoxy-cyclophosphamide (active principle of Endoxan) the induction is considerably higher,  whereas the immunosuppressive substance sufosamide (Asta 5122) is less effective. With the compound Asta 4968 which lacks carcinostatic and immunosuppressive effects no stimulation can be observed.


[Preparation of iduronate sulfatase from the human placenta]. The preparation of the enzyme iduronate sulfatase from human placenta has been undertaken. The substrate O-(alpha-L-idopyranosyluronic acid 2-sulfate) (1 leads to 4)-2,5-anhydro-D-[3H]mannitol 6-sulfate was used to measure the enzymatic activity. The enzyme shows a pH optimum of 4.0 in 0.1 M sodium formiate or acetate buffer. Chromatography on DE-52 gives a 5.4 fold purification. The enzyme is inhibited by NaCl or KCl: in 20 mM salt the reaction rate was only 63% and 34% respectively. Inhibition by salt can be removed by extensive dialysis after the chromatographic step.


A clinical view of clobazam.

A clinical view of clobazam. 1 The results so far published in the literature, together with our own results, show that in clobazam (Frisium) we have a daytime tranquilizer which is comparable to the newest benzodiazepine derivatives. 2 It is particularly indicated in disorders of the sleeping-waking rhythm, and sometimes also in psychovegetative disorders associated with epilepsy. 3. Its tranquillizing profile of action, the low incidence of side-effects and the absence of interactions with other drugs mean that its applications are varied, not only in neuropsychiatry but also in other medical disciplines. 4 The fact that therapeutic doses do not seem to impair driving ability to render patients incapable of carrying on their profession, means that it is suitable for the treatment of outpatients.


Evaluation of the effects of clobazam, A 1,5 benzodiazepine, on mood and psychomotor performance in clinically anxious patients in general practice. 1 The methodology of clinical trails of anxiolytic drugs carried out in general practice conditions is discussed. Particular problems include: the selection of suitable patients; placebo effects; the influence of non-specific variables such as life-events; and patient compliance. 2 Clobazam, a novel 1,5 benzodiazepine, and diazepam were compared with placebo in a double-blind group comparative trial in general practice, which was designed to avoid as many confounding variables as possible. Anxiety-reducing effects were evaluated and at the same time the effects of the drugs on psychomotor performance were examined. 3 Both clobazam and diazepam produced significant improvements in anxiety ratings on the Hamilton Anxiety Scale and the Morbid Anxiety Inventory, whereas placebo did not. 4 The placebo group demonstrated a significant improvement in performance on a pursuit rotor and digit symbol substitution test (DSST), whereas the diazepam group's performance did not change. The clobazam group showed improvement in both tests, significantly so in the DSST. This suggests that diazepam produces impairment in performance, which had negated the practice effects seen in the placebo group, whereas clobazam did not seem to produce similar impairment.


Single daily dose treatment of anxiety with clobazam or dipotassium clorazepate. 1 Forty-four clinically anxious patients entered a comparative double-blind trial of clobazam 20 mg, clobazam 30 mg and dipotassium clorazepate 15 mg, all drugs given as a single dose at night. 2 Assessment by the Hamilton Anxiety Scale, Morbid Anxiety Inventory (Salkind) and a Visual Analogue Scale showed a statistically significant improvement for all treatment groups after 2 weeks, with continued improvement after a further 2 weeks. 3 Daytime drowsiness was the commonest side-effect in all treatment groups but there was a tendency for a lower incidence in patients on clobazam. There was no evidence of a dose-related incidence of drowsiness in the clobazam 20 mg and 30 mg groups. Other side-effects were few and nonspecific. 4 Clobazam is a 1,5-benzodiazepine with an elimination half-life of 18 hours. When given in single doses of 20-30 mg at night it has an equivalent effect to dipotassium clorazepate 15 mg.


Clobazam: uncontrolled and standard controlled clinical trials. 1 In an uncontrolled clinical trial, carried out in 11 psychiatric patients with the clinical diagnoses of anxiety neurosis and depressive neurosis, clobazam, a new benzodiazepine preparation, in the dosage range 10-60 mg daily produced statistically significant improvement in the total and both factor scores of the Hamilton Anxiety Scale (HAM-A). The lowest mean total HAM-A scores occurred with a mean clobazam dosage of 48 mg daily. 2 Results of the uncontrolled clinical trial were further substantiated in a standard-controlled clinical study in which no statistically significant difference between the therapeutic effectiveness of clobazam and diazepam could be revealed. The lowest mean total HAM-A scores occurred with a mean clobazam dosage of 49 mg daily. There was a lower incidence of adverse effects reported in patients receiving clobazam than in those taking the control drug (diazepam).


Review of clinical studies on clobazam.

Review of clinical studies on clobazam. 1 Clobazam was compared with placebo and diazepam in 45 double-blind studies. 2 Clobazam dosage ranged from 5 mg daily (paediatric patients) to 120 mg daily (psychiatric in-patients). Usual daily dosage in out-patient therapy ranged from 20-30 mg clobazam. 3 Treatment duration varied from a few days up to 3.5 years. The usual duration of treatment was 2-4 weeks. 4 Clobazam was shown to be an efficacious and well tolerated anxiolytic agent in various neurotic and psychosomatic disorders. 5 Dosages showing distinct anxiolytic effects in out-patient therapy did not impair psychomotor performance.


Clobazam: pharmacological and therapeutic profile. 1 Clobazam is a 1,5-benzodiazepine the pharmacological profile of which differs from that of the 1,4 benzodiazepines in that it displays a wide separation of psychosedative or 'tranquillizing' properties from impairment of motor coordination. This is associated with a relative lack of muscle relaxant activity. 2 In man it seems to be an effective anxiolytic agent when given in daily doses of 20-30 mg but produces minimal effects on psychomotor performance at these dose levels. 3 Clobazam has a long elimination half-life of about 18 h; the main metabolite is N-desmethylclobazam. Clobazam seems to share the good safety and tolerability characteristics of the 1,4 benzodiazepines.


Clobazam: chemical aspects of the 1,4 and 1,5-benzodiazepines. 1 The structures and chemistry of the 1,4- and 1,5-benzodiazepines are compared. These two groups of drugs differ in respect to basicity, lipophilicity and electronic charge distribution. 2 The 1,4-benzodiazepine diazepam has a weakly basic imine group in contrast to the acid methylene protons of clobazam. 3 The imine function of diazepam is more lipophilic than the carboxamide group of clobazam with its rather hydrophilic character. 4 Diazepam and clobazam differ in electronic charge distribution and hence in the location of the chemically reactive centres, the imine partial structure in diazepam and the malonic acid portion of clobazam.


In vitro studies on GABA release.

In vitro studies on GABA release. 1 Recent studies have demonstrated growing evidence for a primary action of the benzodiazepines on gabaminergic neurones which induces a facilitation of gamma-aminobutyric acid (GABA)-mediated neurotransmission. As enhancement of GABA release has been suggested to account for their activation of GABA mechanisms, the effect of diazepam and clobazam, and of several other psychotropic drugs, on stimulated GABA release have been studied. 2 Using rat brain cortex slices saturated with [3H]-GABA, the electrically stimulated overflow of GABA is reduced in a concentration-dependent manner in the presence of both diazepam and clobazam. 3 The benzodiazepine-induced reduction in GABA overflow during electrical stimulation is antagonized by the GABA receptor blocker bicuculline, whereas bicuculline alone at 10(-6) M concentration does not change the overflow. 4 Among some other centrally active drugs tested, hexobarbitone and the 'second messenger; cyclic GMP also induce a significant but less marked reduction in GABA release. 5 A schematic model of a central gabaminergic synapse is proposed, which may explain the benzodiazepine effects on stimulated GABA release by suggesting an inhibitory feedback control of transmitter release mediated by presynaptic GABA receptors ('autoreceptors').


Toxicology of clobazam.

Toxicology of clobazam. 1 Extensive toxicological investigations of clobazam are summarized. 2 LD50 values after oral administration ranged from 100 mg/kg in the dog to 6000 mg/kg in the rat. 3 Long-term repeated dose studies have shown withdrawal effects in the dog and monkey (notably convulsions leading to death), similar to those known to occur with other benzodiazepines. 4 It is concluded that the overall safety and tolerability of clobazam have been demonstrated in the preclinical animal studies.


Assay of clobazam in human serum by a radioreceptor technique. 1 Clobazam has been assayed in human serum using a radioreceptor technique. 2 The assay is simple and sufficiently sensitive for detecting serum levels up to 48 h after oral administration of a usual clinical dose. It has the advantage of measuring the parent compound plus active metabolites. 3 Serum clearances and elimination half-lives have been determined for five subjects.


Pharmacokinetics of single and multiple doses of clobazam in humans. 1. The pharmacokinetics of clobazam and its biotransformation product N-desmethylclobazam were investigated after single and multiple doses in normal subjects. 2. The relevant physicochemical properties of clobazam were measured and are presented. Different assay methods (radiochemical, fluorimetric and gas chromatographic) were applied and the results correlated. 3. After single doses the pharmacokinetic profile of clobazam includes time to peak levels 1--4 h after dosing, peak levels increasing linearly with the logarithm of dose, and terminal half-lives of about 18 hours. At least 87% of an oral dose is absorbed, as indicated by urinary recovery of labelled material. 4. In multiple-dose studies unchanged clobazam levelled off at minimum steady-state concentrations within one week of dosing. During 28 d of medication N-desmethylclobazam accumulated to near steady-state levels about eight times higher than those of the unchanged compound. 5. No pharmacokinetic interactions were discovered between clobazam and the antidepressant nomifensine.


Effects of benzodiazepines on psychomotor performance. 1 The literature relating to the effects of benzodiazepines on psychomotor performance is critically reviewed. 2 The multiple and diverse psychomotor tests used are assessed according to their ability to demonstrate differences between drugs. 3 Three general conclusions are: (1) The speed with which simple acts of a repetitive nature are performed may be impaired by benzodiazepines. (2) learning and immediate memory will also be impaired. (3) there is relatively little indication that well established higher mental faculties are adversely involved.


Psychomotor changes during initial day of benzodiazepine medication. 1 The detracting psychomotor effects of diazepam (5 mg three times daily) and clobazam (an investigational 1,5-benzodiazepine) were compared with placebo effects over the course of the initial day of medication. Tests were administered at hourly intervals and the data were analyzed from the standpoint of contrasts at each session and from the standpoint of trends that accrued during the course of the day. 2 It is concluded that among normal volunteers diazepam 5 mg three times daily may be near the threshold for detracting psychomotor consequences during the initial day and that clobazam seems to be without detracting consequences and may have some enhancing effects.


Pharmacology of anti-anxiety drugs with special reference to clobazam. 1 In several studies clobazam exhibited a potency which ranges between that of chlordiazepoxide and diazepam. Its anxiolytic and anti-aggression effects are produced by doses usually ranging below those that cause disorders in motor activity. 2 This separation was demonstrable to an even greater degree with the desmethyl metabolite. The activity of the metabolite, however, was weaker than that of the original substance. 3 The advantage of clobazam compared with the 1.4 benzodiazepines lies mainly in the fact that motor activity is influenced only after very high doses, these doses being markedly above those required to induce tranquillizing and anti-agression activities. 4 Clobazam has no marked effect on the cardiovascular system, respiration of excretion.


Some aspects of the effects of clobazam on human psychomotor performance. 1 Three studies are described, the first being a comparison of the effects of acute night-time doses of clobazam 20 mg, amylobarbitone sodium 100 mg, nitrazepam 5 mg and placebo, on choice reaction time, critical flicker fusion (CFF) and stabilometer performance. Clobazam improved early morning performance on a choice reaction test, in contrast to the other two active drugs. 2 Repeated doses of clobazam 10 mg three times daily, chlordiazepoxide 10 mg three times daily and diazepam 5 mg three times daily were given for 5 days. Again clobazam did not produce any impairment of psychomotor performance, and noticeably increased CFF thresholds. 3 The effects of an acute night-time dose of clobazam 20 mg on psychomotor performance the morning after night-time medication were correlated with the neuroticism scores (on the EPI) of the subjects. Clobazam exerts a differential effect on psychomotor performance dependent on the basic personality trait. 4 Clobazam seems to differ significantly from the 1,4-benzodiazepines in that, although it reduces anxiety, it does so without any apparent impairment of psychomotor performance.


Differential effects of the 1,4 and 1,5 benzodiazepines on performance in healthy man. 1 The immediate and residual effects on performance of benzodiazepines differ, and the differences are important in the use of these drugs. 2. Diazepam and its hydroxylated metabolites, temazepam and oxazepam, each possess hypnotic activity, and have effects on performance limited to the sleep period. The demethylated metabolite of diazepam, nordiazepam, and its precursor, potassium clorazepate, which also possess hypnotic activity, have a longer duration of action, although the next day anxiolytic effect is accompanied by only minimal effects on performance. The 1.5 benzodiazepine, clobazam, seems to have minimal immediate effects on performance. 3 Diazepam and its hydroxylated metabolites, temazepam and oxazepam, would be useful in the management of insomnia without psychopathology in those cases in which residual effects on performance must be avoided. Nordiazepam and potassium clorazepate would be appropriate for insomnia secondary to day-time anxiety, and clobazam may be useful as a day-time anxiolytic. 4 It is emphasized that more work needs to be carried out on the effects of anxiolytics on performance before one can be certain that ingestion during the day would be without any deleterious effects on skilled work.


Studies of clobazam and car-driving.

Studies of clobazam and car-driving. 1 The methodology used in studies to assess the effects of drugs on car-driving performance is reviewed. 2 Clobazam 20 mg, diazepam 10 mg or placebo were administered daily for 3 d to 24-male students with a high neuroticism score (on the Cattell Personality Factors Questionnaire). 3 Car driving performance was assessed on the second day in real traffic conditions; tests of attention and concentration and subjective assessments were made on the third day. 4 Diazepam 10 mg significantly impaired braking reaction time in comparison with clobazam 20 mg and placebo (P less than 0.01). Subjects also reported feeling more 'depressed' and lethargic after diazepam.


Electrostatic effects in hemoglobin: Bohr effect and ionic strength dependence of individual groups. The electrostatic treatment applied in the preceding paper in this issue [Matthew, J. B., Hanania, G.I.H., & Gurd, F.R.N. (1979) Biochemistry (preceding paper in this issue)] to the titration behavior of individual groups in human deoxyhemoglobin and oxyhemoglobin was applied to the computation of the alkaline Bohr effect at various values of ionic strength. The enhanced proton binding of deoxyhemoglobin in the pH range of 6--9 was accounted for at ionic strength 0.01 M by the effects of the unique charge distributions of ionizable groups in the two quaternary states. At ionic strength 0.10 M the effects of 2--4 bound anions had to be considered in addition in the deoxyhemoglobin charge configuration. At the higher ionic strength 10 groups per tetramer contributed to the Bohr effect, whereas 28 groups were contributory at the lower ionic strength. The ionic strength dependence of individual groups in the two tetrameric structures as well as in the alpha-chain monomer was explained in terms of the electrostatic treatment. This examination showed that the differences in electrostatic behavior of deoxy- and oxyhemoglobin follow from particular dissymmetries in their configurations with respect to charge and static solvent accessibility.


s-Cis and s-trans isomerism of the His-Pro peptide bond in angiotensin and thyroliberin analogues. The dipeptide His-Pro isomerizes from all-s-trans to partly s-cis when titrated in D2O from acidic to neutral pD as observed by 13C and 1H nuclear magnetic resonance of the proline side chain. This isomerization is reported by the His C-2 and C-4 protons and carbons which show distinct, well-resolved resonances for each isomer. The influence of the His-Pro peptide bond rotational state on the histidine protons far removed from the bond has not been previously observed in model compounds or peptides. The peptides thyroliberin (TRH), [3-MeHis2]-TRH, and [3-MeHis6]-, [Sar1,Al8]-, and Nalpha-acetylangiotensin II were found to similarly isomerize from all-s-trans to partly s-cis as reported by their His C-2 and C-4 proton resonances. The His C-2 and C-4 protons in the peptides [1,3-diMeHis2]-TRH and [1-MeHis6]-, and [homoHis6]-angiotensin do not report this isomerization. Angiotensin II has previously been found to exhibit the same isomerization. The reporting of the s-trans to s-cis isomerization by the His C-2 proton appears to be correlated with the known potencies of the five angiotensin peptides in rat uterine strips and of the three TRH peptides by radioimmunoassay of released thyrotropin.


Relaxation studies of enzymes: concentration and pH dependence of isomerization phenomena. Equations have been developed for the relaxation times for a variety of mechanisms involving enzyme isomerization coupled to proton transfers. The concentration and pH dependences of the relaxation time have been calculated and graphed for a number of representative mechanisms. We find that for most of the mechanisms examined, the relaxation time is not only pH but also strongly concentration dependent. The concentration dependence results from finite perturbations of the hydrogen ion concentration. For the systems tested, the relaxation time shows a clear concentration dependence at enzyme concentrations below 200 microM.


Relaxation studies of enzymes: rapid isomerization in deoxyribonuclease I. Temperature-jump relaxation studies in deoxy-ribonuclease I were carried out at 10 degrees C and [I] = 0.1 M. The single observed relaxation time, which varied from 10(-4) to 10(-5) s, was characterized as a function of enzyme concentration, pH, and indicator concentration. The concentration and pH dependences of the relaxation time are in quantitative agreement with a mechanism involving an isomerization of the enzyme coupled to a rapid proton ionization process. The best fit forward and reverse isomerization rate constants are 6.5 X 10(3) and 7.2 X 10(4) s-1, respectively; the apparent pK is 5.7. The addition of urea brought about reductions in both the amplitude of the relaxation effect and the enzyme activity.


Conformational transitions and vibronic couplings in acid ferricytochrome c: a resonance Raman study. Resonance Raman spectral changes in ferricytochrome c as a function of pH between 6.7 and 1.0 are reported and the structural implication is discussed in terms of the "core-expansion" model advanced by L. D. Spaulding et al. [(1975) J. Am. Chem. Soc. 97, 2517]. The data are interpreted as indicating the iron in high-spin ferricytochrome c (at pH 2.0) with two water molecules as axial ligands lies in the plane of the porphyrin ring. At pH 1.0 there is a different high-spin form of cytochrome c which has an estimated iron out-of-plane distance of approximately 0.46 A. The effect of a monovalent anion at pH 2.0 is to produce a thermal spin mixture with predominant low-spin species. Excitation at approximately 620 nm in acid cytochrome c (pH 2.0) enhances only three depolarized ring vibrations at 1623, 1555, and 764 cm-1. Marked enhancement of depolarized modes relative to polarized and anomalously polarized modes is attributed to the vibronic coupling between porphyrin pi leads to pi and porphyrin pi leads to iron (dpi) charge-transfer states.


Specificity of guinea pig liver transglutaminase for amine substrates. The amine specificity of guinea pig liver transglutaminase, a model enzyme for endo-gamma-glutamine:epsilon-lysin transferases, was explored with the aid of synthetic substrates of high apparent affinities. As exemplified by dansyl- (5-dimethylamino-1-naphthalenesulfonyl), (2,4-dinitrobenzenesulfonyl)-, and (2,4,6-triisopropylbenzenesulfonyl)-cadaverines--each of which showed affinities of approximately 4 x 10(7) M-1--the best amine substrates carried a large hydrophobic substituent attached to an alkylamine side chain of about 7.2 A in length. Altogether, our results point to the importance of a hydrophobic binding region in the enzyme from where the alkyl side chain reaches into a narrow crevice toward the active center and positions the primary amine of the substrate for attacking the carbonyl group of the acyl enzyme intermediate.


A channel mechanism for electrogenic ion pumps. A model of active ion transport is analyzed in which an essential part of the pumps molecule is an ion channel. Ion translocation in the channel is described as a series of jumps between binding sites which are separated by energy barriers. Pumping action results from a transient energy-dependent modification of the barrier structure of the channel and requires only minor conformational changes of the pump molecule. This model is applied to the light-driven proton pump of Halobacterium and to redox-coupled proton pumps in the mitochondrial respiratory chain. Similar considerations may be used to describe ATP-dependent ion transport.


Coordination of Gly-Tyr . Pd(II) complex to ATP and ADP nucleotides. The coordination of the glycyl-L-tyrosinate . Pd(II) complex to adenosine 5' -diphosphate (ADP) and adenosine 5' -triphosphate (ATP) has been studied using 13C, 1H NMR and electronic spectral methods. Two dominant species have been found in solution, a monomeric ternary complex with Gly-Tyr . Pd(II) bound to the N-1 purine nitrogen and a dimer in which two dipeptide . Pd(II) complex molecules are coordinated to the nucleotide by N-1 and N-7 nitrogens. Monomeric ternary complexes having metal coordination to N-7 were not detected. The influence of the aromatic ring of tyrosine upon the chemical shifts for the bonded nucleotide molecule suggest that the plane of the purine ring and of the Gly-Tyr . Pd(II) complex are almost perpendicular to each other.


The enzymatic preparation of [alpha-(32)P]nucleoside triphosphates, cyclic [32P] AMP, and cyclic [32P] GMP. A method has been developed for the enzymatic preparation of alpha-(32)P-labeled ribo- and deoxyribonucleoside triphosphates, cyclic [(32)P]AMP, and cyclic [(32)P]GMP of high specific radioactivity and in high yield from (32)Pi. The method also enables the preparation of [gamma-(32)P]ATP, [gamma-(32)P]GTP, [gamma-(32)P]ITP, and [gamma-(32)P]-dATP of very high specific activity and in high yield. The preparation of the various [alpha-(32)P]nucleoside triphosphates relies on the phosphorylation of the respective 3'-nucleoside monophosphates with [gamma-(32)P]ATP by polynucleotide kinase and a subsequent nuclease reaction to form [5'-(32)P]nucleoside monophosphates. The [5'-(32)P]nucleoside monophosphates are then converted enzymatically to the respective triphosphates. All of the reactions leading to the formation of [alpha-(32)P]nucleoside triphosphates are carried out in the same reaction vessel, without intermediate purification steps, by the use of sequential reactions with the respective enzymes. Cyclic [(32)P]AMP and cyclic [(32)P]GMP are also prepared enzymatically from [alpha-(32)P]ATP or [alpha-(32)P]GTP by partially purified preparations of adenylate or guanylate cyclases. With the exception of the cyclases, all enzymes used are commerically available. The specific activity of (32)P-labeled ATP made by this method ranged from 200 to 1000 Ci/mmol for [alpha-(32)P]ATP and from 5800 to 6500 Ci/mmol for [gamma-(32)P]ATP. Minor modifications of the method should permit higher specific activities, especially for the [alpha-(32)P]nucleoside triphosphates. Methods for the use of the [alpha-(32)P]nucleoside phosphates are described for the study of adenylate and guanylate cyclases, cyclic AMP- and cyclic GMP phosphodiesterase, cyclic nucleotide binding proteins, and as precursors for the synthesis of other (32)P-labeled compounds of biological interest. Moreover, the [alpha-(32)P]nucleoside triphosphates prepared by this method should be very useful in studies on nucleic acid structure and metabolism and the [gamma-(32)P]nucleoside triphosphates should be useful in the study of phosphate transfer systems.


DNA-protein crosslinking by trans-platinum(II)diamminedichloride in mammalian cells, a new method of analysis. DNA-protien crosslinks produced in mouse leukemia L1210 cells by trans-Pt(II)diamminedichloride were quantitated using the technique of DNA alkaline elution. DNA single-strand segments that were or were not linked to protein were separable into distinct components by alkaline elution after exposure of the cells to 2--15 kR of X-ray. Protein-linked DNA strands were separated on the basis of their retention of filters at pH 12 while free DNA strands of the size generated by 2--15 kR of X-ray passed rapidly through the filters. The retention of protein-linked DNA strands was attributable to adsorption of protein to the filter under the conditions of alkaline elution. The results obeyed a simple quantitative model according to which the frequency of DNA-protein crosslinks could be calculated.


Effect of substrate properties on the activity of lysosomal cholesteryl ester hydrolase. The effects of the substrate properties on the catalytic activity of lysosomal cholesteryl ester hydrolase from rat liver have been examined with three standard substrate types: vesicle, micelle and emulsion. The pH optimum of the enzyme coincided to 4.5--5.0 with the substrate types employed. The apparent Km values were 15.3, 14.3 and 7.3 microM for vesicle, micelle and emulsion substrates, respectively. In the systems used in this study reaction products, cholesterol and oleic acid, and the nonionic surfactant Tween 80 and Triton X-100 Had an inhibitory effect. The emulsifier phosphatidylcholine and the charged phospholipid phosphatidic acid stimulated the activity. The mixed micelle of sodium taurocholate and phosphatidylcholine was the most potent substrate vehicle. With dipalmitoyl phosphatidylcholine vesicles the enzyme showed maximal activity at the gel-liquid-crystalline transition temperature of the phospholipid. The possible physiological significance of the lysosomal cholesteryl ester hydrolase is discussed with special reference to the form of the substrate.


Inactive 3-hydroxy-3-methylglutaryl-coenzyme A reductase in broken cell preparations of various mammalian tissues and cell cultures. Preincubation of broken cell preparations from a variety of tissues and cell cultures resulted in an apparent increase in the level of 3-hydroxy-3-methylglutaryl-CoA reductase activity. However, apparent activation of the reductase in mouse liver, hepatomas and primary liver cell cultures was attributed largely to the loss, during the preincubation period, of an interfering enzyme, 3-hydroxy-3-methylglutaryl-CoA lyase. Among non hepatic cells and tissues (which did not contain appreciable lyase activity) the proportion of latent reductase was high in sonicates of fetal brain and in L cells and was independent of the level of total enzyme activity present. Activation of the reductase was blocked by hydroxymethylglutaryl-CoA and NADPH as well as by KF so that activation did not occur under the conditions of the enzyme assay. The enzyme was activated slowly at 4 degrees C, so that partial activation of the latent form occurred during isolation of the microsomal fraction by differential centrifugation. The reductase present in sonicates of cells with either a high or low proportion of the latent enzyme was inactivated by incubation with ATP and Mg2+. Suppression of reductase activity in L cell cultures by treatment with 25-hydroxycholesterol and an age-related decline in brain enzyme activity did not involve reversible conversion of the reductase to an inactive form.


The interference of plasmic degradation products of human crosslinked fibrin with clot formation. The role of plasmic degradation products of human crosslinked fibrin on polymerization of fibrin monomer and clot formation was studied. Both reactions were inhibited by Fragment DD, which formed a complex with fibrin monomer in a molar ratio 1 : 1. The rate of polymerization was slightly increased by Fragment E but it was not affected by (DD)E complex and Fragment A. Approximately the same amount of fibrin was formed in the presence and absence of Fragments A, E and the complex. It was concluded that of the degradation products of crosslinked fibrin, only Fragment DD is a potent anticoagulant at physiologic pH. The (DD)E complex is inert and Fragments A and E have only marginal effects.


Conformational features of bradykinin. A circular dichroism study of some peptide fragments and structural analogues of bradykinin. The vasoactive hormone bradykinin, its N-and C-terminal fragments and some structural analogues were studied by Circular Dichroism. Conformational features of the peptide can be detected by comparative analysis of the various CD spectra recorded as a function of aqueous pH, solvent and temperature. It is shown that the two biologically essential arginine residues (Arg1 and Arg9) are important for the specific folded bradykinin conformation. Differences between bradykinin, its fragments and analogues become clearly established in conformational terms, and are discussed in relation to the biological activity of these peptides.


[Role of proton motive force in the infection of E. coli K-12 cells by bacteriophage T4]. It was shown that infection of E. coli cells by phage T4 is suppressed, when the cells are treated by oxidative phosphorylation uncouplers. The inhibiting effects of the uncouplers manifest themselves at the stage of phage DNA entry into the cells. Study of the E. coli cells devoid of their H+-ATPase activity due to mutation showed that the infection is suppressed by a switch-off of the respiratory chain, the only generator of the proton motive force (PMF) in mutated cells. Infection of the E. coli cells containing intact H+-ATPase occured even in the case when the respiratory chain activity was inhibited. The kinetic studies showed that generation of PMF is necessary during phage DNA transport into the cells and is indispensable for phage DNA entry into bacterial cells.


[Peroxidase activity of hemoglobin, modified at the carboxylic groups of heme and amino acids]. The effects of modification of heme carboxylic groups by omega-aminoenantic acid and L-phenylalamine on the peroxidase activity of hemoglobin were studied. For this purpose the peroxidase activities of the original compounds--hemin, hemin-aminoenantic acid, hemin-phenylalanine and hemoglobins prepared from the hemin and globin compounds--hemoglobin, aminoenantyl-hemoglobin and phenylalanine hemoglobin--were determined. The dependence of the peroxidase activity of these compounds on their concentrations and pH was analyzed. It was shown that 40--50% modification of the heme carboxylic groups by amino acids decreases the peroxidase activity of the modified hemins and that of modified hemoglobins reconstructed from these hemins and globin. A decrease of the catalytic activity of the hemoglobin derivatives is due to a lower peroxidase activity (as compared to hemin) of the modified hemins. It is thus concluded that the amino acid modification of the carboxylic groups of heme does not affect the heme-protein interactions in the hemoglobin molecule.


[Effect of sodium dodecyl sulfate on biological membranes]. Effect of sodium dodecyl sulfate (SDS) on yeast cells resulting in the cellular proteins release from the cells was examined. The influence of pH, ionic strength, the agent concentration and the cells lipids content on the proteins extraction degree was studied. It is assumed that the proteins release is due to the plasma membrane solubilization under the surfactant treatment. It is established that the mechanism of the monimer--membrane interaction and that of the micelle--membrane interaction are likely to be different. It is shown that micelle-like surfactant--protein complexes can solubilize the membrane as well as the SDS micellies. It is found an increase of the ionic strength decreases the efficiency of the SDS action probably due to an increase of the micelle volume. It is assumed that the efficiency of the SDS action on the cell is limited particularly by the ratio of cell wall pere size to the size of micelles formed by the agent in the solution. It is shown also that removal of the largest portion of the lipids from the yeast cell favours an increase in the efficiency of the SDS extractive action at the agent concentrations larger than the critical micelle concentration value probably due to the corresponding decrease of the amount of the cellular components which must be solubilized by the surfactant micelles.


[Glucose induced transport of H+, K+ and lactate in the cells of Acholeplasma laidlawii]. Currents of H+ and K+ ions initiated under the effect of facultative anaerobe of A. laidlawii, valinomycin, nigericin and carbonyl cyanide chloride phenylhydrasone on the membrane are recorded by means of the system of cation sensitive electrodes. It is stated that during glucose transport a redistribution of H+ and K+ and lactate between the cellular content and the medium takes place. Preincubation of cells with 3-o-methyl-D-glucose essentially decreases the effect of induced ion transport. It is concluded that ion transport observed in the experiments with glucose is related not to glucose transport through the cell membrane but to the release of its metabolic products from the cells.


[Dependence of pepsin conformational states on pH and temperature]. A conformational behaviour of pepsin depending on pH and temperature was studied by circular dichroism, differential UV-spectroscopy, calorimetry and enzymatic hydrolysis kinetics. A subtile conformational transition of the enzyme accompanied by changes in the physico-chemical and enzymatic properties of the protein was observed within the temperature interval of 15--40 degrees and within the pH range of 1,1--5,6. The range of pepsin heat denaturation was studied. A diagram of pepsin conformational states under different values of pH and temperature was built.


[Soluble high molecular weight derivatives of pancreatic inhibitors. Kinetic-thermodynamic studies of inhibitors linked to differently charged matrices]. The effects of electrostatic charge of the matrix on the pH-dependence of interactions of commercial trypsin with preparations of pancreatic inhibitor modified by soluble polysaccharide coupling were studied. It was shown that the rate constants of trypsin association with native and modified pancreatic inhibitor preparations as well as the rate constants of dissociation of their complexes and, consequently, the inhibition constants are identical. The invariability of the rate constants for the association reaction after the increase in the molecular weight of pancreatic inhibitor may be probably accounted for by the fact that the limiting step of a stable trypsin-inhibitor complex formation is not controlled by diffusion. Thermal denaturation of pancreatic inhibitor preparations modified by binding to polysaccharides (pH 4.7--8.0, 97 degrees C) suggests an essential role of the negative charge of matrix in stabilization of the protein inhibitor globule.


[Intermediate products of the RNA-ligase reaction]. The first and the third steps of the RNA-ligase reaction were studied. It was shown that the first step of the reaction consists in a formation of an enzyme-adenylate complex. The optimal conditions for this formation were established. Effects of acids, alkali, hydroxylamine and snake venom phosphodiesterase on the complex suggest that the linkage between the protein and adenylic acid may be of a phosphoester or phosphoamide type. Using synthetic adenylic acid pyrophosphates and mononucleotides (oligonucleotides) the RNA-ligase reaction was shown to involve intermediate pyrophosphates. It was found that the simplest pyrophosphates capable to bind to oligonucleotides in the absence of ATP are adenylic acid pyrophosphates, both of ribo- and deoxyribomononucleotides. The RNA-ligase reaction may be used for elongation of oligonucleotides by one definite mononucleotide or for incorporation of the label into the 3'-end of the polynucleotide chain.


[Isolation, purification and study of the stability of the soluble hydrogenase of Alcaligenes eutrophus Z-1]. Soluble hydrogenase was isolated from the hydrogen-oxidizing bacterium Alcaligenes eutrophus Z-1 and purified to electrophoretical homogeneity. The purification procedure included fractionation by ammonium sulfate, ion-exchange chromatography on DEAE-cellulose and gelfiltration through Ultragel AcA-34. The resulting preparation had a specific activity of 25 mkmoles H2.min-1.mg of protein as measured by the rate of hydrogen evolution from sodium dithionite-reduced methyl viologen. The enzyme has a molecular weight of 200,000 and is made up of two subunits with mol. weights of 30,000 and two subunits with mol. weights of 65,000. The effects of pH, oxidants and reducers, as well as aerobic and anaerobic conditions on the hydrogenase preparations inactivation kinetics in intact cells and in a highly purified state were studied. The kinetic data suggest a possible existence of two enzyme forms differing in their activities and stabilities to denaturating influences.


[Comparative study of hydrolysis of methyl esters of N-arylsulfonyl-valyl-arginine by thrombin and trypsin]. The esterase action of thrombin and trypsin on N-arylsulfonyl-valyl-arginine methyl esters was studied. The values of Km and kcat under steady-state conditions at pH 8,5 were determined. It was shown that the nature of the arylsulfonyl group does not affect the kinetic parameters of the reactions under study. The Michaelis constants of the thrombin-catalyzed reactions appeared to be one order of magnitude lower than the Km values of the corresponding TAME analogs.