Interactions of gastric blood flow, barrier breaker, and hydrogen ion back diffusion during ulcer formation in the rat. A rat gastric chamber preparation was used to determine the interrelations between H+ back diffusion, mucosal blood flow, the area of mucosa treated with bile salts and ulcer formation. The rate of H+ back diffusion was found to be dependent on both mucosal blood flow and the area of mucosa exposed to bile salts. It is concluded that, although H+ back diffusion is a convenient measure of barrier permeability under conditions of normal mucosal blood flow, during mucosal ischemia this parameter fails to reflect permeability because of decreased mucosal clearance of H+. These findings would explain apparent controversy whereby ulceration occurs under shock conditions in the absence of apparent H+ back diffusion.


Lateral ventral (spigelian) hernias in infants and children. Lateral ventral hernias are found in the spigelian fascia lateral to the junction of the arcuate and semilunar lines and below the level of the umbilicus. Three cases are reported in infants and children. Diagnosis depends on finding an unusual mass on the anterior abdominal wall and palpation of the rim of the hernia defect upon reduction of the mass. A spigelian hernia may be misdiagnosed easily if the examiner is unaware that it can occur in childhood. The edge of the defect must be outlined while the child is straining, prior to induction of anesthesia, as the rim may be difficult to find when the patient is asleep.


The efficiency of diamphenethide in controlling natural infestations of Fasciola hepatica in lambs. The activity of diamphenethide against natural infestations of Fasciola hepatica in fattening lambs was assessed on a farm with a noted history of fascioliasis. The main experimental groups, two treated and one control, grazed known contaminated pastures for a twelve week period after which they were removed to fluke-free areas to await slaughter. Three groups of four tracer lambs were each grazed for three successive four week periods with the main flock to determine continuity of infection. A single treatment with diamphenethide at 110 mg/kg bodyweight given when the sheep were removed from the trial area was 96.7% effective against the sub-chronic disease which had developed by this time. Three treatments at 110 mg diamphenethide/kg bodeweight given at intervals of four weeks during the grazing period proved to be 100% effective in preventing the establishment of the disease. Significant benefits in terms of weight gain were exhibited by the treated lambs.


Arbovirus isolations from mosquitoes: Kano Plain, Kenya. Arbovirus isolation attempts on 324,486 mosquitoes captured over a four-year period on the Kano Plain, Kenya, yielded 15 isolates including Pongola (six strains), Ilesha (three strains), Germiston (two strains), Sindbis (one strain), Barur (one strain) and two viruses which could not be characterized. Mansonia uniformis, Anopheles gambiae and Culex antennatus constituted 70% of the total collection and accounted for all of the isolates except one, which came from Anopheles funestus.


Congenital testicular lymphangiectasis.

Congenital testicular lymphangiectasis. Testicular lymphangiectasis are described for the first time in a patient with bilateral inguinal cryptorchidism. A great number of irregular lymphatic channels was observed within the parenchyma and the tunica vasculosa in both testes. Large and numerous anastomosis between the lymphatic vessels of these two areas could also be seen. The MTD and the TFI of the left testis were normal. Both parameters were very low in the right testis. The association of this fact with the greater development of the lymphatic vessels in this testis strongly supports the idea that testicular lymphangiectasis interfere mechanically with the testis tubular development.


Preservation of human granulocytes. III. Liquid preservation studied by electronic sizing. Human granulocytes were isolated from blood either by counterflow centrifugation in a Beckman JE-6 rotor or by sedimentation of the red blood cells with dextran and centrifugation of the granulocyte-rich plasma. The stability of the granulocytes was assessed during storage in the liquid state by measurements of granulocyte loss, volume distribution characteristics, and ability to produce fluorescein in their cytoplasm and to exclude ethidium bromide from their nuclei. After storage at 4 C for 2 days in phosphate-buffered saline with a pH of 7.1 containing 5 g/dl human albumin + 0.46 g/dl dextrose or 1 g/dl Physiogel + 0.46 g/dl dextrose, the granulocytes were adequately preserved from the in vitro measurements.


Carcinoid tumour of the thymus. A case report including discussion of the morphological diagnosis and the cell of origin. This report concerns a 49 year old asymptomatic male who had a mediastinal mass demonstrated on routine radiography. A large encapsulated tumour composed of small regular cells arranged in clumps and acini with fine vascular stroma was removed. The differential diagnosis on routine H&E section included parathyroid tumor, medullary carcinoma arising in ectopic thyroid tissue, epithelial thymoma or carcinoid tumor of the thymus. The presence of compressed thymic tissue around the tumor, and of argentaffin granules together with the electron microscopic appearance characteristic of the "enterochromaffin" or "APUD" group of cells allowed the diagnosis of carcinoid tumor of the thymus to be made. Electron microscopy showed that the cell cytoplasm contained electron dense membrane bound granules, together with bundles of microfilaments. Vesicles of smooth surfaced reticulum were present but rough surfaced reticulum was inconspicuous. No desmosomes were demonstrated. Special stains for amyloid and glycogen were negative.


The effect of agitation of stored human blood on microaggregate formation. Microaggregates (MA) composed of platelets and white blood cells form during the storage of human blood. These particles are believed to be a cause of pulmonary insufficiency in patients receiving massive blood transfusions. The present controlled study determined the effect of constant gentle agitation of CPD-anticoagulated blood, during storage at 4 C, on the formation of MA. Using a Model T Coulter Counter, it was found that agitated blood contained significantly lower volumes of MA at 7 and 14 days than did nonagitated controls. However, significant elevations, above control levels, of plasma free hemoglobin, lactic dehydrogenase, and potassium indicated significant injury to cellular components of agitated blood. This study raises serious doubts concerning the potential clinical usefulness of blood agitation during storage to prevent MA formation.


[Some properties of Mg 2+, Ca 2+-AtPase of brain microsomal fractions and synaptic plasma membranes in the solubilized state]. Digitonin-solubilized Mg2+, Ca2+-ATPases of the brain microsomal fraction and synaptic plasma membranes are activated against a background of 6.0 mM Mg2+ by 0.001 and 0.03 mM of the added Ca2+ and manifest maximum activity at temperatures or 40-45 and 35-42 degrees C, respectively. The enzymes of the initial membrane preparations are activated by the same concentrations of the added Ca2+ and are characterized by the same temperature dependence. The values of pH-optima of the enzyme activity of both solubilized preparations are 7.0-7.3 and in comparison with pH-dependence of the initial membrane preparations are shifted towards acid side.


The disposition of phentermine and its N-oxidized metabolic products in the rabbit. 1. When phentermine was injected intraperitoneally to rabbits, 77% of the dose was excreted in the urine within 2 days; N-oxidized metabolic products accounted for 62% dose. Major excretion products were N-hydroxyphentermine (28% dose), conjugated N-hydroxyphentermine (32% dose) and unchanged phentermine (16% dose). 2. Similarly injected N-hydroxyphentermine was excreted (62% dose) in the urine in 2 days; only 4% dose was recovered unchanged. Major routes of metabolism of N-hydroxyphentermine was conjugation (36% dose) and reduction to the parent amine (15% dose). Conditions for hydrolysis of urine to liberate N-hydroxyphentermine from its conjugates were studied; N-hydroxyphentermide decomposes in strong acid. 3. Only 10% of injected alpha, alpha-dimethyl-alpha-nitroso-beta-phenylethane was excreted in the urine in 2 days. 4. N-Oxidations is the major pathway of metabolism of phentermine in rabbits; the present results suggest that some biological activity may be mediated by the pharmacologically active N-hydroxyphentermine.


[Neonatal depression in the absence of azidose (author's transl)]. In addition to those children delivered by cesarean, a noticeable number of premature infants show a neonatal depression without intrauterine or neonatal acidosis. The reasons for this are only partially known. Anesthesia, on the one hand, and the underdeveloped brain, on the other hand, are held responsible. The prognosis of these children is relatively serious. On the one hand, they show a frequent incidence of neurologic disorders, or an increased rate of deformity on the other which although without immediate intrauterine consequences, will render extrauterine life substantially more difficult, if not impossible.


Studies on glutamic-oxalacetic (GOT) and glutamic-pyruvic (GPT) transaminases of swine kidney worm Stephanurus dentatus (Diesing, 1839). I. Assay and general properties. Biochemical studies on the two transaminases GOT and GPT of swine kidney worm Stephanurus dentatus have been made. GOT has been found much more active than GPT. Enzyme activities are based on the formation of oxaloacetate (GOT) or pyruvate (GPT) from aspartic acid and alanine respectively with oxoglutarate. A linear relationship is observed between the enzyme concentration and activity. GOT shows a maximum activity at pH 8.0 and Michaelis constant 9 X 10(-3) M for male and 2.9 X 10(-3) M for female. GPT has an optimum pH of 7.5 and a Michaelis constant 19 X 10(-3) M for male and 8 X 10(-3) M for female. The optimum temperature for both GOT and GPT was 60 degrees C.


[The significance of serum gamma-glutamyl transpeptidase determination in the differentiation of icterus of various etiologies]. In a group of 29 patients with mechanical icterus and a control group of 38 patients (acute, chronic hepatitis and hyperbilirubinemia) there have been followed up alkaline phosphatase, leucinaminopeptidase and gammaglutamil transpeptidase activities aiming to evaluate the value of gamma GT in differentiation of icterus of various etiology. Comparing the values of the examined cholestase enzymes in patients of obstructive icterus it could be seen that gamma GT was positive in the largest percentage of the diseased, (69,6%), then APH (89,7%) AND LAP (86,3%). Gamma GT has shown to be selective test for obstruction of hepatobiliary tract but it is not selective for discovering cause of obstruction. It can be explained by the fact that activity of gamma GT could be found both in mechanical icterus caused by calculosis or tumours (with a very high activity) and in patients with hepatitis with cholestatic component, although in the decresed activity.


The stability properties of golden beet and red beet pigments: influence of pH, temperature, and some stabilizers. Golden beet (Beta vulgaris var. lutea) color was found to contain 8 components, of which the most important seemed to be vulgaxanthin-I and -II. Purified vulgazanthin -I solutions are easily degraded at high temperatures. The stability is, however, strongly dependent on pH: the stability is best at pH values between 5 and 6, the poorest at low pH values, but at pH 7 the stability is better than that of betanin. Vulgaxanthin-I is more stable in the raw extract than in a purified solution. EDTA, even in ppm amounts, was found to improve the stability of betanin. The effect was dependent on pH value, being most efficient at pH 2.0 and 5.0. Stability of vulgaxanthin-I was not improved by EDTA. Tannin did not show stabilizing influence. Ascorbic acid was found to have an unfavorable influence on stability, especially in the case of betanin.


[Dialysis technic for regulation of the internal milieu in acute poisoning]. In 72 patients with acute poisoning (15 patients without unconsciousness with 80% of acute renal and hepatic failure, and 57 patients with unconciousness with 10.5% of acute renal and hepatic failure) the significance of the continuous prolonged haemodialysis lasting on an average 12 hours, respectively 18 hours, with the maximum of 33 and 82 hour, respectively. The duration of the haemodialysis was in close correlation with the pH-value of the blood and the duration of the unconciousness. One half of the unconscious patients suffered from dehydration (of them up to 40.3%), which was valuated by the height of the central venous blood pressure. It was also referred to the necessity of the correction of the potassiaemia and glycaemia.


[Current status of treatment of parasellar meningiomas]. For 302 patients presenting the respective indication in the years 1968 to 1972 the operation lethality amounted to 24.8%. Then the microtechnique including the operation microscope were introduced. Out of 47 cases of tuberculum sellae meningiomas one patient died; out of 42 patients suffering from median meningiomas of the wing of the sphenoid bone nine died. Lethality decreased by one half, radical surgery increased from 50 to 80 to 85%. A contraindication for surgery was not considered to be present even with a bad general condition. Post-operative control: cerebrospinal pressure, intracerebral pressure, pressure of the interstitial fluid, local and regional blood supply, pO2 and pCO2, electrolyte balance, pH, osmolality. This allowed also to record conditions of underpressure.


[Genital disorders that influence fertility and potency. Screening methods]. It is recommended to introduce screening for fertility and potency determination to disclose genital disturbances the correction of which must be done in childhood or early age. Cryptorchism, undescended testicles or their hypoplasia, hypogonadism, hypogenitalism, hypospadias and finally varicocele are easily detected by screening at school, on military enlistment or by the sports physician.


Changes in serum TSH level after intraventricular injection of various neuromediators in rats. The influence of various neuromediators on pituitary TSH secretion in rats has been investigated. Noradrenaline 50 microgram/rat, dopamine 50 microgram/rat, serotonine-creatinine-sulphate 100 micogram/rat, gamma-aminobutyric acid 100 microgram/rat, pilocarpine 1 mg/rat, histamine 100 microgram/rat were administered into the lateral ventricle of the brain. All agents were dissolved in Parker's fluid. Two control groups of animals were given Parker's fluid and subjected to surgical manipulations, respectively. Plasma TSH level was estimated after 30 min by means of radioimmunoassay. The increase in the TSH level was observed after the injection of serotonine and noradrenaline (4.0 and 3.1 ng/ml, respectively) as compared with control group (0.7 ng/ml).


ATP-induced release of vasopressin associated with phosphorylation of isolated bovine neurohypophyseal secretory granule membranes. Secretory granules isolated from ox neurohypophyses released their content of vasopressin in the presence of ATP and Mg2+. A half maximal ATP concentration of 0.25 mM was found. Ca2+ was not necessary for the effect. High concentrations of ADP, AMP and ITP were shown to mimic the effect of ATP. Utilizing this effect of ATP combined with iodonitrotetrazolium treatment to make mitochondria heavier, a method is described to obtain granule "ghosts" in a purified form. They were shown to be phosphorylated when granules were incubated with [gamma-32P] ATP.


Evaluation of the usefulness of serum gamma-glutamyl transpeptidase levels in the management of haematological neoplasia. The levels of serum gamma-glutamyl transpeptidase (GGT) and, when appropriate, alkaline phosphatase (AP) and 5'-nucleotidase (NTD) have been measured as a routine in 276 patients with malignant haematological diseases during a 26-month trial period. GGT levels add no prognostic information to the routine haematological surveillance of leukaemia. Polychemotherapy does not appear to be an inducer of liver drug-metabolising microsomal enzymes. Polycythaemia rubra vera, myelofibrosis and chronic lymphocytic leukaemia may cause little change in GGT, AP and NTD levels despite marked hepatomegaly. A raised GGT in Hodgkin's disease and non-Hodgkin lymphoma is generally associated with active and widespread disease, but not necessarily a sign of malignant tissue in the liver. The elevations of GGT in myeloma may be secondary to liver infiltration though this group merits further detailed study.


"False positive" acidified serum test in a preleukemic dyserythropoiesis. In a case of preleukemic dyserythropoiesis, in vitro red cell lysis tests showed a positive acidified serum test whose characteristics are described. The positive acidified serum test occurred in 10 normal sera, in 1 serum with complete deficiency of the fourth component of complement and in 1 serum with complete deficiency of the second component of complement. The test was found negative with 2 hyperlipemic sera. The other in vitro red-cell lysis tests were negative. The results of the in vitro lysis tests were different from the results obtained in paroxysmal nocturnal hemoglobinuria and congenital dyserythropoietic anemias.


Pharmacokinetics of 2-pivaloylindan-1,3-dione in dogs. The pharmacokinetics of 2-pivaloylindan-1,3-dione (Pival) after administration of a single dose was investigated in dogs. The plasma concentration graph can be described by a one compartment open model. The drug is fairly well adsorbed (67%) and the excretion is extremely slow with a half-life of nearly five days. These kinetics can be explained by the physiochemical properties of the drug, such as strong binding to canine plasma protein, high lipophility and the lack of transformation to polar metabolites. A single oral dose of 5 mg/kg must be regarded as lethal to dogs and the LD50 is assumed to be approximately 4 mg/kg in this species.


Minoxidil in severe hypertension.

Minoxidil in severe hypertension. Minoxidil has been administered to 16 patients with severe hypertension and renal failure. In every patient the indication for minoxidil treatment was resistance to conventional drugs. The final dose of minoxidil was 2.5--30 mg (average 20) and it was combined with a beta-blocking agent and a diuretic (or dialysis). The therapy was given for 1--27 months (average 12). The average supine BP fell from 200/130 to 164/96 mmHg and the upright BP from 200/120 to 152/90 mmHg. No hypotensive reactions occurred. In most patients the progression of hypertensive organ changes was arrested. No major vascular complications have occurred during the 16 years of treatment. Prickling of the skin and hirsutism were common side-effects. The other side-effects observed were oedema in five patients and development of latent diabetes in three. In four patients minoxidil treatment was discontinued for following reasons: successful reconstruction of the renal artery after stenosis, renal transplantation, severe oedema and hirsutism. The risk of hirsutism is a contraindication to prolonged minoxidil administration in most femal patients. Minoxidil is especially indicated in uncontrolled renal hypertension.


Histamine H2-receptor antagonists.

Histamine H2-receptor antagonists. Development of histamine H2-receptor antagonists has enhanced the understanding of histamine physiology and pharmacology. The effect of H2-receptor antagonists on gastrointestinal physiology has been studied extensively. These compounds inhibit gastric acid secretion in response to all known secretagogues and, in contrast to anticholinergic drugs, markedly inhibit food-stimulated acid secretion in duodenal ulcer patients. The relative roles of H2-receptor antagonists, anticholinergic drugs and antacids in the treatment of duodenal ulcer remain to be defined. Cimetidine currently is under investigation for the treatment of duodenal ulcer, gastric ulcer, reflux esophagitis, gastrointestinal bleeding and hypersecretory states. Although the long-term safety of cimetidine has not been established, in short-term clinical trials there have been no significant subjective or objective side-effects. Assuming that toxic effects do not develop, H2-receptor antagonists should improve the treatment of acid-peptic disease.


Biodistribution of salicylates: a clue to the understanding of some effects and side effects. According to our concept that biodistribution of salicylates (SA) might be an important factor of their mode of action the distribution of sodiumsalicylate (NaSA) and acetylsalicylic acid (ASA) was investigated in vivo and in vitro. It was shown that particular high concentrations of both compounds were found in vivo in the glandular stomach region and the kidney tubules of rats as well as in inflamed tissue. Quantitative studies showed that in chicken, in inflamed joints, higher concentrations as in plasma and about three times higher concentrations as in control joints were observed. In vitro studies using human erythrocytes suspended in buffer of different pH showed that added NaSA accumulated in these cells when the environmental pH was lowered from 7.4 to 6.8. From these results it is speculated that SA being weak acids accumulate within cells which are surrounded or bordering acidic fluids. This is the case in the stomach, the kidney and the inflamed tissue. This is where SA exert their effects and side effects.


Renal excretion of fluid, electrolytes and hydrogen ions before and during diamox administration in healthy Nigerians. Twenty-four urine specimens of eleven healthy Nigerians taken during the cool rainy season, were examined before and during diamox (acetazolamide) administration. Mean control volume-output was 1.4 litres containing 109.5 mEq Na+, 46.6 mEq K+ and 120 mEq Cl- ions. Total renal acid excretion was 36.82 partitioned as 14.72 mEq titratable buffer-acid and 22.1 mEq ammonia. During diamox intake, volume-output increased to 2.56 litres with Na+ and K+ contents of 222.4 and 107 mEq respectively, whilst Cl- ions, fell to 2.6 mEq. Daily renal acid excretion was reduced by 25% and titratable buffer acid was absent in most samples but ammonia excretion showed a wide scatter with a range of 6.5-60.8 mEq. The probable explanations for the lower potassium and total acid excretions during the control period as well as the pattern of ammonia excretion during diamox therapy are discussed.


Orungo virus, an orbivirus from Africa: effects of physical and chemical agents. Orungo virus is relatively sensitive to lipid solvents and sodium deoxycholate. It is readily inactivated at 37 degrees C and above, by u.v. irradiation, BP and formalin. Orungo virus is ph 3.0 labile, but stable at ph range of 5.0-7.0. Multiplication of Orungo virus in BHK-21 cell cultures is not affected in the presence of 5-iododeoxyuridine.


Pulmonary function following feeding in low-birth-weight infants. Determination of functional residual capacity, arterial gas tensions and pH, and arterial-alveolar differences was carried out in a group of nondistressed premature infants to determine if significant alterations in lung volumes or ventilation perfusion relationships sufficiently large to cause cyanosis could be detected after feeding. The only statistically significant changes observed in these parameters were immediately following feeding, a fall of .01 pH units and a fall of 6 mm Hg in arterial oxygen tension; and 15 minutes after feeding, a fall of .01 pH units and a rise in arterial PCO2 of 3 mm Hg. In addition, significant falls in peripheral blood flow were observed five minutes after feeding in association with marked elevations in peripheral vascular resistance. The mild impairment in pulmonary function in association with the more profound changes in limb blood flow are consistent with other studies in premature and full-term infants. These data suggest that cyanotic attacks following feeding may have a complex etiology and require an aggressive multisystem approach for adequate diagnosis and therapy.


Toxic epidermal necrolysis from graft-vs-host disease. Occurrence in a patient with thymic hypoplasia. An infant with diarrhea, failure to thrive, and a seborrhea-like skin eruption was thought to have fatal familial Leiner's syndrome. Treatment with nonirradiated plasma was followed by graft-vs-host disease and fatal toxic epidermal necrolysis; thymic hypoplasia was found at autopsy. Accurate diagnosis of immunodeficiency syndromes is essential to avoid potentially harmful therapy.


Diagnosis and treatment of diffuse ileojejunitis. Thirteen patients with diffuse ileojejunitis have been diagnosed and treated by us over the past ten years. The disease bears close resemblance to Crohn's disease and may represent a variant of it. No clearcut relationship to celiac sprue was observed in this group of patients. Therapeutic success was obtained in the majority of patients treated with the anti-inflammatory drugs, sulfasalazine and steroids, with four patients requiring resectional surgery, all others manageable by nonsurgical means. There was no mortality in this series of patients.


H2 receptor antagonists.

H2 receptor antagonists. The uses of the histamine H2 receptor antagonists (H2 antagonists) in the management of peptic ulcer disease and the Zollinger-Ellison syndrome are reviewed. Drugs included in the discussion are burimamide, metiamide and cimetidine. The secretion and the pharmacology of the H2 antagonists are described. A discussion of the clinical use of the H2 receptor antagonists in the treatment of gastric hyperacidity and hypersecretory states, and the side effects encountered is presented. Cimetidine is effective in the treatment of duodenal ulcer, and its use appears promising in Zollinger-Ellison syndrome and gastric ulceration.


Pharmacists' knowledge and attitudes regarding monitoring the behaviors of children with learning disabilities. The attitudes and knowledge of pharmacists in Nebraska regarding children's learning disabilities were surveyed by questionnaire. One hundred thirty-three pharmacists responded. The results were analyzed according to type of practice (community, chain and hospital) and age (20--30 years, 31--40 years, 41--50 years, and 51 years and over). The results indicate that pharmacists are being asked about learning disabilities. Overall, 53% of the pharmacists responding had been asked about learning disabilities by parents, physicians, teachers or others. No significant interactions were found between the type of practice and age, and no significant main effect for type of practice was found in the ANOVA results for knowledge scores or attitude scores. Even though there were no differences owing to age in pharmacists' knowledge of learning disabilities, age was found to have a significant effect on attitudes toward increased pharmacist involvement in monitoring behaviors of learning-disabled children; the younger groups had a more positive attitude. The pharmacist seems to have the knowledge and attitudes required to become more actively involved in monitoring the medications and behaviors of children with learning disabilities.


Development of neurohumoral control of fetal, neonatal, and adult cardiovascular functions. Neurohumoral control of fetal, neonatal, and adult cardiovascular functions have been reviewed. Resting fetal heart rate remains fairly constant but neonatal heart rate declines progressively, reaching adult levels within six to eight weeks; systemic arterial pressure rises while pulmonary pressure falls to adult levels within the first week after birth. Sympathetic and parasympathetic control of circulatory functions matures at different rates during fetal and neonatal development; the sympathetic system becomes active earlier in fetal life than does the parasympathetic system. After birth, the parasympathetic tone of the resting heart rate rises to adult levels while adrenergic tone decreases. Despite changing autonomic activities, resting heart rate is set at given levels through alterations in intrinsic control. In the fetus, peripheral circulation is under neurohumoral tone of increasing magnitude; after birth, neurohumoral tone declines progressively, reaching levels comparable to those of adult nonpregnant sheep. Fetal cardiovascular response to neurotransmitters increases with age because of maturation of the effector system. The pulmonary bed responds primarily to acetylcholine whereas the systemic circulation responds to norepinephrine. After birth, the neonatal cardiovascular system becomes four to five times more sensitive to the action of neurotransmitters mainly because of closure of vascular shunts and elimination of umbilicoplacental circulation. In the neonate and adult, the pulmonary vascular bed loses its reactivity to neurotransmitters.


Immune complexes of hepatitis B surface antigen in the pathogenesis of periarteritis nodosa. A study of seven necropsy cases. In 7 unselected necropsy cases of clinically diagnosed periarteritis nodosa, the detection of hepatitis B surface antigen (HBsAg) and hepatitis B core antigen (HBcAg) in the cytoplasm and nuclei of hepatocytes indicated an ongoing infection with hepititis B virus (HBV). In all these cases histologic changes found in the liver varied from "minimal" to chronic aggressive hepatitis. In all the cases, deposits of HBsAg, immunoglobulins, beta1C-globulin and C1q were detected in vascular lesions. That these deposits could represent HBsAg-anti-HBs immune complexes was supported by demonstrating their strong binding of guinea pig complement and by the successful elution of all HBsAg and part of the immunoglobulin from these deposits by treatment with buffers known to dissociate antigen-antibody bonds but not with phosphate-buffered saline, pH 7.6 (PBS). Glomerulonephritis associated with these immune complexes was found in 6 cases. The presence of larger masses of HBsAg immune complexes, chiefly in recent insudative and fibrinoid vascular lesions, their lesser amounts in lesions undergoing involution, and their absence from healed lesions strongly suggest that these complexes play a primary role in the pathogenesis of acute vascular damage in periarteritis nodosa.


Tumor induction in host-versus-graft disease. I. Clinical and pathologic features. Perinatal C57BL/1 mice given injections of (SJL/J X C57BL/1)F1 spleen cells developed a highly lethal runting syndrome, designated host-versus-graft disease (HVGD). The mortality was related to the dosage of F1 cells. Acute pathologic changes resembled those occurring in parent leads to F1 graft-versus-host disease (GVHD), except for more pronounced plasmacytosis. Mice suffering from HVGD recovered clinically with no sequelae except for a slight increase in  the incidence of lymphomas over control mice. Such mice were hyperreactive to F1 cells utilized to initiate the original HVGD syndrome. Most of the tumors developed in those animals receiving the initial injection of F1 spleen cells within 24 hours of birth. Tumor incidence was unrelated to the clinical severity of HVGD. By contrast, GVHD in the same strain combination resulted in a much higher incidence of lymphomas in a much shorter time. Parental strain cells were detectable in the F1 hosts up to the time of tumor development. HVGD has a low tumor induction potential; GVHD has a high tumor induction potential.


Association of increased pHi with calcium ion release in skeletal muscle. The pH difference across the cell membrane of frog sartorius muscle cells was measured with the distribution of 5,5-dimethyl-2,4-oxazolidine-dione (DMO) as the marker. Depolarization of the muscles to values at or below the contraction threshold caused by elevating external potassium up to approximately 20 mM resulted in an internal alkalinization. The change was smaller with superthreshold depolarization (20--30 mM [K+]). The alkalinization was blocked by agents that block calcium release from the sarcoplasmic reticulum (procaine and dantrolene sodium). Other agents that cause calcium release (caffeine, theophylline, and quinine) were found to give alkalinization when tested at concentrations just below the contracture threshold. Increased acidification of the extracellular medium was associated with the internal alkalinization. The data were interpreted as indicating the presence of a calcium-stimulated H+ and/or OH- ion transport system in the muscle membrane.


Na+ transport by mammalian stomach.

Na+ transport by mammalian stomach. Gastric mucosas from newborn pigs (0--20 days) and rabbits (0--20 days) were used for in vitro investigation of active Na+ transport during resting (no HCl secretion) conditions. As measured with 22Na+, these tissues actively absorb Na+ from the mucosal-to serosal (m-t-s) bathing solution during both open-circuit and short-circuit current (Is) conditions. In the nonsecreting state, net Na+ transport accounts for 40--60% of Isc. The remaining current is provided by net s-to-m flux of Cl-. Amiloride (2-5 X 10(-5) M) in the mucosal solution abolishes this active Na+ transport by inhibiting m-to-s fluxes of Na+ (JNams). In vivo-in vitro experiments showed that active Na+ transport is a normal function of the resting mammalian stomach. Decreasing pH of the mucosal solution below pH 5 reversibly causes decreased current-generating capability of the tissue. Pretreatment of the tissue with amiloride abolishes this pH effect. The implication is that the low pH affects the Na+-entry step into cells. "Titration curves" of current vs. pH had an apparent pK approximately 4.0. Ouabain and K+-free solutions both cause decreases in active Na+ and Cl- current. Calculations indicate that a shunt may account for only a small (less than 30%) percentage of total transepithelial conductance.


Norepinephrine: hormone and neurotransmitter in man. To determine whether norepinephrine could subserve a hormonal as well as a neurotransmitter function, norepinephrine was infused for 60 min into each of five normal young men in doses of 0.1, 0.5, 1.0, 2.5, and 5.0 microgram/min. After infusion, the plasma norepinephrine concentration fell with a mean (+/-SD) half-time of 2.4 +/- 0.7 min. The mean (+/-SD) norepinephrine metabolic clearance rate was 3,070 +/- 200 ml/min. The calculated basal plasma norepinephrine production rate was 0.7 microgram/min. The blood pressure and circulating glycerol, acetoacetate, beta-hydroxybutyrate, and glucose (increased) and the heart rate and circulating insulin, lactate, pyruvate, and alanine (decreased) exhibited highly significant parabolic relationships with the steady-state plasma norepinephrine concentrations. However, norepinephrine levels in excess of 1,800 pg/ml were required to produce hemodynamic and/or metabolic effects. Thus, under usual conditions, the biologic actions of norepinephrine can be attributed only to its sympathetic neurotransmitter function. Plasma norepinephrine concentrations do at times exceed 1,800 pg/ml during exercise and during major acute illness. Thus, under conditions of stress, norepinephrine may subserve a hormonal, as well as a neurotransmitter, function.


Acetazolamide and renal ammoniagenesis.

Acetazolamide and renal ammoniagenesis. The effect of acetazolamide on ammonia-producing enzyme systems was determined in vitro at concentrations comparable to those which have been shown to abolish ammonium excretion in vivo. No change in the activity of glutaminase or gamma-glutamyl transpeptidase could be observed at concentrations up to 0.2 mM acetazolamide, and concentrations up to 1 mM were without effect on D-glutamyltransferase activity. Therefore, the effect of acetazolamide to abolish ammonium excretion cannot be explained by an action of the drug to inhibit ammoniagenesis.


Cerebral blood flow in birds: effect of hypoxia. The effect of hypoxia on cerebral blood flow in ducks was investigated by the rate at which arterially injected xenon-133 was cleared from the duck's brain. A two-component clearance curve resulted, which we have attributed to flow through the grey and white matter. Decreasing the arterial oxygen tension (PaO2) to 75 mmHg had no effect on cerebral blood flow. However, decreasing the PaO2 below 75 mmHg significantly increased blood flow to the fast-clearing compartment. The greatest increase in blood flow was seen when the arterial PO2 was below 50 mmHg. At an arterial PO2 of 30 mmHg, the cerebral blood flow to the fast-clearing compartment was increased more than 600% above the normoxic level. The magnitude of this increase is much greater in the duck than has been reported for mammals at roughly equivalent arterial oxygen tensions. The ability of avian cerebral blood flow to increase at moderate levels of hypoxia, plus the magnitude of the increase, may play a role in the exceptional tolerance of birds to hypoxia.


Altered hemodynamic responses to acute hypoxemia in spontaneously hypertensive rats. Conscious spontaneously hypertensive rats (SHR), 5--7 wk old, were studied hemodynamically by the direct Fick procedure to determine whether high total peripheral resistance (TPR) coexisted with increased oxygen consumption (QO2) at an early stage of hypertension development. Since under resting conditions cardiac output in SHR was not significantly different from normotensive controls, the elevated arterial pressure and QO2 were associated with increased TPR. Arterial hypoxemia was induced to reduce oxygen availability and to assess whether increased TPR in SHR could be reversed by this procedure. During hypoxemia, normotensive controls (WKY) responded with increased cardiac output and decreased arterial pressure and TPR. In contrast, arterial pressure and cardiac output fell in SHR; and the increased TPR persisted. QO2 fell in hypoxemic SHR demonstrating that the relationship between total body oxygen consumption and cardiac output was abnormal in young SHR, and that increased TPR in SHR was not dependent on resting levels of QO2 or oxygen availability. Although QO2 was elevated in SHR compared to age-matched WKY, this condition was not essential for maintained elevated vascular resistance.


Controlling the highest lactate dehydrogenase activity known in nature. In the shipjack, Euthynnus pelamis, white muscle appears to possess a powerful anaerobic capacity as well as a significant carbohydrate based aerobic potential. Lactate dehydrogenase occurs at higher activities than found thus far anywhere else in nature and clearly functions in anaerobic glycolysis. Alpha-glycerophosphate dehydrogenase also occurs in unusually high activities and appears to play a role in aerobic glycolysis. Regulation of these two reactions is accomplished by temperature, pH, and creatine phosphate levels. High temperature, low pH, and low creatine phosphate levels all appear to favor lactate dehydrogenase over alpha-glycerophosphate dehydrogenase; low temperature, high pH, and high creatine-phosphate levels, all expected during the quiescent state in this species, and when metabolism in aerobic, all favor alpha-glycerophosphate dehydrogenase activity.


Gastric mucosal defense mechanisms: effects of salicylate and histamine. Some of the recent concepts about the gastric mucosal defense mechanisms against damage by luminal acid and the effects of histamine and salicylate on these mechanisms are reviewed. The mucosal barrier to acid appears to consist of at least two physiologic components: a permeability mechanism and a metabolic mechanism related to cellular bicarbonate production as a result of acid secretion. In the absence of salicylate, histamine appears to exert some protection by affecting both mechanisms, but in the presence of salicylate, histamine's protective effect is limited to altering mucosal permeability. The actions of salicylate on the gastric mucosa are complex, related in part to the concentration of salicylate and the pH of the luminal fluid. The damaging effects of salicylate appear to be related more to the concentration of acid in the lumen than to the lipid solubility of the drug. Salicylate increases permeability regardless of pH; the increase is initially selective for cations and subsequently becomes nonselective, involving both cations and anions. Although both low and high concentrations of salicylate increase mucosal permeability to hydrogen ions, only high concentrations of salicylate affect cellular bicarbonate production.


The neurohumoral response to trauma.

The neurohumoral response to trauma. The bodily response to injury is an integrated and protective mechanism involving all systems, and hence it should be considered as a whole. The nervous system, especially the psychic center of the cerebral cortex, seems to initiate these responses. In the present study the neurohumoral response to trauma was assessed in experimental animals and clinical subjects. Immediately after trauma acetylcholine levels increased, suggesting enhanced activity of the central nervous system. This was followed by predominant activity of the sympathetic nervous system, as evidenced by increased catecholamine levels. During the later postoperative period, histamine levels also increased. There appears to be a good correlation between increased histamine content and fibroblastic proliferation in the traumatized area. These observations suggest that levels of the neurohumors increase in a definite sequence after trauma, facilitating the process of repair, regeneration, and normal convalescence. Therefore, trauma should be minimized during repair and the patient should receive proper sedation and rest during the pre- and postoperative periods, since each quantum of response depends upon the amount of sensory input that occurs at the time of trauma and in the post-traumatic period.


[Comparative investigations concerning the influence of pilocarpine and aceclidine on the glucose consumption of surviving ocular tissues (author's transl)]. Conservative glaucoma therapy repeated during a whole lifetime raises the question of medicamentuous tolerance. Recently, cultivation of normal lens epithelia and of iris tissue has made possible determination of metabolic functions with and without application of different medicaments. Preceding experiments revealed a diminution in glucose consumption by these tissues after addition of pilocarpine. Comparative studies show 0.25 g/100 ml medium to be maximum concentration of aceclidine allowing cellular survival. With this dilution, the glucose metabolism of lens epithelia was much more sensitive to aceclidine than to pilocarpine. This was less true for iris tissue. Accordingly, the lens of an aceclidine-treated patient requires careful observation.


Plasma lorazepam levels. A study following single dose administration of 2 and 4 mg by different routes. Plasma lorazepam levels were studied following single doses of 2 mg and 4 mg in patients and volunteers. There was a slightly more rapid uptake of drug following deep intramuscular injection than when taken by mouth. Plasma levels declined rapidly following intravenous injection (t1/2 of approximately 2-3 hours). Two hours after giving the drug the plasma levels were similar irrespective of the route of administration. The second phase decline was very slow and one third of the peak concentration was still in the blood at 24 hours.


[Metabolism of glucides and polyols and disorders of their utilization]. The first part of this report is devoted to a description of the metabolic pathways of the principle carbohydrates and polyols. The second part is concerned with the utilisation of these energy-providing substrates, when used in parenteral alimentation. Notions emerge of "total clearance" and of coefficient of utilisation in relation to the rate of administration for each of them. Finally, the third part deals with disturbances of carbolydate and polyol metabolism during stress and diabetes and the possiblilities of the development of hyperlactatemia according to the substances used or the pathological circumstances encountered. A table attempts to summarise the potential complications in comparison with all the substrates used in parenteral alimentation.


[Glucide intolerance and its pathogenic mechanisms during parenteral feeding]. Hyperglycaemia during parenteral alimentation occurs either as a result of an error in the supplies provided or as a result of diminished carbohydrate tolerance. The circumstances surrounding the development of carbohydrate intolerance are essentially : severe infections, major catabolic states, renal insufficiency, extensive burns, pancreatic problems and diabetes. From a pathogenic standpoint, there are two dominant elements : disturbances in hepatic gluconeogenesis and changes in insulin secretion and in resistance to insulin. The physiopathology is dominated by the risk of hyperosmolarity. Hypoglycaemia occurs most frequently as the result of a manit fest error : too sudded interruption of carbohydrate supplies or two high dosage of exogenous insulin.


Neonatal gastric pH.

Neonatal gastric pH. The pH of gastric juice, obtained 3 to 4 minutes after birth in 158 unselected neonates, varied between 7.5 and 8.5 in 8 meconium-containing specimens and ranged from 1.4 to 7.8 in 150 meconium-free samples. In mature infants of the latter group, pH was (1) significantly lower after vaginal delivery than after cesarean section; (2) tended to be lower after section preceded by labor than after elective section; and (3) was lowest after precipitate delivery. In premature infants, pH was above 7 regardless of mode of delivery. Three was no correlation between neonatal gastric pH and pH of simultaneously obtained maternal gastric juice, pH of amniotic fluid, pH of cord blood duration of rupture of membranes, birth weight, or Apgar score. It was concluded that the mature human fetus produces gastric acidity in response to stresses associated with labor and vaginal delivery. The possibility of a low gastric pH and the resultant pulmonary damage if aspirated must be considered in the initial care of thew newborn with poor muscle tone or reflex activity as well as in the anesthetic management of neonates undergoing emergency surgery.


[Circulating lipids in prolonged postoperative parenteral feeding: their changes depending on the nature of energy intake]. Repeated estimations of circulating total lipids and various fractions were made during the postoperative period in patients receiving parenteral nutrition for more than ten days. The patients were paired in relation to the underlying pathology and divided into two group : the first receiving energy supplements in the form of carbohydrates alone, whilst the second received part of this supply in the form of lipid emulsions (Trive 1000).  Total lipids and various plasma fractions, with the exception of free fatty acids, increased progressively during the postoperative period, regardless of the inital values and independently of the lipid content of the nutrition fluids given. There was no significant difference between those patients who received lipid emulsions and those who did not. Free fatty acids remained at levels slightly greater than normal in the group which did not receive lipids. They were significantly higher in the group given lipid emulsions, though it was not possible to precisely define their orgin.


[Fat overload during intake of lipids].

[Fat overload during intake of lipids]. A review of the recent literature indicates that fat overload during the infusion of lipid emulsions remains rare, at least clinically. Nevertheless, two pictures may be seen, the physiopathology of which is discussed:--overload syndrome, with an association essentially of general and digestive signs, related to the accumulation of lipids in a certain number of parenchymatous tissues;--fat embolism. Problem may be seen at any time after administration of the solution is begun. Youth seems to predominate amongst the favourising factors. Prevention is possible if rules of caution concerning dose and the rate of administration are respected, combined with biological surveillance involving at laboratory studies at least once a week.


[Disorders of hemostasis during intake of lipids]. Lipid emulsions based upon soya triglycerides and widely used in parenteral nutrition. Haematological surveillance of the patients, and in particular the study of coagulation, shows slight changes in platelet adhesiveness and a transient hypercoagulability state proportional to the concentration of the lipid emulsion. Nevertheless, these findings must be interpreted with the greatest caution, inasmuch as the laboratory tests involved are technically delicate. The results obtained may be contradictory according to the protocol used. Finally, in patients receiving intensive therapy, there are multiple factors (stress, prolonge;d bedrest, hypercatabolism) which may alter coagulation quite independently of the prescription of lipids.


[Blood coagulation during prolonged postoperative parenteral feeding containing lipids]. Blood coagulation was studied in two groups of fifteen patients of the same type during parenteral nutrition lasting for more then ten days and given during the postoperative period. The first batch of patients, taken as a control group, received parenteral nutrition with carbohydrates and proteins, whilst the second group received lipids in addition. Study of blood coagulation, carried out every three days, involved : thromboelastogen on whole blood and plasma, platelet count, platelet adhesiveness and the estimation of factors II, V, VII and X, fibrinogen, plasminogen and fibrinogen break-down products. In the patients not receiving lipids, the normal alterations seen during the postoperative period occurred : normalisation in plasminogen levels and in F.B.P., and an increase in the number of platelets and in fibrinogen level. The presence of lipids in the nutritional fluids resulted in changes in the same direction, but the increase in platelet count was slighter and fibrinogen level, after a slight increase, returned to initial values on the 20th day of parenteral nutrition. Fibrinogen was the only factor which showed any significant difference between the two groups.


[Cholestatic icterus during parenteral feeding in the newborn ano infants]. The authors observed six cases of cholestatis jaundice occuring during parenteral alimentation in the newborn and the infant. In 4 cases, local factors (duodenal atresia, necrosing enteritis of prematurity) or general factors (mucoviscidosis) probably played an essential role. In two cases, the parenteral alimentation seemed to be a main factor, but in one case with excessive and imbalanced supplies of amino acids. Three patients survived, with regression of the jaundice in 15 days to three weeks following the interruption of parenteral alimentation.


[Prevention of metabolic complications in prolonged parenteral feeding]. Modern technics in parenteral nutrition can prevent metabolic and infectious complications. Supplying a patient with a daily ration according to his needs in the form of either hyper or normo-nutrition serves as the basis for this prevention. All that is required for success is a deep venous approach and regular biological and clinical surveillance of the patient. The authors conclusions are based on a study of 1.800 patients.


[Metabolic disorders in experimental hepatic insufficiency and parenteral feeding]. Anhepatia is a complete experimental model for the study of metabolic disturbances associated with hepatic insufficiency. The authors report their experience based upon 16 cases of total hepatectomy in the dog. The animals received different types of hepatic assistance : group 1 (2 dogs), carbohydrate assistance (glucose 7 g/kg/24 h) ; group II (2 dogs), carbohydrate and haemostatic assistance : homologous plasma 50 ml/kg body weight per 24 hours previously stored at - 80 degrees C ; group III (2 dogs) same nutrition as in group II, combined with amino acids : leucine 53, 3 mg, isoleucine 46, 6 mg ; valine 53,3 mg alpha-ketonic acid (lg/kg/24 h) ; group IV (10 dogs, same nutrition as in group III, combined with supplements in neuro transmitter precursors : 6 dopa 1,66 mg, 5 hydroxy-tryptophane 2 mg and choline 16, 66 mg/kg/24h. Metabolic disturbances and their course were analysed in detail. Results : (mean survival) - group I : 24 +/- 4 hours; group II : 36 +/- 9 hours; group III : 42 +/- 4 hours; group IV : 90 +/- 9 hours. The average survival differed significantly for groups III and IV (p less than 0,01) in comparison with control groups I and II. The replacement of essential hepatic functions such as blood glucose regulation, coagulation, detoxication and the provision of neurotransmitter precursors made possible 4 days survival after total hepatectomy.


[Three cases of coma during parenteral alimentation]. Three cases of coma occuring during parenteral alimentation are presented. The common factor was that they occurred during the phase of realimentation in patients with severely impaired nutrition. In two cases out of three, the coma marked the beginning of the absolute or relative anabolic period. They were studied clinically, biologically and electrically. In one case only, major hypophosphataemia was discovered (2mg/l). Of the three patients, one only died 4 days after the onset of coma, whilst clinical and biological characteristics had returned to normal.


[Deficiencies in trace elements during parenteral alimentation]. During recent years, techniques of parenteral alimentation have seen the development of complications due to deficiencies in trace elements. The main deficiency states described involve iron, copper and zinc. In addition to these three trace elements, manganèse and cobalt are included in this study. The metabolism, physiological role, needs in parenteral alimentation and clinical and metabolic effects of possible deficiency are described for each of these elements.


[Routine comparison of trace element deficiencies during parenteral alimentation]. In 50 patients aged between 3 and 84 years treated in a multidisciplinary Intensive Care Unit and receiving parenteral alimentation, deficiency in certain trace elements or electrolytes (Cu++, Zn++, Mn++, Co++, PO-4, Mg++) was prevented or treated by the administration of a glucose solution (MB 147 G) enriched in trace elements. The aim of the present study was to demonstrate, on the basis of assay of serum levels of the trace elements involved, with the exception of Mn and Co, the effectiveness of treatment. Reference values were determined on the one hand in healthy individuals for normal figures and secondly on subjects included in the study, already on parenteral alimentation for several days, before treatment with MB 147 G, in order to demonstrate the existence of a deficiency (patient control values). In the case of PO--4, however, the patient control values concerned at one and the same time subjects in the study before treatment with MB 147 G and other patients receiving parenteral alimentation who were not part of the trial. MB 147 G solution was presented in units of 500 ml associated with glucose of varying concentrations (15 p. 100, 30 p. 100, 50 p. 100). The average daily amount administered, over a period of 236 days, was 3 unites per 24 hours, corresponding to an intake of copper of 3.78 mg, 3.90 mg of zinc, 0.20 mg of manganese, 0.24 mg of cobalt, 363 mg of magnesium, 240 mg of calcium and 15 mEz of phosphates. The results show that levels of copper, zinc, magnesium and phosphates were low during parenteral alimentation. The administration of MB 147 G resulted in a significant increase in these values, without there being any evidence of accumulation.


[Hydrogen ion metabolism and parenteral alimentation]. The necessity of using solutions and emulsions in parenteral nutrition of solutions directly utilisable by the cell has led to the manufactur of substances with a composition and physico-chemical nature which greatly alters H+ ions balance. Theoretical calculation and various experimental protocois have shown that the production of H+ ions may be multiplied by 3 or 4. This explains the complications of metabolic acidosis seen in association with prolonged parenteral nutrition. There are two main sources : lipid emulsions and amino-acid solutions. One litre of lipid emulsion may provide 37 to 60 mEq of H+ ions. According to their formula, amino acid solutions may consume, or more often provide, large amounts of H+ ions (up to 20 mEq/gram of nitrogen). These physiopathological concepts should be taken into consideration in defining the compositions of fluids for parenteral nutrition, in particular when renal function is impaired.


[Metabolic problems associated with ethanol administration]. After reviewing the effects of ethanol on different metabolic processes, the author defines the doses of this alcohol which may be used in parenteral alimentation, its advantages and disadvantages. The latter in general outweigh the advantages and limit its use in parenteral nutrition.


[Inherited abnormalities in amino acid metabolism]. Inherited abnormalities in amino-acid metabolism, which up to a few years ago remained curiosities in the domain of the specialist, are now becoming almost of everyday significance. Improved knowledge, as a result of the combined efforts of paediatricians, genetecists and biochemists, has led to substantial progress, both clinically (diagnosis and treatment) as well as in the understanding of intermediary metabolisms and enzyme reactions. This "jigsay puzzle", the pieces of which fall gradually into place, has already resulted in spectacular results, the best example of which is that of phenylketonuria. In the past this disease inevitably led to increasing brain damage and has now become a purely biochemical disorder wince early diagnosis and appropriate diet enable the young infant to become a normal child and remain a healthy adult. These data are illustrated here.


[Biological and hormonal balance during oral nutrition, exclusive perenteral and mixed nutrition in children]. Exclusive parenteral nutrition and mixed feeding provided by the infusion of a solution of amino-acids, glucose and trace elements by a drip method, result in changes in biological and hormonal balance which have been compared with balance in the child on normal oral feeding. The 24-hour cortisol cycle tends to be attenuated. Blood glucose and, above all, blood insulin levels show major variations. Plasma free fatty acid levels, stable in exclusive parenteral nutrition, in general vary with mixed nutrition. On the basis of these findings, a constant flow should be ensured by use of a pump during exclusive parenteral nutrition. In mixed feeding, meals should be frequent and energy supplies given in the form of slowly absorbed carbohydrates.


[Clinical signs and etiological aspects of metabolic encephalopathies excluding liver encephalopathy and realimentation syndrome]. The clinical picture of metabolic encephalopathies has no aetiological specificity. It combines disturbances in conscious level dominated by disorientation and time and space and disturbances in motor activity, in particular tremor and asterixis. For each of the aetiologies studied, the following are considered: the circumstances of onset, the clinical and laboratory picture, the physiopathology and the treatment. From a diagnostic standpoint, particular emphasis should be placed upon the circumstances of onset which alone give any indication. The majority of these encephalopathies are caused by a lack of respect for simple rules in parenteral alimentation or by deficiencies. It is thus essentially an iatrogenic pathology. Treatment should be above all preventive.


[Amino acid disturbances in liver failure]. We have attempted to trace the development of the concept which stresses that the encephalopathy often seen with hepatic failure may be the result of derangement in amino acid metabolism in patients with failing livers. We have also proposed that the infusion of solutions which may normalize these plasma amino acids and decrease the amount of toxic aromatic amino acids within the circulation may help in the amelioration of the symptoms of hepatic encephalopathy. We believe that in our studies to date, the use of this solution has been a significant advantage in the care of these extremely ill patients. We would hope that when the solution becomes generally available, the provision of such solutions to such patients will result in the salvage of some lives lost at the present time because of hepatic failure and encephalopathy.


[Comas of realimentation].

[Comas of realimentation]. Parenteral realimentation in the patient with severely impaired nutrition is sometimes associated with the development of a metabolic encephalopathy with coma. The onset of this complication is sudden. The coma is deep, accompanied by signs of neuromuscular hyperexcitability and very marked hyperventilation. It generally regresses rapidly, without sequellae. The onset of such a realimentation coma should be feared in the presence of a certain combination of conditions : severely impaired nutrition, parenteral alimentation with a high level of nitrogen and calories, transfer to the anabolic phase (perfect carbohydrate utilisation, fall in blood phosphate levels with hypophosphaturia, sometimes very marked positivisation of nitrogen balance) and, sometimes, mild premonitory clinical signs. The relationship between this type of complication and hypophosphoraemia is quite definite, but the fall in serum phosphates would not appear to be directly responsible for the coma. The exact mechanism is not known. In order to avoid such complications, great caution should be observed in both the quality as well as the quantity of the intake, as well as in the clinical and laboratory surveillance of the malnourished patient undergoing realimentation.


[Electroencephalographic study of functional metabolic encephalopathies and comas during parenteral alimentation]. This review deals with the common E.E.G. characteristics of dysmetabolic encephalopathies and described the particular features of hepatic, respiratory and renal encephalopathies, as well as those resulting from a disturbance in carbohydrate, water and electrolyte metabolism. These now classical data are compared with the principal electroclinical appearances seen during parenteral alimentation : slow, ample wases, non-reactive, associated with a calm coma; overall depression of basal rhythm, with excessive myogram activity and corresponding to paroxysms of muscular hypertonia seen during the coma.


Effect of aflatoxin on the humoral and cell-mediated immune systems of the chicken. Chickens fed 2.5 microgram of aflatoxin/g of diet from 2 to 4 weeks of age or from hatching to 4 weeks were deficient in cell-mediated immunity, as measured at 4 weeks of age by the graft-versus-host reaction. Delayed-type hypersensitive skin reactions to tuberculin were also reduced in chickens given dietary aflatoxin from hatching to 7 weeks of age. Humoral immunity, as measured by the ability of 4-week-old chicks to produce natural agglutinins to rabbit red blood cells, was not significantly altered by dietary aflatoxin. A significant decrease in concentrations of serum immunoglobulins (Ig) IgG and IgA, but no IgM, however, did occur in chicks given dietary aflatoxin from hatching to 4 weeks or between 2 and 4 weeks of age. Aflatoxin consumption from 0 to 2 weeks of age produced no marked effect on either cell-mediated or humoral immunity in 4-week-old chicks.


Less dialysis-induced morbidity and vascular instability with bicarbonate in dialysate. We devised three protocols to test the postulate that increased morbidity during high-efficiency dialysis with large-surface-area units (LS) might be due in part to the increased flux of bicarbonate out and acetate into the patient inherent is LS dialysis. The first protocol showed that with LS-acetate dialysis there was a marked fall in plasma bicarbonate and Pco2 during the first 3 to 4 h, followed by a rapid rise in bicarbonate above normal and return to control in Pco2. With LS-bicarbonate dialysis, these oscillations were largely eliminated. A second double-blind protocol showed that central nervous system-type symptoms noted during and after LS-acetate dialysis were reduced significantly by switching to LS-bicarbonate dialysis. The third protocol showed that with LS-carbonate the tolerable rate of ultrafiltration could be increased 67% compared with LS-acetate dialysis.


Metabolic acidosis after hyperalimentation with casein hydrolysate. Occurrence in a starved patient. A 29-year-old woman with short bowel syndrome and prolonged starvation developed hyperchloremic metabolic acidosis after initiation of hyoeralimentation with a casein hydrolysate solution. The acidosis was not due to bicarbonate loss but was associated with diminished ability of the kidney to increase urinary acid excretion, particularly titratable acidity. Supplemental parenteral bicarbonate administration was necessary for two weeks until urinary acid excretion rose to normal.


Costs of educating child health associates. Child health associates can be fully prepared to competently diagnose and treat 90% of ambulatory pediatric patients five years after completion of high school at a cost of approximately one-fourth the expense of educating a pediatrician to perform the same tasks and functions. A rational health planning policy should include the extensive utilization of child health associates as primary health care providers for children.


A chemiluminescent effect encountered in the liquid scintillation counting of 32P phosphate. During the course of a kinetic enzyme study of bisphosphoglyceromutase (EC 2.7.5.4.) a chemiluminescent effect was encountered in the liquid scintillation counting of the beta-emission from 32Pi. A method is described for overcoming the effect, thereby permitting the accurate determination of the radioactive compounds. It is important to check for chemiluminescent effects early in the development of new procedures that involve liquid scintillation counting.


The origin of the acidosis in hyperlactataemia. Anaerobic glycolysis produces lactate, ATP, and water but there is no net change in the number of hydrogen ions: it does not produce lactic acid. The acidosis usually associated with hyperlactataemia is caused by hydrolysis of the ATP, with release of hydrogen ions. By contrast, ATP turnover by aerobic mechanisms is not acidifying because the released hydrogen ion is reutilised as more ATP is formed. Gluconeogenesis from lactate does not utilise hydrogen ions directly--in fact, it produces them. The associated net H+ utilisation is caused by the aerobic generation of the ATP and GTP required to drive glycolysis in reverse. It is suggested that only by understanding these important biochemical facts can the clinician found his diagnosis and treatment on a firm, rational basis.


[Study of E. coli penicillin amidase. The pH dependence of the equilibrium constant of ampicillin enzymatic hydrolysis]. The equilibrium parameters of the hydrolysis of ampicillin catalysed by penicillin amidase were determined within the pH range of 4.5 to 5.5. The values of the ionization constants of the carboxy group of D-(-)-ALPHA-AMINOPHENYLACETIC ACID (PK1=1.80) and amino group of 6-aminopenicillanic acid (pK2=4.60) were estimated and pH-dependence of the effective free energy of ampicillin hydrolysis was calculated. It was shown that the thermodynamic optimum of ampicillin synthesis was at 3.20 (the value of the effective free energy under the experimental conditions was 3.27 kcal/mole). The value of the "true", pH-independent free energy of hydrolysis (deltasigma) of the amide bond in the ampicillin molecule was determined to be equal to 9.72 kcal/mole. The thermodynamic parameters of ampicillin and benzylpenicillin hydrolysis were compared. The amino group in the alpha-position of phenylacetic acid was shown to have a significant effect on the values of "true" free energy of hydrolysis of the penicillin amide bond and free ionization energy in the system.


[Use of the penicillanic acid formation reaction for the analysis of penicillins. The quantitative determination of oxacillin]. A procedure for quantitative determination of oxacillin by means of spectrophotometry is described. The procedure is based on the capacity of penicillins to transform on heating in acid buffer solutions containing copper ions into respective penicillenic acids having specific absorption characteristics in the UV-spectrum region. The amount of oxacillin was estimated by the optical density of the solutions at a wave length of 335 nm corresponding to the absorption maximum of the penicillenic acid of oxacillin. The optimal conditions for the reaction of the penicillenic acid formation, i.e. pH of the buffer solution and the level of the copper ions in it, time and temperature of heating were determined. The data on the comparison of the specificity and reproducibility of the new method with the currently used ones for determination of oxacillin are presented.


Stability and blood level determinations of cefaclor, a new oral cephalosporin antibiotic. Cefaclor solutions in pH 2.5 and 4.5 buffers contained at least 90% of their initial activity after 72 h at 4 degrees C. Samples in pH 6.0, 7.0, and 8.0 buffers contained 70, 46, and 34%, respectively, of their initial activity after 72 h at 4 degrees C. After 72 h at 25 degrees C, samples prepared with pH 2.5, 4.5, 6.0, 7.0, and 8.0 buffers contained 95, 69, 16, 5, and 3%, respectively, of their initial activity. After 72 h at 37 degrees C, cefaclor solutions in pH 2.5 buffer contained 80% of the initial activity, whereas samples prepared in pH 4.5, 6.0, 7.0, and 8.0 buffers contained less than 20%. Laboratory-prepared plasma and serum samples showed an 8% loss in activity when incubated for 6 h at 4 degrees C, a 51% loss when incubated for 6 h at 25 degrees C, and a 48% loss when incubated for 2 h at 37 degrees C. Clinical samples demonstrated a similar stability pattern. Degradation rates for cefaclor in commercially prepared serum increased from 4- to 10-fold in comparison to rates obtained when samples were made in human serum freshly prepared in our laboratory. Consequently, serum standards should be made in freshly prepared human serum.


Mal perforans and spina bifida occulta.

Mal perforans and spina bifida occulta. A 26-year-old man had mal perforans pedis as the principal manifestation of spina bifida occulta with an associated intradural lipoma. Two lumbosacral dimples and a tuft of hair overlying the spina bifida led to the diagnosis. We include a review of the causes of mal perforans and a review of spina bifida occulta and associated abnormalities.


Alclofenac and D-penicillamine. Comparative trial in rheumatoid arthritis. Forty-six patients with rheumatoid arthritis, 22 receiving D-penicillamine and 2j alclofenac, took part in a 6-month single-bind external observer trial to compare the efficacy and toxicity of these drugs in the treatment of severe rheumatoid arthritis. Both drugs were active and similar in their efficacy at 6 months as judged by clinical and laboratory measurements. Penicillamine was active therapeutically by 3 months, one month before alclofenac. 9 patients, 8 on alclofenac and one on D-penicillamine, had to stop treatment because of lack of effect or toxic effects. Skin rashes within the first week of treatment were a major problem with alclofenac and led to 6 withdrawals.


Evaluation of physician assistants in rural primary care. The advent of physician assistants raises questions regarding their appropriate utilization and monitoring. To determine practice patterns of physician assistants in settings independent of training programs, we studied 14 primary care practices in the rural southeast. Detailed observations, including appropriateness of medical care, were made on 788 outpatient-provider encounters. Physician assistants handled minor medical problems well and 61% of the patients observed in these practices fitted this category. Three practice patterns were observed: all patients were seen by the assistant initially, followed by the physician; patients managed concurrently by physician and assistant were not preselected; and patients with specific problems were assigned to the assistant. Properly managed, each of these patterns yielded competent care. Using these observations, proposed models of management and audit are presented for each practice pattern.


Allergic angiitis and granulomatosis. Prolonged remission induced by combined prednisone--azathioprine therapy. Two patients with acute, rapidly progressive generalized vasculitis initially had symptoms of asthma. Progressive increase in severity of asthma was followed by systemic disease, including pulmonary infiltrative disease, mononeuritis multiplex, and abdominal pain. Examination of the tissues demonstrated vasculitis with eosinophilia, and clinically both cases appeared in a near terminal state. High-dose prednisone did not induce a remission. In particular, the lesions of mononeuritis multiplex progressed after initiation of high-dose prednisone. The addition of azathioprine to the regimen was followed by a gradual and then complete remission of clinical and laboratory abnormalities, except for some residual nerve damage and asthma of varying severity in the two patients. These two patients, whose cases are classified as the allergic granulomatosis variant of polyarteritis nodosa, have had a remission of seven and almost two years, respectively, after combined prednisone-azathioprine therapy.


[Selective local hyperthermy of tumor tissue. Homogenized energy supply also to deep-seated tissues by high-performance decametric wave coil section plus dual system raster motion (author's transl)]. Based on results of selective tumor tissue hyperacidification and whole-body hyperthermy the paper deals with the advantages of local hyperthermy and gives a review of the various methods of energy supply for hyperthermy in living tissue. It follows a presentation of physical and biophysical fundamentals on selective local hyperthermy of tumor tissue by radiation, by r. f. rotational magnetic fields and by r. f. capacitor fields. Using a gelatine phantom for measuring the energy supply with the various modes of local hyperthermy it is then shown that it is solely the divergent rotational magnetic field which allows to substantially reduce the inhomogeneity of energy supply also in z-direction (increasing depth in body tissue) by introducing a relative raster motion of the applicator in x-y-direction (plane parallel to body surface). It is only the introduction of the x-y raster principle combined with a described dual system array (CMT Selectotherm System) which permits the high and rather homogeneous supply of thermal energy also to deep-seated tumor tissue in the patient without concomitant critical superheating of tissues near the skin. Finally the paper gives a theoretical derivation of the temperature profile for selective local hyperthermy of tumor tissue. With due consideration of heat dissipation by convection and conduction, this endeavour shows rather promising aspects for the therapeutic efficiency obtainable in each case of treatment.


Anxiety, arousal, and autonomic habituation. The interaction of anxiety and autonomic activation as a factor in the development and persistency of pathological anxiety was investigated with the aid of self-rating procedures and a habituation experiment. The state of activation was varied systematically in 40 normal subjects by various experimental conditions and by the administration of a tranquilizer. The degree of anxiety and activation were able to be differentiated in the investigated range of mean attentiveness. Anxious expectancy is perceived in particular as subjective anxiety. Fatigue and sedation, on the other hand, demonstrate subjective and autonomic desactivation. Corresponding differences can be demonstrated for the anxiolytic and sedative effects of tranquilizers. The time course of habituation is a more exact indicator than the amplitude of the orienting response. Cognitively provoked apprehensiveness, thus, appears to be qualitatively different as compared to psychoautonomically caused anxieties of psychiatric disorders.


Purification of human diploid fibroblast interferon by immobilized neuraminidase. An efficient method for the purification of human fibroblast interferon (IF) based on binding via the N-acetyl neuraminic acid (N-ANA) residue of the IF molecules to the immobilized neuraminidase has been developed. Binding of IF occurred at pH 4.5 and elution of the bound activity was effected at pH 9.5. Specific activity of IF in the alkaline eluate was increased by a factor in excess of 200 and the IF thus recovered had probably retained most of the N-ANA moieties.


[State of the microcirculation in periarteritis nodosa based on morphological and biomicroscopic data]. The status of microcirculation in periarteritis nodosa was studied by impregnation method (serous membranes) and biomicroscopy (eyeball conjunctiva). Pathological and adaptative changes in terminal vessels were found. It is suggested that the involvement of microcirculation ways in this disease is systemic and that morphological changes in microvessels are comparable to biomicroscopic changes.


Oxidative phosphorylation. Halide-dependent and halide-independent effects of triorganotin and trioganolead compounds on mitochondrial functions. 1. Each of five triorganotin and five triorganolead compounds was shown to perturb mithochondrial functions in three different ways. One is dependent and two are independent of Cl- in the medium. 2. Structure-activity relationships for the three interactions are described, and compounds suitable as tools for the separate study of each process are defined. 3. In a Cl- -containing medium trimethyltin, triethyltin, trimethyl-lead, triethyl-lead and tri-n-propyl-lead all produce the same maximum rate of ATP hydrolysis and O2 uptake; this rate is much less than that produced by uncoupling agents such as 2,4-dinitrophenol. 4. Increase in ATP hydrolysis and O2 uptake are measures on energy ultilization when triogranotin and triorganolead compounds bring about an exchange of external C1- for intramitochondrial OH- ions. Possible rate-limiting steps in this process are discussed. 5. In a C1- -containing medium ATP synthesis linked to the oxidation of beta-hydroxybutyrate or reduced cytochrone c is less inhibited by triethyltin or triethyl-lead than is ATP synthesis linked to the oxidation of succinate, pyruvate or L-glutamate. 6. The inhibition of ATP synthesis linked to the oxidation of both beta-hydroxybutyrate and reduced cytochrome c consists of two processes: one is a limited uncoupling and is C1- -dependent and the other is a C1- -independent inhibition of the energy-conservation system. 7. The different sensitivities to inhibition by triethyltin of mitochondrial functions involving the oxidation of beta-hydroxybutyrate and succinate are compared and discussed.


Demonstration of the phosphorylation of acetyl-coenzyme A carboxylase within intact rat epididymal fat-cells. Intact rat epididymal fat-cells were incubated with 32Pi and the intracellular proteins separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. One of the phosphorylated proteins has the same RF value as [14C]biotin-labelled acetyl-CoA carboxylase purified from fat-cells and is specifically precipitated after incubation with antiserum raised against acetyl-CoA carboxylase. No significant changes in the extent of phosphorylation of acetyl-CoA carboxylase were detected after exposure of the cells to insulin.


Isolation of proteoglycans from human articular cartilage. Proteoglycans were extracted from normal human articular cartilage of various ages with 4M-guanidinium chloride and were purified and characterized by using preformed linear CsCl density gradients. With advancing age, there was a decrease in high-density proteoglycans of low protein/uronic acid weight ratio and an increase in the proportion of lower-density proteoglycans, richer in keratan sulphate and protein. Proteoglycans of each age were also shown to disaggregate in 4M-guanidinium chloride and at low pH and to reaggregate in the presence of hyaluronic acid and/or low-density fractions. Osteoarthrotic-cartilage extracts had an increased content of higher-density proteoglycans compared with normal cartilage of the same age, and results also suggested that these were not mechanical or enzymic degradation products, but were possibly proteoglycans of an immature nature.


Activation of immobilized acetyl-Coenzyme A carboxylase. Partially purified acetyl-CoA carboxylase was covalently bound to a Sepharose 4B matrix. Although aggregation was thus prevented, the enzymic activity was stimulated by citrate and isocitrate.


Synthesis of the native copper(II)-transport site of human serum albumin and its copper(II)-binding properties. A derivative of the native-sequence tripeptide of the specific Cu(II)-transport site of human serum albumin, L-aspartyl-L-alanyl-L-histidine N-methylamide, was synthesized, and its binding to Cu(II) was examined to determine the influence of the side-chain groups on the Cu(II) binding. The equilibria involved in the Cu(II)-L-aspartyl-L-alanyl-L-histidine N-methylamide system were investigated by analytical potentiometry. Three complex species were found in the pH range 4-10. The same species were identified in both the visible and circular-dichroism spectra. The main species present in the physiological pH range is shown to have the same ligands around the square-planar Cu(II) ion as those reported for albumin and tripeptides diglycyl-L-histidine and its N-methylamide derivative. The results obtained from competition experiments showed that this tripeptide has a higher affinity towards Cu(II) than has albumin itself. The overall findings are compared with those from albumin. At neutral pH the side chains do not play any important role in the Cu(II) binding, but at low pH the beta-carboxyl group of the N-terminal aspartic residue becomes important. A possible competition site on albumin for Cu(II) at low pH is discussed.


The binding of copper ions to copper-free bovine superoxide dismutase. Kinetic aspects. The kinetics of reconstitution of bovine superoxide dismutase from Cu2+ and the copper-free enzyme have been studied by activity, u.v.-absorption, electron-paramagnetic-resonance and pulsed-nuclear-magnetic-resonance measurements. The process appears to be first-order up to 80% completion in most conditions, and is pH-dependent, with an apparent pK of 6.5. U.v.-absorption and solvent proton relaxation rate measurements show that fast binding of Cu2+ occurs, and the initial ligands are likely to be, at least in part, those of the native active site. The recovery of the native activity and spectroscopic properties is a slow process with activation energies of 92 kJ/mol at pH 5.3 and 8.4kJ/mol at pH 8.1 and can be described as a rearrangement of the site around the bound metal. The rate of this process is lower in partially recombined protein samples, probably because of intersubunit interactions.


Partial purification and characterization of a pyruvate dehydrogenase-complex-inactivating enzyme from rat liver. An enzyme inactivating the pyruvate dehydrogenase complex (inactivase) was purified about 8000-fold from rat liver by differential centrifugation, acid extraction of a lysosomerich 25000 g pellet, acetone fractionation, and adsorption on calcium phosphate gel. By exclusion chromatography on Sephadex G-100 a molecular weight of 21 000 was estimated. The purified enzyme was most stable at pH 5.8 in potassium phosphate buffer, and at pH 4.5 in McIlvaine buffer. At high dilutions the enzyme was very labile and was remarkably stabilized by high salt concentrations. Enzyme activity is inhibited by native rat blood serum, iodoacetamide and leupeptin, but not by phenylmethanesulphonyl fluoride, suggesting that it belongs to the class of thiol proteinases. Among various enzymes tested, only 2-oxoglutarate dehydrogenase was attacked by the inactivase to a similar extent to the pyruvate dehydrogenase complex. Studies on the inactivation mechanism indicate that although the overall reaction is completely lost after treatment with inactivase, each individual step of the multienzyme complex retains full catalytic activity. As judged from sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the transacetylase subunit appears to be degraded into several smaller fractions.


Fatty acid synthesis in the regenerating liver of the rat. 1. Synthesis de novo of fatty acids in the rat liver, measured per g wet wt. of tissue, was increased by a factor of about two, between 1 and 4 days after partial hepatectomy, compared with rates in sham-operated control rat livers. 2. There were no associated changes in the rates of liver cholesterol synthesis or of adipose-tissue fatty acid synthesis in rats after partial hepatectomy, compared with rates in sham-operated rats. 3. In regenerating livers, perfused under three different conditions, there was no alteration in the capacity for fatty acid synthesis compared with that of control rats. 4. The increased synthesis of fatty acids in regenerating liver was associated with insignificant increases in plasma concentrations of tricylglycerols and free fatty acids, with a decrease in content of liver glycogen, and with no change in hepatic activity of acetyl-CoA carboxylase. 5. The accelerated rate of synthesis of fatty adids in regenerating liver appears not to be due to any intrinsic alteration in hepatic capacity for fatty acid synthesis, but it may be caused by the continuous action on liver of unidentified circulating factors.


The extraction of antiepileptic drugs. Model study on partition coefficients. To compare the extraction behaviour of hexane, toluene, chloroform, 1,2-dichloroethane, diethylether, ethyl acetate and acetone towards phenobarbital, phenytoin, primidone and carbamazepine, partition coefficients of these drugs between the above solvents and water (saturated with ammonium sulphate when working with acetone) were measured at different pH-conditions. Acetone rendered the highest partition coefficients for all drugs. Ethyl acetate was second in the range, except for carbamazepine, which showed a specific affinity for the chlorinated hydrocarbons.


Intracellular localization of hepatic propionyl-CoA carboxylase and methylmalonyl-CoA mutase in humans and normal and vitamin B12 deficient rats. The intracellular localization of the enzymes in the vitamin B12 dependent pathway which involves the oxidation of propionate was studied in rat liver obtained from normal and vitamin B12 deficient rats as well as from man. The subcellular site of propionyl CoA carboxylase and the vitamin B12 dependent-methylmalonyl CoA mutase were determined. All of the activity of these two enzymes was demonstrated to be in the mitochondria and those enzymes were shown to be loosely bound to the inner membrane-matrix portion of the mitochondria. Vitamin B12 deficiency did not alter the subcellular localization. Finally, a rapid enzymatic assay for methylmalonyl-CoA mutase was described.


Metabolic effects of beta-sympathomimetic drugs and dexamethasone in normal and diabetic pregnancy. Marked hyperglycaemia and hyperinsulinaemia were observed in two normal women in premature labour treated with an intravenous infusion of a beta-sympathomimetic drug and intramuscular dexamethasone injections. Similar therapy in a chemical diabetic patient caused diabetic ketoacidosis, while treatment of an insulin dependent diabetic with dexamethasone alone resulted in a major increase in her insulin requirements. It is suggested that the diabetogenic effects of beta-sympathomimetic drugs and dexamethasone may be additive and that regular plasma glucose estimations should be made when they are used, especially in patients with impaired glucose tolerance.


Renal tubular function in normal pregnancy. Maternal plasma and urine concentrations of beta2- microglobulin and gamma-glutamyl transpeptidase (EC:2.3.2.2) were measured serially in normal pregnancy. These parameters remained constant throughout and can therefore be used to evaluate tubular function in pregnancies complicated by renal disorders.


Electron spin resonance and nuclear relaxation studies on spin-labeled glutamate dehydrogenase. The reaction of glutamate dehydrogenase with two different stable nitroxides (spin labels) is reported. The two compounds contain a carbonyl and an iodoacetamide group as their reactive parts. The carbonyl compound inactivates the enzyme by the formation of a 1:1 covalent complex after NaBH4 reduction of an intermediate Schiff's base. Evidence indicates that the enzyme is modified at lysine-126 in the active site. The electron spin resonance (ESR) spectrum of spin-labeled enzyme indicates a high degree of immobilization of the nitroxide. The binding of reduced coenzyme NADPH is reflected by a change (immobilization) of the ESR spectrum. Nuclear relaxation of bound substrate, oxidized coenzyme, and inhibitor by the paramagnetic group is observed. This shows the existence of a binding site for these compounds close to the active site. The distances of selected protons of the binding ligands to the nitroxide are calculated. The iodoacetamide spin label reacts with several groups, one of which is not a sulfhydryl. The reaction of this particular group causes inactivation of the enzyme. Protection against this inactivation could be achieved with certain ligands. Only enzyme that was spin labeled without such protection caused paramagnetic relaxation of bound substrate and coenzyme.


Raman spectra of heme a, cytochrome oxidase-ligand complexes, and alkaline denatured oxidase. We report 441.6 nm excitation resonance Raman spectra of oxidized and reduced monomeric heme a-imidazole, cytochrome oxidase-exogenous ligand complexes in various redox states, and alkaline denatured oxidase. These data show that, in reduced oxidase, the cytochrome a3 Raman spectrum has bands at 215, 364, 1230, and 1670 cm-1 not observed in the cytochrome a spectrum. The appearance of these bands in the reduced cytochrome a3 spectrum is due to interactions between the heme a of cytochrome a3 and its protein environment and not to intrinsic properties of heme a. These interactions are pH sensitive and strongly influence the vibrational spectra of both heme a groups. We assign the 1670-cm-1 band to the heme a formyl substituent and propose that the intensity of the 1670 cm-1 is high for reduced cytochrome a3 because the C==O lies in the porphyrin plane and is very weak for oxidized and reduced cytochrome a, oxidized cytochrome a3, and oxidized and reduced heme a-imidazole because the C==O lies out of the plane. We suggest that movement of the C==O in and out of the plane explains the ligand induced spectral shift in the optical absorption spectrum of reduced cytochrome a3. Finally, we confirm the observation of Adar & Yonetani (private communication) that, under laser illumination, resting oxidase is photoreactive.


Use of immobilized light-harvesting chlorophyll a/b protein to study the stoichiometry of its self-association. D. J. Davis & E. L. Gross (1976) Biochim. Biophys. Acta 449, 554-564 previously observed that the light-harvesting chlorophyll a/b protein or chlorophyll protein complex II self-associated as determined by ultracentrifugation. We have determined the stoichiometry of complex formation by immobilizing the monomer on ethylenediamine-Sepharose 4B and determing the ability of immobilized protein to bind the free protein. The amount of soluble protein bound to the immobilized protein increased as the concentration of soluble protein increased. The binding was maximal between pH 7 and 8. The maximum binding was three molecules bound per one molecule of protein immobilized. These results indicate that a tetramer is the intrinsic structural unit of the light-harvesting chlorophyll a/b protein in the chloroplast membrane. Upon complex formation, the chlorophyll fluorescence was decreased without any spectral change. The maximum binding was approximately doubled upon addition of 0.5 mM CaCl2 whereas 5 mM NaCl had no effect. Addition of CaCl2 had no effect on the fluorescence of the monomer. The light-harvesting chlorophyll a/b protein can be isolated from a sodium lauryl sulfate extract of chloroplasts by affinity  chromatography using the immobilized light-harvesting chlorophyll a/b protein.


Kinetic analysis of the acid-alkaline conversion of horseradish peroxidases. The nature of the acid-alkaline conversion of horseradish peroxidases was studied by measuring four rate constants in reactions, E + H+ (k1) in equilibrium (k2) EH+ and E + H2O (k3) in equilibrium (k4) EH+ + OH-, where EH+ and E denote the acid and alkaline forms of the enzymes. The values of k1, (k2 + k3), and k4 were obtained by measuring the relaxation rates of the acid leads to alkaline and alkaline leads to acid conversions by means of th pH jump method with a stopped-flow apparatus. The value of k3 could also be obtained by measuring the rate of reactions between hydrogen peroxide and peroxidases at alkaline pH. The measurements were conducted with four peroxidases having different pKa values: peroxidase A )pKa = 9.3), peroxidase C (pKa = 11.1), diacetyldeuteroperoxidase A (pKa = 7.7), and diacetyldeuteroperoxidase C (pKa = 9.1). The value of k1 was about 10(10) M-1 s-1 in the reaction of the four enzymes while k4 was quite different between the enzymes. The pKa was determined by k3 and k4 for the natural peroxidases and by k1 and k2 for the diacetyldeuteroperoxidases. The mechanism of the acid-alkaline conversion was discussed in comparison with that of metmyoglobin.


The mechanism of oxidation of reduced nicotinamide dinucleotide phosphate by submitochondrial particles from beef heart. 1. Oxidation of NADPH by various acceptors catalyzed by submitochondrial particles and a partially purified NADH dehydrogenase from beef heart was investigated. Submitochondrial particles devoid of nicotinamide nucleotide transhydrogenase activity catalyze an oxidation of NADPH by oxygen. The partially purified NADH dehydrogenase prepared from these particles catalyzes an oxidation of NADPH by acetylpyridine-NAD. In both cases the rates of oxidation are about two orders of magnitude lower than those obtained with NADH as electron donor. 2. The kinetic characteristics of the NADPH oxidase reaction and reduction of acetylpyridine-NAD by NADPH are similar with regard to pH dependences and affinities for NADPH, indicating that both reactions involve the same binding site for NADPH. The binding of NADPH to this site appears to be rate limiting for the overall reactions. 3. At redox equilibrium NADPH and NADH reduce FMN and iron-sulphur center 1 of NADH dehydrogenase to the same extents. The rate of reduction of FMN by NADPH is at least two orders of magnitude lower than with NADH. 4. It is concluded that NADPH is a substrate of NADH dehydrogenase and that the nicotinamide nucleotide is oxidized by submitochondrial particles via the NADH--binding site of the enzyme.


The mechanism of the control of carbon fixation by the pH in the chloroplast stroma. Studies with nitrite-mediated proton transfer across the envelope. 1. CO2 fixation of intact spinach chloroplasts is inhibited by nitrite in a pH-dependent mode. At pH 7.3 in the medium 1 mM NaNO2 and at pH 7.9 5 mM NaNO2 were required for 50% inhibition. 2. The addition of nitrite leads to an acidificiation in the stroma. It appears that nitrite renders the envelope permeable for protons resulting in a breakdown of the pH gradient between the external space and the stroma. 3. In view of earlier results on the pH sensitivity of C02 fixation it is concluded that this pH shift in the stroma is responsible for the observed inhibition of CO2 fixation by nitrite. 4. Octanoate and to some extent also high concentrations of bicarbonate and acetate have a similar effect as nitrite in inhibiting CO2 fixation through an acidification in the stroma. 5. The levels of the intermediates of the CO2 fixation cycle were measured. A strong rise of the levels of fructose- and sedoheptulose biphosphates and a concomitant decrease of the corresponding monophosphates was observed during inhibition of CO2 fixation. It appears that the enzymatic steps of the CO2 fixation cycle responsible for the overall inhibition of CO2 fixation caused by lowering of the H+ concentration in the stroma are fructose- and sedopheptulose bisphosphatase. These two enzymes have an important function in the light regulation of CO2 fixation.


Dicarboxylate transport across the inner membrane of the chloroplast envelope. The uptake of radioactively labeled dicarboxylates into the sorbitol-impermeable 3H2O space (the space surrounded by the inner envelope membrane) of spinach chloroplasts has been studied by means of silicone layer filtering centrifugation. 1. Malate, aspartate and a number of other dicarboxylates are rapidly transported across the envelope leading to an accumulation in the chloroplasts. This uptake proceeds mainly by a counterexchange with the dicarboxylates present there. 2. The dicarboxylate transport shows saturation characteristics allowing the determination of Km and V. 3. All dicarboxylates transported act as competitive inhibitors of the transport. 4. The activation energy of the transport as determined from the temperature dependency is evaluated to be 7 kcal/mol. 5. The rate of dicarboxylate transport is influenced by illumination, the countertransported molecules and the pH in the medium. These changes effect the transport velocity, whereas the corresponding Km values are not altered. 6. It is discussed whether there is more than one carrier involved in dicarboxylate transport in spinach chloroplasts.