[Compared survival of "Escherichia coli" and "Salmonella typhi-murium" in cold water (author's transl)]. The survival rates of Escherichia coli and Salmonella typhi-murium in water are studied at 4, 10 and 20 degrees C and pH 6 and 8, either separately or in mixed culture at four different ratios. S. typhi-murium's survival rate is enhanced at pH 6 and low water temperature. The value of traditional microbiological indicators in assessed in cold water conditions.


[Interferon induced in the hamster by subunits of "Nocardia": essay of protection against rabies (author's transl)]. Lipid-free cells of Nocardia opaca and a cell extract (NWSM) are interferon inducers in syrian hamsters: peak interferon appears 2 h after injection as compared to 8 h in the case of interferon induced by inactivated NDV virus. The interferon induced by Nocardia product is inactivated by a treatment for 30 min at 56 degrees C or 24 h at pH 2: NDV-induced interferon is more stable under these conditions. Hamsters inoculated with NWSM before and after infection with rabies virus are partially protected: their survival time is slightly increased.


Pathogenic free-living amoebae in swimming pools: survey in Belgium. During a short term investigation, 1 outdoor and 15 indoor swimming pools were examined for the presence of amoebae. Of all isolates from 13 pools, 43.6% belonged to the genus Acanthamoeba. The genus Naegleria accounted for 7.3% of the isolates, but none belonged to the pathogenic N. fowleri. A high percentage (70%) of the Acanthamoeba strains tested showed virulence for mice.


Familial fatal Parkinsonism with alveolar hypoventilation and mental depression. The clinical, pathological, and neurochemical characteristics of a newly recognized inherited neurological disorder are reported. Lethargy and mental depression are early symptoms, followed by mild parkinsonism and progressive weight loss. Failure of automatic respiratory control develops and may result in sudden death. Advanced degeneration of the substantia nigra, cell loss and gliosis of the basal ganglia, and focal gliosis in the medulla are seen on pathological study. Degeneration of the nigrostriatal dopaminergic system is evidenced by low levels of tyrosine hydroxylase, dopamine, homovanillic acid, and L-dopa decarboxylase in postmortem brain samples. Taurine concentrations in fasting plasma and CSF are somewhat depressed; brain contents of taurine are within normal limits.


Rosaramicin versus penicillin G in experimental Pneumococcal meningitis. Rosaramicin, a new macrolide antibiotic, was compared with penicillin G in the treatment of pneumococcal meningitis in rabbits. Animals were infected intracisternally with 10(4) colony-forming units of Streptococcus pneumoniae type III (rosaramicin minimal inhibitory/bactericidal concentrations, 0.25/0.5 mug/ml; penicillin G minimal inhibitory/bactericidal concentrations, 0.03/0.06 mug/ml). Treatment was instituted 96 h later. Infusion of rosaramicin at 25 mg/kg per h intravenously for 8 h produced a peak cerebrospinal fluid (CSF) drug concentration of 1.54 mug/ml (range, 0.87-3.6 mug/ml). During this infusion the numbers of pneumococci in CSF decreased from 6.2 +/- 0.5 to 3.36 +/- 1.12 log(10) colony-forming units per ml. Penicillin G, infused at 30 mg/kg per h for 8 h, reached a similar concentration in CSF but caused a greater reduction (P < 0.01) in CSF bacteria, from 6.4 +/- 0.36 to 1.3 +/- 0.67 log(10) colony-forming units per ml. Penicillin G, at 100 mg/kg per day intramuscularly for 5 days, cured 7 of 10 rabbits with pneumococcal meningitis. A higher dose, 300 mg/kg per day for 5 days, was no more efficacious: 11 of 14 rabbits were cured. Rosaramicin at 100 mg/kg per day intramuscularly for 5 days cured only 5 of 15 rabbits with meningitis, but a higher dosage regimen of that drug (250 mg/kg per day intramuscularly) produced acute, fulminant enterocecitis and death within 48 h in seven of eight rabbits. No cytotoxin was detected in the feces of one rabbit with acute enterocecitis. Thus the efficacy of rosaramicin in experimental pneumococcal meningitis, measured by bacterial clearance from CSF and by treatment outcome, was less than that of penicillin G. In addition, high-dose parenteral rosaramicin caused acute, fulminant enterocecitis in a high proportion of treated rabbits.


Viral aggregation: buffer effects in the aggregation of poliovirus and reovirus at low and high pH. The effects of the buffer employed in maintaining a given pH value were tested on the aggregation of two viruses, poliovirus and reovirus. Poliovirus was found to aggregate at pH values of 6 and below, but not at pH 7 or above, except in borate buffer. Reovirus aggregated at pH 4 and below, but was found to aggregate only in acetate or tris(hydroxymethyl)aminomethane-citrate buffers at pH 5. Other buffers tested for aggregation of reovirus at pH 5 (succinate, citrate, and phosphate-citrate) induced little aggregation. No significant aggregation was found for reovirus at pH 6 and above. For both viruses, the most effective aggregation was induced by buffers having a substantial monovalently charged anionic component, such as acetate at pH 5 and 6 or citrate at pH 3. Cationic buffers at low pH, such as glycine, were generally weaker in aggregating ability than anionic buffers at the same pH. These results, when correlated with the isoelectric point of the viruses (poliovirus at pH 8.2; reovirus at pH 3.9) indicated that both viruses aggregated strongly when their overall charge was positive, but only under certain circumstances when their overall charge was negative. Although reovirus aggregated massively at its isoelectric point, poliovirus remained dispersed at its isoelectric point. The conclusion can be drawn that those pH and buffer conditions which induced aggregation of one virus do not necessarily induce it in another.


Abiotic factors affecting the toxicity of lead to fungi. The toxicity of lead (Pb) to fungi in pure culture was influenced by several abiotic factors: pH, inorganic anions, clay minerals, and particulate (humic acid) and soluble organic matter. The toxicity of Pb was potentiated under acidic conditions (pH 5 and 6), and phosphate or carbonate anions reduced the toxicity, apparently as a result of the formation of sparingly soluble Pb salts. Clay minerals (montmorillonite greater than attapulgite greater than kaolinite) and particulate humic acid protected against the toxicity of Pb, presumably as the result of sorption, by cation exchange of the Pb to the exchange complexes, which reduced its availability for uptake by the fungi. Soluble organics, such as tryptone, yeast extract, cysteine, succinic acid, and increasing concentrations of neopeptone, also reduced the toxicity of Pb.


Effects of copper on the sabellid polychaete, Eudistylia vancouveri: I. Concentration limits for copper accumulation. Three experiments with a sabellid polychaete (Eudistylia vancouveri) show the threshold concentration for increasing copper accumulation with time to lie between 3 and 6 micrograms/L total copper in seawater during winter conditions. The branchial crown, probably the major absorptive site, concentrated more copper than the body. Accumulation was influenced by size but not by sex. Our studies indicate that the body burden of copper will increase above natural levels in areas of industrial discharge where copper levels are above the threshold limit for accumulation.


Isolation of an enzyme from soil that degrades the organophosphorus insecticide, crotoxyphos. An enzyme which catalyzed the hydrolysis of crotoxyphos ((E)-1-phenylethyl 3-[(dimethoxyphosphinyl)oxy]-2-butenoate) was isolated from nonsterile and radiation-sterilized Chehalis clay loam with 1.5M Tris (2-(hydroxymethyl)-2-nitro-1,3-propanediol) and partially purified with lead acetate treatment. Two soil-g equivalents of lead acetate purified enzyme in pH 8 buffer hydrolyzed 0.13 mumol of substrate to dimethyl phosphate and alpha-methylbenzyl 3-hydroxycrotonate in 16 hr at 37 degrees C. The enzyme exhibited maximal activity around pH 8.0 and was irreversibly inactivated below pH 5.0 or above pH 10.0. The Km value for crotoxyphos was calculated to be 4.63 x 10(-3) M. The enzyme was stable at 60 degrees C for 10 min, retained activity indefinitely at -10 degrees C, and was completely inactivated within a week at room temperature. When applied to autoclayed Chehalis clay loam, purified enzyme lost 75% of its activity after one week and the remainder within two weeks.


Ultrastructure of the tubular wall of the prepuberal cryptorchid human testis. The tubular wall in the human prepuberal testis of cryptorchid patients has been examined by ultrastructural and histochemical analysis. When compared with the normolocated testes, a disorganization of the tubular wall due to cryptorchidism is apparent. Alterations regarding single elements of the tubular wall are described and a positive correlation seems to exist between the age and the amount of the damage. A possible relation between morphological changes and hormone therapy is discussed.


Serologic diagnosis of gonococcal infection. Infection with N. gonorrhoeae stimulates the production of antibodies to many common, species-specific, and type-specific antigens. The L-antigen is an envelop antigen and antibodies to it could be demonstrated by various methods in more than 90% of the patients after the first 10 days of infection. Serologic tests are not yet available in the United States. If and when they become available, they may be recommended for: (1) Screening asymptomatic men and women, (2) Use as an adjunct diagnostic tool in cases of prostatitis, arthritis, disseminated gonococcal infection, and pelvic inflammatory disease, (3) Use (alone or in conjunction with culture) when specimens must be mailed to a central laboratory, when mailing conditions do not allow for incubation within 24-48 hr, or when proper media and qualified bacteriologists are not available.


[Renovascular hypertension and beta blockers. Theoretical and practical implications]. Beta blockade was instituted in 10 patients with renovascular hypertension due to renal artery stenosis or thrombosis. The treatment was very effective in unilateral stenosis with a normal contralateral kidney (2 kidney Goldblatt) and in fibromuscular dystrophy of the renal artery. On the other hand many failures were observed in hypertension with a single kidney (1 kidney Goldblatt) and in renovascular hypertension with complex lesions or associated renal failure. Although a clear relationship was often observed between the increased plasma renin activity and the antihypertensive effect of beta blockade, this association was sometimes completely erroneous. Beta blockade, which is easy to perform, should be tried out systematically in renovascular hypertension, but, when no result is observed, this therapeutic test should not exclude surgical management thereafter.


[Effect of stunning on pork muscle quality]. The following methods of insensitisation were tested for their effects on muscle pork quality: Electrocoma, 75 V, 15 sec; Electrocoma following intensive electrical stimulation and irritation of various parts of the animals's body, using the insensitising forceps; Dart gun insensitisation. Also tested was the action of muscle relaxants on pork quality. Minor deviation from proper practice of electrical insensitisation was found to cause no significant deterioration in cutlet muscle and ham muscle quality. However, intensive irritation immediately before insensitisation caused higher percentages of PSE (16.7 per cent) and DFD (16.7 per cent). Dart gun insensitisation was quite often followed by poorer quality in the form of watery and bright pork (25.0 per cent). This had detrimental consequences for boiled ham processing (higher loss rates). Sensorial assessment of boiled ham obtained from animals in various categories did not reveal any significant discrepancy between the above techniques. When muscle relaxants were used, muscle pork quality of pigs relaxed by means of succicholine dichloride was much better than that in animals to which electrical insensitisation had been applied. Characteristics of such better quality included slower decline in pH over 45 minutes after death and lower drip loss.


Effect of hexachlorobenzene on enzymes of the steroid metabolism in rat liver. Adult female Wistar rats were fed with a diet containing 0.05% hexachlorobenzene. On the 60th day of this treatment the specific activities of NADPH: delta 4-3-oxosteroid-5 alpha-reductase and the 3-hydroxysteroid dehydrogenases in rat liver microsomes were diminished compared to control rats. The cytoplasmatic 5 beta-reduction was higher in HCB treated rats than in control rats. These alterations of the steroid metabolism lead to increased formation of 5 beta-H-steroids which are known to be inducers of the porphyrin biosynthesis.


[Acute respiratory insufficiency associated with the ingestion of central nervous system depressants]. The study of twenty patients with poisoning by central nervous system depressant drugs is reported. The authors emphasized clinical aspects, complications and mainly the lung function impairment. Barbiturates were the most common drugs, at least in 60% of the cases, used alone or in combination with sedatives and tranquilizers. In 55% of patients the amount of the drug ingested could not be measured. In 60% of the cases we didn't know the period of time between drug ingestion by the patients and their admission to the hospital. On admisson all patients were in coma. The coma was of varying degrees and the deeper the coma the worse was lung function and complications were more frequent. Whenever a cardiovascular collapse was present there was also a high mortality rate. The authors emphasized the importance of a follow-up of these patients in intensive care units, mainly with cardiovascular and ventilatory support.


A model enzymic extracorporeal detoxification system. Preliminary studies at the University of Oklahoma have incorporated the use of a continuous, seal-less blood centrifuge as an extracorporeal detoxification unit to aid in the removal of foreign chemicals from the blood. Detoxification is performed by immobilized enzymes in conjunction with a cofactor (NADPH) bound to a water-soluble macromolecule. A drug enters the device with the plasma and then passes across a semipermeable membrane which serves to retain the cofactor. At this point, a combination of the drug, the cofactor and the enzyme react to form the drug-oxide. The oxide then passes back through the membrane into the blood and back into the body. Concurrently, the macro-NADP+ is reduced by G-6-P and G-6-PD in the cofactor regeneration portion of the device. To facilitate detoxification, the centrifuge is employed to provide plasma rich in toxins, but void of potentially interfering blood components such as platelets and whole blood cells. These components tend to dilute the toxins or adhere to the interfacing membrane, decreasing the permeability of these toxins into the detoxification unit. It is felt that the centrifuge-detoxification combination will provide a potentially efficient hepatic assist device.


The endocrine cells of the rectum of adult ox. In order to identify the endocrine cell types in various parts of the Ruminant gut, we have applied ultrastructural, both morphological and cytochemical, techniques, in parallel to the histochemical ones, to study the rectal mucosa of the adult Ox. In these studies we show that: "EC" cells, of the intestinal type, contain predominantly pleiomorphic granules, which are very electron dense and heavily reactive to "Masson" and "Grimelius" methods; "L" cells are recognizable by their numerous granules, which are fairly homogeneous in shape and osmiophilia. They do not react with "Masson" and are weak or negative to Grimelius's reaction. These granules occur near to others that are less dense, unreactive to "Masson", and that contain an argyrophilic matrix, with an eccentric electron dense core, which does not react with silver; "F-like" cells contain granules which are variable in shape, size and osmiophilia. They are unreactive to "Masson" and weak or unreactive to Grimelius silver; "H" cells contain few, small and uniformly osmiophilic granules. These are unreactive to "Masson" and uniformly reactive to "Grimelius". Our data suggest that the morphology, frequency and distribution of the cell types we have identified in the mucosa of the bovine rectum correspond with those reported in large intestine and rectum of Monogastrics, as by other authors described.


Histochemical studies of some hepatic enzyme activities in rat during extrahepatic common bile duct obstruction. Studies on some enzyme activities as measured histochemically in the ligated common bile duct rat liver are reported. They show principally: an increased GGT reaction in the neoductular cells; a diffusion of the LAP reaction to the neoductuli; an immediate increase and diffusion of the alkaline phosphatase reaction; an increase of G-6-PA and AMPA reactions; an increase, during the first week, of the Phyla activity and a contemporary decrease of the UDPG-GS reaction. No significant changes have been observed in other enzyme activities. By means of some dehydrogenase reactions, morphological evidence that the hepatocytes may be involved in neoductular cells formation was obtained.


Polymorphonuclear leukocyte-inhibitory factor of Bordetella pertussis. II. Localization in the outer membrane. The outer and inner membranes and cytoplasm of spheroplasts of a strain of phase I B. pertussis were fractionated by density gradient centrifugation. The high density vesicles of the outer membranes isolated had the "Pili" characteristic of the bacteria and the same antigenicty as the bacterial surface. Activities for inhibition of polymorphonuclear leukocytes were also almost exclusively localized in this outer membrane fraction. The histamine-sensitizing activity was more dispersed, but its specific activity was also highest in the outer membrane fraction. These results suggest that molecules carrying these activities, which are probably different entities together with the tissue-adhesive pili, form a virulence complex on the surface of phase I organisms of B. pertussis.


Etiological relation between Korean hemorrhagic fever and epidemic hemorrhagic fever in Japan. The first case of epidemic hemorrhagic fever in Japan was seen in Osaka in 1960. The etiologic agent of this disease has not yet been isolated, but a close etiologic relation between Korean hemorrhagic fever and epidemic hemorrhagic fever in Japan has been suspected because of similarities in the clinical and pathological pictures of the two diseases. This relation has now been confirmed serologically by demonstrating specific immunofluorescent antibodies to Korean hemorrhagic fever virus in 19 of 20 sera obtained from subjects 7 to 17 years after an acute attack of epidemic hemorrhagic fever.


Further studies on 5'-Nucleotidase from serum of liver cirrhotic individuals. The kinetic properties of 5'-Nucleotidase  were investigated in untreated patients with liver cirrhosis at 37 degrees C. Mg+2 and Mn+2 were found to activate both normal and liver cirrhotic 5'-Nucleotidase, but Nickel inhibited the enzyme in both systems competitively. Both ATP and adenosine act as inhibitors to 5'-Nucleotidase. The inhibitory constant for ATP was different in normal and liver cirrhotic individuals, 0.1 +/- 0.03 for normal and 0.225 +/- 0.02 for liver cirrhosis. In our investigation, ATP was found to be a competitive inhibitor of 5'-Nucleotidase which compete the substrate (A-5'-MP) for the active site. Inhibition of 5'-Nucleotidase by adenosine is of non-competitive type, for both normal and liver cirrhotic sera. It was observed that both serum 5'-Nucleotidase exhibited pH dependent characteristics; in that there was an optimum substrate concentration at each pH value and the plot of pKm versus pH shows great dependency of km on pH.


The sodium channel in non-impulsive cells. Interaction with specific neurotoxins. The cell line C9 used in this paper has a resting potential of --50 mV (+/- 10 mV) but is unable to generate an action potential upon electrical stimulation. The cell membrane has receptors for the selectivity filter toxin tetrodotoxin as well as for the gating system toxins, veratridine, scorpion toxin and sea anemone toxin. The Na+ channel which remains silent to electrical stimulation in the absence of toxins can be chemically activated by the gating system toxins. This has been demonstarted by electrophysiological techniques and by 22Na+ flux studies. The electrophysiological approach has shown that the sea anemone toxin is able to induce a spontaneous slow-wave activity inhibited by tetrodotoxin. 22Na+ influx analyses have shown that veratridine and the sea anemone toxin produce an important increase of the initial rate of 22Na+ influx into the C9 cell. The stimulation of 22Na+ entry by these gating system toxins is similar to that found using spiking neuroblastoma cells. Veratridine and the sea anemone toxin on one hand as well as veratridine and the scorpion toxin on the other hand are synergistic in their action to stabilize an open and highly permeable form of the sodium channel. Stimulation of 22Na+ entry into the cell through the sodium channel maintained open by the gating system neurotoxins is completely suppressed by tetrodotoxin.


Multiple conformations of deoxyribonuclease A. Their separation at alkaline pH and low ionic strength in the presence of Ca2+. Gel filtration on Sephadex G-100 at pH 9.0 in 1 mM Tris buffer produces denaturation and inactivation of pancreatic DNAase A. Limiting concentrations of Ca2+ in the suspension and elution buffer, reactivates some of the enzyme molecules in an amount proportional to the calcium added. Stable active and inactive forms were separated on Sephadex columns. A model for the conformational role of Ca2+ on DNAase A demonstrates that at least one Ca2+ is involved (Kapp = 8.3 . 10(-5) M) in the correct folding of the polypeptide chain. Na+ was unable to reactivate the enzyme.


Structural studies of a branchiopod crustacean (Lepidurus bilobatus) extracellular hemoglobin. Evidence for oxygen-binding domains. The extracellular hemoglobin of the notostracan branchiopod Lepidurus bilobatus has an apparent molecular weight of 680,000 and may exist in a dissociation-association equilibrium dependent on pH and ligand state. The pigment contains one heme per 18,000 g protein. However, attempts to dissociate the hemoglobin by harsh denaturing conditions results in a 33-34,000 molecular weight polypeptide chain as well as traces of some 62-64,000 molecular weight material. Limited proteolysis of this hemoglobin with subtilisin produces 14,800 and 16,500 dalton heme-containing polypeptides (domains) which bind oxygen reversibly. These domains, isolated by column chromatography, have a heme content similar to the intact pigment. It is proposed that the  intact 34,000 dalton subunit of Lepidurus hemoglobin consists of two linearly linked oxygen binding domains. Oxygen binding properties of the intact hemoglobin show a low oxygen affinity with a slight Bohr effect. In contrast, the isolated domains display a relatively high oxygen affinity and lack a Bohr effect between  pH 7.0 and 8.0. It is apparent that the intact 34,000 dalton polypeptide is necessary for the expression of the heterotropic interactions of the native pigment.


Conformational changes in human serum albumin around the neutral pH from circular dichroic measurements. The molar ellipticity of the warfarin-albumin complex at 310 nm increases with pH from 6 to 9. This pH dependence runs parallel with that of the molar ellipticity of the albumin alone at 292 nm. The change in molar ellipicity with pH occurs in a smaller pH interval after addition of the physiological concentration of calcium ions. These findings give support to the assumption that the binding site for warfarin on the albumin molecule is affected by the neutral-to-base transition in the protein.


[Purification and physico-chemical properties of collagenase synthesized by a bacterium of the type Acinetobacter sp]. The Acinetobacter spec collagenase has been almost completely purified. This enzyme is a true collagenase the activity of which is high on collagen. The enzyme is active on insoluble collagen, gelatin and the synthetic Pz-peptide, but has no proteolytic activity on casein or bovine serum-albumin. The collagenase was obtained on a simple medium with gelatin and yeast extract. The enzyme was purified by (NH4)2SO4 precipitation. DEAE cellulose column chromatography, Sephadex G 200 gel-filtration. The molecular weight of the enzyme was found to be 102 000 daltons, and its isoelectric point was found to be 7,7 +/- 0,2. The optimum pH and temperature for insoluble collagen hydrolysis were 7.6 and 37 degrees C, respectively; so, this collagenase corresponds to true collagenase. Hydrolysis of Pz-peptide is activated by Ca2+ and inhibited by metal ions (Cu2+, Fe3+, Zn2+, Pb2+, Hg2+). EDTA and o-phenanthroline induced a very significant reduction in enzyme activity. Iodoacetate and p-CMB induced a slight reduction in enzyme activity only at high concentrations (10-2M). The collagenase is most stable for temperatures less than or equal to 50 degrees C.


Simultaneous transcutaneous oxygen tension and tissue pH monitoring in sick neonates. Our studies show that simultaneous tcPO2 and tpH monitoring is feasible and they indicate a highly significant degree of correlation between arterial PO2 and tcPO2 on the one hand and arterial pH and tpH on the other. We also conclude that tissue pH as measured with tpH electrode is a close approximation of arterial pH in sick neonates.


Pharmacotherapeutic trials in tardive dyskinesia. Some of the clinical factors contributing to the currently unsatisfactory state of therapy for tardive dyskinesia are reviewed. Problems such as lack of clear syndrome delineation and phenomenological description, the lack of standardization in rating scales and the lack of attention to trial design have all probably contributed to a rather confusing picture. Controlled trials suggest that several pharmacological agents may be of therapeutic value but that clinical prediction of an individual's response is impossible. The strategy of acute drug challenge has emerged as perhaps the most promising approach to the definition of pharmacological subtypes and therefore the choice of optimal treatment.


Tardive dyskinesia and dementia.

Tardive dyskinesia and dementia. Seventeen schizophrenic patients with tardive dyskinesia (TD) and 33 schizophrenics without tardive dyskinesia were examined by psychological tests of intellectual function and EMI scans were performed. The group as a whole were found to be demented and 31 out of 45 had abnormalities on the scan. On a learning test the tardive dyskinesia group did significantly worse and using a measured parameter of the scan (the Ventricular Index) the tardive dyskinesia group had more abnormality. It is suggested that the higher incidence of pathology in the tardive dyskinesia group may be related to chronic neuroleptic toxicity.


Metoclopramide and haloperidol in tardive dyskinesia. The effect of single intravenous doses of metoclopramide (10 mg, 20 mg and 40 mg) and haloperidol (5 mg and 10 mg) have been compared to placebo (saline) in a double blind randomised study in 8 patients with tardive dyskinesia secondary to neuroleptic therapy. Tardive dyskinesia rating scores were improved significantly (P less than 0.01) 6 hours after dosing by metoclopramide 40 mg, and haloperidol 5 mg and 10 mg, when compared to placebo. Single doses of dopamine receptor blocking agents improve tardive dyskinesia. The dose of metochlopramide required to show a beneficial effect was high, and this therefore suggests that it is unlikely to be of therapeutic value as the incidence of adverse reactions would be greatly increased. By monitoring the effects of single doses of dopamine receptor blocking drugs in patients with tardive dyskinesia it is possible to compare the relative potencies of these drugs on dopaminergic systems in vivo in man.


Minimal surgical interference in the prune belly syndrome. Twenty-seven cases of prune belly syndrome which were well in the neonatal period are reported. Eleven were seen in the first year of life and 16 presented later. The first line of treatment was to ensure proper bladder emptying, usually by urethrotomy. Thereafter a policy of minimal surgical interference was followed, operating only for proven obstruction and intractable infection. This policy appears justified as 10 of 11 early presenters have done well. In the late presenters it was accepted that some renal damage had already occurred; nonetheless, 11 of 16 cases have done well. The X-rays were reviewed and hitherto unreported medullary cysts were noted. No radiological signs were found that were of prognostic significance.


[Peritoneal vascular injuries induced by gaseous oxygen]. When introduced as a gas into the abdominal cavity of the rat, oxygen above the initial concentration of 10 % provokes vasodilatation, perivascular oedematous infiltrations and fluid formation. These lesions are entirely reversible. Antihistaminics (promethazine and mepyramine 2 mg/100 g completely inhibit vascular lesions.


The function of fimbriae in Myxococcus xanthus. I. Purification and properties of M. xanthus fimbriae. Myxococcus xanthus fimbriae have been purified and characterized as part of a study of the function of fimbriae in this prokaryote. Myxococcus xanthus produced two types of fimbriae, termed flaccid (F) and rigid (R) on the basis of electron microscopy. F and R fimbriae differed slightly in their response to pH and freeze-thaw regimes but were similar in their resistance to hydrolytic enzymes, amino acid composition, molecular weight, carbohydrate content, and antigenic determinants. Although the precise relationship between F and R fimbriae is unknown, the possibility is considered that F fimbriae might represent a "contracted" form of the R type. Studies designed to determine fimbriae function in M. xanthus are described in an accompanying report.


Biochemical studies on pili isolated from Pseudomonas aeruginosa strain PAO. Pseudomonas aeruginosa strains PAO and PAK bear polar pili which are flexible filaments having a diameter of 6 nm and an average length of 2500 nm. Both types of pili are retractile and promote infection by a number of bacteriophages. The present communication describes the partial biochemical characterization of PAO pili isolated from a multipiliated nonretractile mutant of PAO. The observed properties are compared to those of PAK pili which were characterized previously. PAO pili were found to contain a single polypeptide subunit of 18 700 daltons. This is similar to PAK pili which contain a single polypeptide of 18 100 daltons. The amino acid composition of PAO pilin was also similar to that of PAK pilin. Neither protein contained phosphate or carbohydrate residues and both were found to contain N-methylphenylalanine at the amino terminus. Sequencing of 20 amino acid residues at the amino terminal end of PAO pilin revealed the sequence to be identical with that of PAK pilin, while tryptic peptide analyses of PAO and PAK pilin indicated that the two proteins probably contain a number of homologous regions within the polypeptide. It was concluded that PAO and PAK pili were closely related structures.


[Effect of 2-ketogluconic acid synthesis on the exopolysaccharide production in a Rhizobium meliloti strain]. Two categories of carbon substrates are defined for Rhizobium meliloti: the first favours the synthesis of exopolysaccharides (fructose belongs to this category) while the other is not suitable (glucose belongs to this category). With fructose, resting cells synthesize polysaccharides during more than 100 h and this synthesis is at its best in aerobic conditions at 30 degrees C. With glucose, 2-ketogluconic acid accumulates and rapidly stops the synthesis. The method used to stop this acidification allows with glucose a synthesis which can be compared to the one obtained with fructose.


The function of fimbriae in Myxococcus xanthus. II. The role of fimbriae in cell-cell interactions. Anti-fimbriae antiserum specifically inhibited swarming but no gliding motility per se in Myxococcus xanthus. However, formation of motile aggregates on agar and clumps in liquid media correlated with the presence of fimbriae. Ethylenediaminetetraacetic acid which inhibited swarming also inhibited fimbriae formation. Direct electron-microscopic observations revealed that fimbriae establish contact with apposing cell surfaces. Intact but not depolymerized fimbriae exhibited hemagglutination activity against guinea pig erythrocytes. This activity was inhibited by mannose, N-acetyl-D-galactosamine, and to a lesser degree by fructose, raffinose, melibiose, and alpha-methyl-D-mannoside. It is concluded that fimbriae are organelles which function to establish and maintain intercellular contacts, perhaps by a lectin-like function, during the coordinated movement of cell aggregates' (swarming) in myxobacteria. This hypothesis is supported by the observations of other workers that genes determining movement of cells in groups also control fimbriation in M. xanthus.


Purification and some properties of uridine diphosphate N-acetylglucosamine pyrophosphorylase from Neurospora crassa. Uridine diphosphate N-acetylglucosamine pyrophosphorylase (EC. 2.7.7.23) of Neurospora crassa has been purified approximately 210-fold with dithiothreitol as the stabilizing agent by use of chromatographic techniques. The enzyme preparation appeared to be homogeneous when subjected to electrophoresis. The molecular weight was estimated as approximately 37 000 by gel filtration. The enzyme had an isoelectric point around pH 4.4. Maximum activity of the enzyme was observed at pH 7.5. The enzyme required Mg2+, which may be replaced by other divalent cations such as Mn2+ and Co2+ for lesser degrees of effectiveness. The enzyme was strictly specific for UDP-N-acetylglucosamine as the substrate. The estimated values of Km were 2.2 mM for UDP-N-acetylglucosamine and 5.4 mM for inorganic pyrophosphate. The enzyme activity was highly stimulated by the addition of dithiothreitol or dithioerythritol but was lost by sulfhydryl inhibitory reagents.


Cellulose degradation by a new isolate from sewage sludge, a member of the Bacteroidaceae family. A mesophilic anaerobe, a member of the Bacteroidaceae family (NRC2248), isolated from a cellulose-enrichment culture, digested untreated Whatman cellulose powder and HCl-treated cotton battings while producing hydrogen, carbon dioxide, cellobiose, glucose, and acetic acid as the sole volatile acid. This organism also utilized cellobiose as carbon and energy source but did not utilize glucose. It grew well in synthetic medium containing ammonium salts as nitrogen source and having a pH value of 7.0-7.1 and an Eh value of -160mV or lower. The nutrient requirements of this organism were found to be similar to those of other anaerobes except for Na2S which inhibited cellulose degradation in concentrations above 0.75 mM. Best cellulose degradation occurred under an atmosphere of 80% N2-20% CO2. Use of H2 or 80% H2-20% CO2 as headspace gas inhibited growth. Although accumulation of acetic acid in about 15-16 mM concentrations inhibited the further formation of H2, CO2, and acetic acid in the broth, it did not stop the degradation of cellulose. The results indicate that this organism has the ability to grow in media containing up to 20 g/L of cellulose and to produce industrially important and easily separable end products from cellulose.


Asparagine metabolism and asparaginase activity in a euryhaline Chlamydomonas species. A Chlamydomonas species isolated from a marine environment possesses an L-asparaginase, an enzyme not yet reported in the microalgae. This enzyme enabled the organism to grow as well with asparagine as sole nitrogen source as with inorganic nitrogen sources (NO3-, NH4+). Only the amide nitrogen was used for growth since growth did not occur on aspartate and aspartate accumulated in the media when cells were either grown on asparagine or during short-term incubations with L-[U-14C]asparagine. Cells grown on NO3-, NH4+, or L-asparagine in batch culture possessed equivalent asparaginase activities. However, nitrogen-limited cells possessed four times the activity of cells grown with sufficient nitrogen for normal growth, regardless of the possessed the lowest activity per cell, while lag phase and stationary phase cells possessed greater activity. The enzyme behaved like a periplasmic space enzyme since (1) breaking the cells did not release into solution more activity than was shown by whole cells and (2) whole cells converted L-[U-14C]asparagine to [14C]aspartate with little intracellular accumulation of radioactivity. Cell-free preparations of the enzyme possessed a Km value for asparagine of 1.1 x 10-4 M, with no glutaminase activity.


The effect of changes in pH on phosphate and potassium uptake by Monascus rubiginosus ATCC 16367 in submerged shaken culture. Monascus rubiginosus ATCC 16367 was cultivated aerobically in media containing ammonium and nitrate as nitrogen source. The pH of the medium was adjusted at different times, the pH of the nitrate medium being lowered to the pH of the ammonium medium and the pH of the ammonium medium raised to that of the nitrate. More phosphate was taken up on the nitrate medium, but potassium uptake did not start until 24h. On the ammonium medium, both were taken up in parallel from the beginning, but the amount of phosphate taken up never reached the same level as on nitrate medium. When the pH was adjusted, the uptakes changed, especially on the ammonium medium where a great increase in phosphate uptake was observed. More conidia were formed on the nitrate medium and more pigment on the ammonium medium. When the pH of either media was adjusted, the development of conidia and pigment production changed to that of the other control medium where the pH evolved normally in the direction of the change, regardless of the source of nitrogen. The reasons for the development of conidia on nitrate medium or where the pH is high, and the production of pigment on ammonium medium or at low pH is discussed.


Virological surveillance for dengue haemorrhagic fever in Indonesia using the mosquito inoculation technique. A dengue haemorrhagic fever surveillance system in Indonesia, based on virological and clinical observations, is described. The system uses the mosquito inoculation technique for virus isolation and is simple, economical, and well suited for endemic areas where support and facilities are limited. The data suggest that with good cooperation between the hospital and the virology laboratory, new serotypes and possibly even new strains of virus can be identified before the onset of epidemic activity. This type of virological surveillance may make it possible to prevent major epidemics in the future.


Daily fluctuation of cholestatic enzymes in bile and its relationship to the total bile salts concentration. The activities of alkaline phosphatase (AP), gamma-glutamyl-transpeptidase and leucine aminopeptidase as well as the total bile salt concentration (Tbs) were measured in bile specimens collected spontaneously at 7 and 11 a.m. and 4 and 9 p.m. from the 3rd-9th postoperative day of 16 patients with T-tube insertion. The general trend for all cholestatic enzymes to increase or decrease was in some way related to the total bile salt concentration. The best correlation was found between AP and Tbs (r = 0.48). It is suggested that the enterohepatic circulation of bile salts may lead to the delivery of membrane-bound cholestatic enzymes into the bile canaliculi without damage to the hepatocyte.


Interaction of gamma-glutamyltransferase from human tissues with insolubilized lectins. We have characterized the binding of gamma-glutamyltransferase to three insolubilized lectins. Optimal binding was achieved in 2 hours at 25 degrees C for concanavalin A and at 4 derees C for ricinus communis agglutinin 120 and wheat germ agglutinin,and was also a function of the ratio of lectin protein to gamma-glutamyltransferase protein. The interaction of gamma-glutamyltransferase with these three lectins is specific, and release of bound enzyme by carbohydrates follows the same general order of specificity previously observed for the competition between mono or polysaccharides for the lectin carbohydrate binding sites. The binding of trypsin-solubilized liver gamma-glutamyltransferase to the three insolubilized lectins was virtually identical to that of detergent solubilized enzyme. We propose, therefore, that the release by proteolytic enzymes, of gamma-glutamyltransferase from plasma membrane matrix does not significantly alter its carbohydrate structure. We obtained great differences in binding to the three lectins between the liver, kidney, pancreatic and duodenal isoenzymes of gamma-glutamyltransferase. From this data we conclude that carbohydrate content and topography are important distinguishing features of gamma-glutamyltransferase isoenzymes.


Predictive values of various liver function tests with respect to the diagnosis of liver disease. A prospective study of 181 patients suspected of having liver disease was carried out to determine the relative efficiencies of serum bilirubin (total and direct), alkaline phosphatase (AP), gamma glutamyl transferase (GGT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) with respect to diagnosis. Liver biopsies, liver scans, abdominal ultrasound, and clinical parameters were also tabulated and used independently to evaluate the patient's hepatic status and to determine the final diagnoses in each case. From the results of these tests for the 60 patients who were diagnosed as having liver disease, and the 87 patients who were felt to be free of liver disease, predictive values of the above tests were established. Data from this study suggests that while direct bilirubin is the most specific test, GGT is the most sensitive and has the fewest false negatives in the diagnosis of liver disease.


Prolactin release by intravenous cimetidine in man: evidence for a suprapituitary locus on action. In an attempt to identify the sites at which cimetidine stimulates prolactin release, the drug was administered intravenously (6 mg/kg body weight) to healthy subjects under basal conditions, during dopamine infusion (1 microgram/Kg-min for 120 min) and after pretreatment with L-dopa plus carbidopa (250 plus 25 mg every 6 for 1 day). The serum prolactin response to cimetidine was abolished by dopamine infusion and almost completely suppressed by L-dopa plus carbidopa administration. These findings suggest that the drug acts on the central nervous system to stimulate prolactin release. Although the mechanism of this action is unclear, it does not seem to depend on an antidopaminergic effect and may be related to blackade of brain H2 histamine receptors.


Distribution of beta-adrenergic receptors on human lymphocyte subpopulations. A technique is described allowing the quantification and the characterization of specific beta-adrenergic receptors in intact living human lymphocytes. 125I-Iodohydroxybenzylpindolol, a potent beta-adrenergic antagonist was used to label specific binding sites on unfractionated lymphoid cells and on purified subpopulations of T (F1 and F2) and B cells. F1 and F2 were obtained by filtration through nylon wool column as previously described (Delespesse et al., 1976), they differ in their response to mitogens, and in their interactions with adherent cells and B cells. 125I-HYP binding to unfractionated lymphocytes was a saturable, stereospecific and rapid process with a dissociation constant of 2.5 10(-10) M and a binding capacity of 400--600 sites/cell. Bindings on unfractionated lymphocytes, purified B cells and T cells of the F2 fraction were similar. No detectable binding was noted on T cells from the F1 fraction. Enriched T cells obtained by a rosetting technique displayed 200 receptors/cell.


Allogeneic bone marrow transplantation for severe aplastic anaemia--the London experience. Using the Seattle protocol with minor modifications, 23 patients with severe aplastic anaemia received allogeneic bone marrow transplants from HLA/mixed leucocyte culture matched sibs in three London centres between 1973 and 1977. Ten patients (43.5%) are alive 6 months to 5 years after transplantation, and are well with full haemopoietic reconstitution, two with autologous bone marrow recovery following the graft procedure. A failure of the marrow graft to take, or take followed by rejection occurred in 12 patients (52%). Failure of marrow recovery was associated with a high early mortality from bacterial or fungal infection. The only survivors amongst those who rejected the first graft were four patients in whom a subsequent graft from the same donor was successful, and two in whom autologous recovery occurred. Graft versus host disease (GVHD) occurred in seven patients, and was fatal in one case. The most frequent complication after successful engraftment was varicella-zoster infection which occurred in five patients and was fatal in one patient. The overall results compare favourably with those from other transplant centres, but the high rate of graft rejection and low incidence of GVHD differ from other series. The results should encourage further referral of patients with severe AA for bone marrow transplantation.


Irreducible dislocation of the metatarsophalangeal joints of the foot. An unusual dislocation of the metatarsophalangeal joints of the third, fourth, and fifth toes with fractures of the neck of the second metatarsal bone occurred in a 23-year-old man. Hyperextension of the metatarsophalangeal joints is the mechanism of injury and is substantiated by the anatomic investigation. The metatarsal head is trapped between the fibrocartilaginous plate on the plantar surface of the foot, the dorsal capsule and deep transverse metatarsal ligament on the dorsal surface, the flexor tendons on the lateral aspect and the lumbrical tendons on the medial aspect. Division of the fibrocartilaginous plate, the deep transverse ligament and the dorsal capsule facilitates reduction of dislocation.


Behavioral toxicity. Neuroleptic-induced paradoxical behavioral toxicity in young borderline schizoprenics. 9 young borderline schizophrenics who developed an acute paradoxical behavioral toxicity as a reaction to the initiation of antipsychotic medication are described. The dynamics, the psychological issues as well as some possible psychopharmacological interactions are discussed. The risk of behavioral toxicity should not be overlooked or underestimated and the occurrence of this reaction must be carefully differentiated from other drug-induced reactions.


The effects of rubidium on mammary tumour growth in C57 blk/6J mice. A high pH therapy for cancer arrived at theoretically was tested in mice by feeding them rubidium carbonate. Tumours were transplanted in the abdomen of mice and allowed to grow for 8 days. The mice were then divided into two groups. The control group was continued on conventional mouse chow. The test group, in addition to the mouse chow, was force-fed 1.11 mg of rubidium carbonate dissolved in distilled water. At the end of 13 more days the tumours in the controls had grown to a large size so all the mice were sacrificed. The tumours were then removed and weighed. The tumours in the test animals weighed essentially one eleventh of those in the controls. In addition the test animals were showing no adverse effects from the cancers. The probability that this marked difference in tumour size could have come about by chance is exceedingly small.


Brain regional levels of neurotransmitter amines as neurochemical correlates of sex-specific ontogenesis in the rat. Brain regional levels of three neurotransmitter amines - serotonin (5-HT), norepinephrine (NE), dopamine (DA) - were measured in young rats prior to weaning to determine the extent to which modifications in levels of amines might reflect alterations in the sex steroid hormonal environment during the first postnatal week in the life of the rat. Sex-related levels of DA, NE, and 5-HT were found in some brain regions of the 12-day-old rat. Male midbrain DA exceeded the corresponding female value while female hypothalamic NE levels were greater than those of the male. Levels of 5-HT in the corpus striatum and the midbrain of males were greater than those of the female. Castration of the male on day 1 or testosterone propionate (TP) administration to the newborn female resulted in modifications of levels of midbrain 5-HT which reflected feminization of the castrated males and masculinization of the TP-treated females. Castration on day 1, or diethylstilbestrol given on days 2, 4, and 6, resulted in apparent feminization of NE levels in the hypothalamus of 12-day-old male rats. Thus, it appears that regional levels of hypothalamic NE and midbrain 5-HT in the 12-day-old rat may reflect the course of brain organizational activity which becomes recognizable in the adult as sex-specific behavior.


Arterial or venous umbilical pH as a measure of neonatal morbidity? Although umbilical arterial pH is usually used as a parameter of fetal asphyxia, blood samples are more easily obtainable from the umbilical vein. The relationships of both venous and arterial pH with Apgar scores and neonatal neurological assessment were compared. There was a good correlation between venous and arterial pH and the relationship between venous pH and Apgar scores was similar to that between arterial pH and Apgar scores. Venous pH did not appear to be inferior to arterial pH and Apgar scores in predicting neonatal neurological morbidity.


The pathochemistry of kernicterus.

The pathochemistry of kernicterus. The stoichiometry of bilirubin--albumin interaction has been analyzed and quantitated in several recent studies, confirming that albumin binding of bilirubin obeys the law of mass action [4, 5, 14, 16, 26, 36, 43, 46, 61, 65, 73, 92, 111]. These studies provide a basis for interpreting bilirubin transport, cell uptake and toxicity from physicochemical and pharmacologic perspectives [35, 42, 58, 59]. In this report, we propose a model of the pathogenesis of kernicterus which views serum albumin and tissue as competing with each other for binding the miscible bilirubin pool. Evidence is presented to show that bilirubin normally binds reversibly to cellular membranes and certain soluble enzymes just as it does to albumin; the unbound bilirubin concentration is the driving force for both albumin and tissue binding. We propose that albumin binding is determined by the concentration of free bilirubin anion (which is essentially unaffected by physiologic pH changes), and that tissue binding is mainly determined by the concentration of free bilirubin acid (which is greatly influenced by pH). When bilirubin--tissue complexes are formed, essential cell functions may be inhibited, producing cellular acidosis, irreversible intracellular aggregation of bilirubin, and cell death. In developing this argument, we will sequentially discuss relevant features of bilirubin chemistry, the binding of bilirubin to albumin, the formation of bilirubin--tissue complexes, bilirubin toxicity, alternative viewpoints of bilirubin transport, and, finally, the implications of this model to the clinical management of jaundiced infants. It should be emphasized that this paper is an attempt to analyze bilirubin transport and toxicity using basic chemical principles; it is an extension of previously published proposals [17, 77], and will undoubtedly require further modification as additional experimental data becomes available.


The pharmacokinetics and biotransformation of the new benzodiazepine lormetazepam in humans. I. Absorption, distribution, elimination and metabolism of lormetazepam-5-14C. The pharmacokinetics and metabolism of the new benzodiazepine lormetazepam were investigated in five male volunteers using the 14C-labelled drug (position 5). Lormetazepam was administered intravenously and orally, at a dose of 0.2 and 2 mg respectively, to each of the test subjects. Measurements of total radioactivity showed that the drug was absorbed completely and eliminated almost exclusively by the renal route. Maximum plasma level of active ingredient and total radioactivity were observed about 2 hours and 5 hours following oral administration. As early as 30 min following oral administration, concentration of active ingredient amounted to 80% of the maximum values. After both treatments the terminal half-life of total radioactivity and lormetazepam glucuronide in plasma corresponded to the half-life of elimination in urine of about 13 hours. After enzymatic hydrolysis with beta-glucuronidase/arylsulphatase, an average of 90% of total radioactivity from various urine and plasma samples was extractable with ether. Extracts from plasma contained only unchanged drug, indicating free and conjugated lormetazepam as ingredients of total radioactivity. Extracts from urine could be separated into lormetazepam and its N-demethylation derivative lorazepam. The relative amount of excreted lorazepam conjugate was demonstrated to be time-dependent, probably due to enterohepatic circulation. Since less than 6% of the total dose was demethylated by both routes of administration, it can be assumed that lormetazepam is the active product.


The N protein of bacteriophage lambda, defined by its DNA sequence, is highly basic. Nucleotide sequence has been determined for the restriction fragments and cloned DNA from the pL-N-tL1 region of bacteriophage lambda. A unique reading frame for the N gene is defined by the absence of natural nonsense codons and by the presence of seven nonsense codons generated by mutations in N. This reading frame is initiated at two alternative ATG codons, the second of which is probably the in vivo translation start. Reading is stopped at a single TAG codon. The protein coded is therefore 133 or, more probably, 107 amino acids long, rich in lysine, arginine and proline.


Fetal heart rate in the second stage of labour. Method of interpretation and parameters of the new-born. The authors present a protocol for the interpretation of the fetal heart rate during the second stage of labour (limited to the last 30 min before delivery). The FHR recordings were grouped according to five classes of increasing pathological significance. Good correlations were obtained between these classifications and the Apgar score as well as the umbilical artery pH.


Proteinase activity and agglutination of leukemia cells. The proteinase activity was assayed in the leukemia cells L 1210 and in the ascites fluid with [3H] acetylated haemoglobin as a substrate. The proteinase activity at pH 4.1 increased in cells and in the ascites fluid with age of the tumor. The proteinase activity at pH 7.8 was low, but the enzyme activity in the cell homogenate increased between 5th and 7th day of the tumor growth and it was also present in the ascites fluid. It was observed that the leukemia cells aggregate in vivo and in vitro at pH values of the ascites fluid above pH 7.0. It was suggested, that the aggregation of leukemia cells is due to the tumor cell proteinase activity released to the ascites fluid.


Influence of sex hormones on gamma-glutamyltranspeptidase activity in rats serum with Morris hepatoma 5123 D. The Stilbestrol treatment of the female rats with the Morris hepatoma 5123 D resulted in decrease of the gamma-glutamyl-transpeptidase (GGTP) activity in serum and in hepatoma, but only during the treatment. Given to the male rats under the same conditions, Stilbestrol had no influence on the GGTP activity. Castration of males was the cause of the GGTP activity decay in hepatoma and in serum, but it was not the case with females. Sustanon diminished the GGTP activity in serum of the female rats with hepatoma.


[Pharmacological properties of procaterol, a newly synthetized, specific beta 2-adrenoceptor stimulant. Part I. Effects on the CNS (author's transl)]. Effects of procaterol (PRO) on the CNS were investigated in comparison with those of salbutamol (SAL) and isoproterenol (ISO). PRO, 15 to 50 mg/kg given subcutaneously suppressed spontaneous movement in mice, rats and rabbits and with a large dose, 1000 mg/kg, the animals became quiet and immobile. In dogs, PRO produced similar symptoms and in addition, there was nausea and vomiting. The animals recovered within 3--8 hours. ID50's in depressing spontaneous movement were 20.2 and 245 mg/kg for PRO, 51.1 and 133 mg/kg for SAL and 2.37 and 143 mg/kg for ISO, respectively, both by the subcutaneous and oral routes of administration. Methamphetamine induced increase in motility and fighting behavior was also suppressed by PRO when similar doses were given. PRO had no effect on coordinating movement, halothane anesthesia, drug and electric stimulation induced convulsions and body temperature, and there was no muscle relaxant action. However, PRO in large doses prolonged sleeping time with hexobarbital. The analgesic effect of PRO was not observed with Haffner's and Landall Selitto's methods but acetic acid induced writhing was suppressed by PRO. PRO had little effect on spontaneous EEGs either cortical or from deep structures, and EEG arousal responses. The effects of PRO on the CNS were slight and nonspecific, and similar to those of SAL and ISO.


[Pharmacological properties of procaterol, a newly synthesized, specific beta 2-adrenoceptor stimulant. Part II. Effects on the peripheral organs (author's transl)]. Pharmacological properties of procaterol (PRO) in the peripheral organs were examined in comparison with those of sulbutamol (SAL) and isoproterenol (ISO). PRO slightly enhanced twitch tension of the tibialis anterior muscle but affected little the mono- and poly-synaptic spinal reflexes and ganglionic transmission. PRO depressed spontaneous contractions of the isolated ileum, non-pregnant and pregnant uterus and also the gastrointestinal and uterine movements in vivo. PRO prolonged the time of peroral charcoal transport in the intestine. Potencies of PRO in producing these effects were between those of ISO and SAL except those on the uterus in which PRO was more potent than ISO and SAL. Pro depressed gastric and bile secretion but had no effect on pancreatic secretion. ISO, PRO and SAL reduced resistance of the common carotid, femoral and renal arteries and the relative potencies of PRO and SAL to ISO were significantly less in the renal artery than in the other arteries. In accelerating heart rate in conscious rats and dogs, PRO (p.o. or s.c.) was almost equipotent to SAL. Urine flow, GFR, RPF, free water and osmolar clearance and also excretion of electrolytes were reduced by PRO with the concomitant fall of systemic blood pressure. PRO has no effect on blood coagulation and hemolysis but inhibited carrageenin edema and an increase in permeability of blood vessels induced by acetic acid. PRO had no alpha-adrenolytic, cholinolytic and anti-histaminic effects.


[The antiallergic effect of Balkis. A rhino-manometric study]. The antiallergic and decongestive effects of the common cold preparation Balkis was tested by rhinomanometry in human volunteers usually suffering from allergic rhinitis after oral treatment. The nasal irritation in these patients was induced by administration of the specific allergen. This kind of test is especially appropriate for the control of drugs in cases of ordinary colds, as the test conditions are reproducible in symptomatically similar cases. The rhinomanometrical control of the respiratory resistance was carried out before and after an allergenic exposal as well as before and after administration of Balkis and allergenic exposal in 30 patients (17 females, 13 males; 9-49 years old). It is shown that Balkis had a good to satisfactory antiallergic effect in 67% of all cases. In 33% of the cases the antiallergic effect was not evident. This result confirms objectively the good antiallergic effect of this common cold preparation by oral administration. Balkis was well tolerated in all of the 30 patients. The symptoms of slight fatigue which appeared in 1/3 of all cases were only transitory.


[Azathioprine in rheumatology].

[Azathioprine in rheumatology]. The indication for azathioprine (Imurek) has to be strictly adjusted, but amongst the immunosuppresive drugs it is without doubt the best tolerated substance and--provided that regular controls are carried out--is therefore also suitable for long-term treatment in practice.


Urinary adenosine 3',5'-monophosphate (cyclic AMP) in normal and cryptorchid boys. The 24-hour urinary excretion of cyclic AMP was determined in 102 normal boys aged 1.9-16.9 years and in 136 cryptorchids aged 2.5-16.9 years. A marked increase of the normal cyclic AMP excretion was found in pubertal years. There was a positive correlation between urinary excretion of cyclic AMP and the excretion of testosterone, androstenedione, LH and FSH. A positive correlation was also found between cyclic AMP excretion and height and weight, respectively. Mean cyclic AMP excretion of bilateral and unilateral cryptorchids was normal in all bone age groups except in unilateral cases with bone age 8-9.9 years and bone aged greater than or equal to 14 years. In these two groups, mean cyclic AMP excretion was moderately increased. After HCG stimulation of 25 cryptorchids, urinary cyclic AMP excretion varied between increased, unchanged and decreased values. The cyclic AMP excretion changes observed in some of our patients were difficult to interpret and were possibly of unspecific nature. Further information about the testiclar cyclic AMP secretion and the relationship between this nucleotide and sexual hormones may be obtained from studies in testicular biopsy tissue.


Computer-controlled bedside monitoring and infusion system. A mult-channel system for continuous bedside monitoring of glucose, pH pCO2 and automatic computer-controlled infusion of insulin, glucose, potassium, and bicarbonate solutions is described. Regulatory algorithms are presented for correction of metabolic acidosis and hyperglycemia with and without acidosis.


Hydrogen-tritium exchange titration of the histidine residues in bovine heart cytochrome c and analysis of their microenvironment. Microenvironments of the three histidine residues located at the positions 18, 26, and 33 from the amino terminus in bovine heart cytochrome c were analysed in solution by the hydrogen-tritium exchange titration method, which has been developed in this laboratory. Histidine-18, which is liganded to the heme iron, and histidine-26 did not incorporate tritium in native state, indicating that the two are located in solvent inaccessible hydrophobic regions. Histidine-33 was labeled with tritium to an appreciable extent and seemed to be partially buried in the molecule. The pKa value estimated for histidine-33 was 6.1 at 37 degrees by the tritium exchange titration, suggesting that the residue interacts very weakly with a neighboring cationic group. These results seem to be compatible with the tertiary structure of the protein deduced from the X-ray crystallographic analysis.


Formation of aminosuccinyl peptides during acidolytic deprotection followed by their transformation to piperazine-2,5-dione derivatives in neutral media. Prolonged acidic treatment of Boc-Leu-Asp(OBut)-Phe-NH2 with 4N HCl in acetic acid resulted in H-Leu-Asc-Phe-NH2 . HCl(Asc, aminosuccinyl), which transformed partially to cyclo[Leu-Asp(Phe-NH2)] during its purification by column chromatography on silica gel with a mixture of ethyl acetate/pyridine/acetic acid/water = 60:20:6:11, i.e. in neutral medium. Examination of the imide formation was extended to different reaction conditions (no imide derivative was detected in trifluoroacetic acid), to several protected derivatives of L-aspartyl-L-phenylalaninamide and to tripeptides containing an aspartyl residue in the middle position. It was clearly demonstrated that in strongly acidic media the imide derivatives are directly formed from the aspartyl peptides containing a free beta-carboxyl group. The influence of the C-terminal residue was greater than the N-terminal on both the rate of formation of the imide and its further transformation to piperazine-2,5-dione derivative. In aqueous ethanol the X-Asc-Y-NH2 (X, Pro, Leu; Y, Gly, Ala, Val, Phg, Phe) containing N-terminal proline are more readily transformed to piperazine-2,5-dione derivatives, but compared to simple proline dipeptides the rate of this transformation is relatively slow because of the crowdedness of the tricyclic transitional state.


[Discontinuation of antipsychotic drugs in chronic schizophrenic patients. II. Psychological and psychopathological test differences between patients with and without relapse]. After the discontinuation of the neuroleptic treatment (open design), 10 out of 30 hospitalized chronic schizophrenic patients showed before the end of 4 weeks a significant increase of their symptomatology. This deterioration made the application of neuroleptics necessary. Before the discontinuation of the neuroleptic treatment the group of patients with later relapse and the group without relapse showed only slight differences concerning psychopathological symptoms but significant differences concerning test performance. After 2 weeks without medication a pronounced impairment of performance was found in patients with later relapse. At this time the two groups showed no more significant differences in the psychological tests. At the same time significant differences concerning the psychopathological symptoms had developed.


[Necrotizing vasculitis: its manifestation as urticaria]. A case of urticarial vasculitis, a distinct form of necrotizing vasculitis is described. This entity is characterized by a chronic, persistent urticaria with petechiae and the histological features of necrotizing vasculitis. Serum complement is depressed and multisystem involvement can often be found. There is evidence for immune complexes to play a role in the pathogenesis of this disease.


Mucous glycoproteins of teleostean fish: a comparative histochemical study. To examine the hypothesis that the histochemical characteristics of teleostean mucus reflect functional characteristics, mucous cells were studied in four related and behaviourally similar species of fish (Family Belontidae). Histochemical characteristics were determined with Alcian Blue at both pH 2.6 and pH 1.0 followed by the periodic acid-Schiff technique. It was found that the four species differed in glycoprotein type as well as in number of mucus-containing cells. The differences are discussed in regard to functional characteristics and environmental influence.


Enzyme histochemical characteristics of human and kitten odontoclasts and kitten osteoclasts: a comparative study using whole cells. Methods for the histochemical demonstration of enzymes in whole cell preparations of odontoclasts and osteoclasts are described. Enzyme histochemical characteristics of human and kitten odontoclasts from resorbing primary teeth and of osteoclasts from kitten femur metaphyses were determined and compared. The enzyme profiles, times for the appearance of detectable reaction product, intensity of the reactions and localization of the reaction products were similar in all three types of giant cell. These findings suggest that odontoclasts have enzyme properties and metabolic functions similar to those of osteoclasts. Species differences appear to be minor, although the NADP-dependent enzymes are less active in human than in kitten odontoclasts. Both odontoclasts and osteoclasts are rich in enzymes concerned with energy production and possess considerable activity of enzymes usually associated with catabolic functions. Metabolic pathways are well developed in respect of the utilization of succinic, malic, glutamic, lactic and isocitric acids, beta-hydroxybutyric acid and glucose-6-phosphate, and they also possess phosphatases, non-specific esterases and leucine naphthylamidase. The distribution of enzyme reaction products for the individual enzymes demonstrated is consistent with the presence in these cells of large numbers of mitochondria and lysosome-like organelles. Considerable phosphatase activity is demonstrable in both odontoclasts and osteoclasts at both neutral and acid pH.


Central chemical regulation of respiration in term newborn. The role of the medullary H+-sensitive chemoreceptors on the drive of breathing was studied in 10 unanesthetized newborn animals (8 lambs and 2 kids). The experiment consisted of sequential measurements of ventilation (VE) during a progressive change in the arterial pH (pHa) and in the pH of the cisternal cerebrospinal fluid (pHCSF), induced by intravenous infusion of hydrochloric acid (HCl) followed after an 8-h steady state of acidosis by rapid bicarbonate [HCO3-] infusion. It is shown that a rapid change in [HCO3-]CSF occurs during the infusion of HCl or NaHCO3. As a consequence both CSF and arterial pH change in the the same direction and large changes in pHCSF (from 7.331 to 7.227) were observed. Such CSF acidosis did not contribute to further increase VE beyond the level by hyperventilation induced by the initial fall of pHa. The ventilatory response to the decrease in pHa was found to fall off with moderate to severe acidosis (pHa less than 7.20). In conclusion, this study demonstrates an instability of the pHCSF during neonatal metabolic acidosis and it suggests an immaturity of both the H+-sensitive medullary and peripheral chemoreceptors in the 8-day-old newborns.


Reversed-phase high-performance liquid chromatography of catecholamines and their congeners with simple acids as ion-pairing reagents. We have investigated the value of various common acids as ion-pairing reagents for high-efficiency separations of catecholamines and their metabolites in reversed-phase high-performance liquid chromatography. The retention of norepinephrine, alpha-methylnorepinephrine, dopamine, alpha-methyldopamine, L-dopa, alpha-methyldopa, dihydroxybenzylamine, epinephrine, carbidopa and DOPAC was measured in mobile phases composed of nitric, sulfuric, acetic and trichloroacetic acids at pH 2-5 and anion concentrations ranging from 5-500 mM. The solute capacity ratios were dependent on the hydrophobicity and concentration of the ion-pairing reagent and the pH of the mobile phase. Good retention, peak symmetry and high efficiency (3000 theoretical plates for 300 mm) was found for mobile phases composed of the strong inorganic acids and trichloroacetic acid. Chromatography was compared to that seen using the detergent sodium octylsulfate. Trichloroacetic acid gave retention and efficiency similar to sodium octylsulfate. These experiments show that simple acids can replace alkylsulfates as ion-pairing reagents for the separation of the catecholamines and their metabolites.


Morphology of the antibody response to pneumococcal polysaccharide in mice. In the course of an antibody immune response to pneumococcal polysaccharide - type III (S III) in mice a slight increase was observed in the proportion of plasma cells among the antibody-producing cells, reaching its peak at the time of decline of this reaction. On the basis of ultrastructural resemblance of these plasma cells to primitive reticular cells and in view of other specificities of the immunological response to S III antigen, the author presumes direct reticular origin of anti-S III antibody-producing plasma cells.


Dopaminergic control of oxytocin release in lactating rats. During suckling, anaesthetized lactating rats release regular (about every 7 min) but brief pulses of oxytocin (0.5--1.0 mu.) which produce single transient increases in intramammary pressure. Drugs which selectively impair synaptic transmission were used to determine the role of dopamine and noradrenaline in regulating this natural reflex. Diethyldithiocarbamate (100--200 mg/kg, i.v.) and alpha-methylparatyrosine (100--400 mg/kg, i.v.) which inhibit the synthesis of catecholamines both blocked the suckling-induced release of oxytocin. The milk-ejection reflex was also inhibited in a dose-dependent manner by the intravenous administration of the dopamine antagonists, fluphenazine (0.7 mg/kg), pimozide (1.4 mg/kg), cis-dupenthixol (4.5 mg/kg) and metoclopramide (6.0 mg/kg), and caused a significant inhibition P less than 0.01) of the reflex in 50% of the rats tested. The alpha-adrenoceptor antagonist phenoxybenzamine (1.4 mg/kg) was similarly effective. Dopamine (40 micrograms), bromocriptine (10 micrograms), apomorphine (100 micrograms), noradrenaline (10 micrograms) and phenylephrine (2 micrograms) injected into the cerebral ventricles evoked a sustained release of oxytocin which produced multiple increases in intramammary pressure; isoprenaline (4 micrograms) was ineffective. The release of oxytocin evoked by dopamine and noradrenaline was prevented by cis-flupenthixol and phenoxygenzamine respectively. None of the drugs used affected the mammary sensitivity to exogenous oxytocin nor were their actions modified by pretreatment with propranolol (1 mg/kg). The results suggest that the neural pathway for the reflex release of oxytocin during suckling in the rat contains both dopaminergic and noradrenergic synapses, the latter acting through alpha-adrenoceptors and being distal in the pathway to the dopaminergic component.


Mechanism of inhibition of active potassium transport in isolated midgut of Manduca sexta by Bacillus thuringiensis endotoxin. After incubation at pH 10 or higher, Bacillus thuringiensis spores and endotoxin, at concentrations above 0.1 IU/ml, affected transport parameters in the isolated midgut of Manduca sexta larvae. (Toxic activity was lost during roughly 1 week at pH 11.) About 60% of the short-circuit current was inhibited, and the remainder was reversibly inhibited by anoxia. Electrical resistance was reduced by about 55% and oxygen uptake stimulated by about 30%. Influx of potassium from blood-side to lumen-side ('active' flux) was unaffected but flux in the reverse direction was nearly tripled. These results suggest that hydrolysis of the toxin yields an inhibitor of potassium transport, presumably a polypeptide. It is argued that inhibition is not primarily by uncoupling of oxidative phosphorylation, but instead by interference with an active depression of the efflux of potassium from lumen-side to blood-side.


Pharmacological properties of axonal sodium channels in the cockroach Periplaneta americana L. II. Slowing of sodium current turn-off by Condylactis toxin. Condylactis toxin induces a prolonged falling phase in action potentials recorded from giant axons of the cockroach Periplaneta americana L. A distinct plateau potential can be recorded in the presence of Condylactis toxin at concentrations of 0.5 mg/ml and above. In cockroach axons, voltage-clamp experiments show that Condylactis toxin acts primarily by slowing the sodium current turn-off without affecting either the time-course of the sodium conductance increase or the peak amplitude of the transient sodium current.


Complexes of technetium with pyrophosphate, etidronate, and medronate. The reduction of [99Tc]pertechnetate was studied as a function of pH in complexing media of pyrophosphate, methylene diphosphonate (MDP), and ethane-1, hydroxy-1, and 1-diphosphonate (HEDP). Tast (sampled d-c) and normal-pulse polarography were used to study the reduction of pertechnetate, and normal-pulse polarography (sweeping in the anodic direction) to study the reoxidation of the products. Below pH 6 TcO4-was reduced to Tc(III), which could be reoxidized to Tc(IV). Above pH 10, TcO4-was reduced in two steps to Tc(V) and Tc(IV), each of which could be reoxidized to TcO4-. Between pH 6 and 10 the results differed according to the ligand present. In pyrophosphate and MDP, TcO4- was reduced in two steps to Tc(IV) and Tc(III); Tc(III) could be reoxidized in two steps to Tc(IV) and TcO4-. In HEDP, on the other hand, TcO4- was reduced in two steps to Tc(V) and Tc(III), and could be reoxidized to Tc(IV) and TcO4-. Additional waves were observed; they apparently led to unstable products.


Brain protease activity after experimental head injury. In an experimental series on twelve cats, activity changes of brain cell proteolytic activity were measured two hours after a blunt head injury without hematoma or contusions. Protease activity was estimated in two different brain tissue homogenate supernatants containing total soluble and only cytoplasmic activity without proteases in cell organelles, respectively. Total activity was doubled two hours after injury in the acid and the neural pH-range, in comparison to control values. Free soluble activity was doubled in the acid and increased to the threefold value in the neutral range. From these data, it seems that two different changes appear in lysosomes, the enzyme-reservoir of the cell: (1) Enzyme-synthesis is increased after trauma, measured as augmentation of total soluble protease activity in our experiments; (2) Breakage or increased permeability of lysosomes lead to augmentation of especially neutral proteases in the cytoplasm followed by the well known autolytic areas of generalized traumatic brain edema.


Intracellular pH and the distribution of weak acids and bases in isolated rat superior cervical ganglia. 1. The steady-state intracellular/extracellular concentration ratios (Ci/Co) of a number of radiolabelled weak bases in isolated rat superior cervical ganglia were measured. 2. Observed values for Ci/Co (mean +/- S.E. of mean) were [3H]nicotine, 6.17 +/- 0.12; [14C]morphine, 6.08 +/- 0.14 [3H]atropine, 7.10 +/- 0.16; [14C]trimethylamine, 6.73 +/- 0.13; [14C]procaine, 10.13 +/- 0.26. If Ci/Co were determined by the transmembrane pH gradient, the intracellular pH (pHi) appropriate to these concentration gradients lay between 6.4 and 6.6 at an extracellular pH (pHo) of 7.4. 3. the steady-state value of Ci/Co for the weak acid 5,5-dimethyl-2,4-oxazolidinedione (DMO) was 0.87 +/- 0.007. The appropriate pHi was 7.31 +/- 0.003. 4. The difference between the values of pHi calculated from the distribution of the weak bases and of DMO could not be attributed to (i) experimental error, (ii) partial permeation of protonated base, (iii) intracellular binding or carrier-mediated transport of base, (iv) lipid uptake of base or (v) different pK'a inside and outside cells. 5. The difference between the measurements of pHi made with DMO and nicotine (pHDMO-pHnic) was reduced or abolished by uncoupling agents, which act as transmembrane proton carriers. This effect was not reproduced by respiratory inhibitors or by exposure to lactate. 6. pHDMO-pHnic was small (less than 0.1 units) in human erythrocytes, which contain no intracellular organelles, and was exaggerated (1.0 unit) in slices of lipid-depleted brown adipose tissue which contained an abundance of mitochondria. 7. It is concluded that the different values of pHi determined using weak acids and bases arise from the presence of membrane-bound intracellular compartments of differing pH, and that where the use of pH-sensitive micro-electrodes is impracticable, it is desirable to measure pHi with both a weak acid and a weak base unless these can be shown equal over a wide range of pHi values.


Light induced changes of internal pH in a barnacle photoreceptor and the effect of internal pH on the receptor potential. 1. Intracellular pH (pH1) was measured in Balanus photoreceptors using pH-sensitive glass micro-electrodes. The average pH1 of twelve photoreceptors which had been dark adapted for at least 30 min was 7.3 +/- 0.07 (S.D.). 2. Illumination reduced the recorded pH1 by as much as 0.2 pH unit. The change in pH1 was graded with light intensity. 3. When the cells were exposed to CO2 in the dark, pH1 declined monophasically. Saline equilibrated with 2% CO2; 98% O2 produced a steady reduction in pH1 of about 0.25 unit in 2--3 min. The buffering capacity of the receptor cell cytoplasm calculated from such experiments is approximately 15 slykes. 4. In the presence of HCO3-1, CO2 saline produced smaller, biphasic changes in pH1. 5. The membrane depolarization produced by a bright flash (depolarizing receptor potential) was reversibly reduced in the presence of external CO2 or by injection of H+. Iontophoretic injection of HCO2- increased the amplitude of the receptor potential. 6. In individual cells there was a close correlation between the amplitude of the receptor potential and pH1. 7. Saline equilibrated with CO2 reduced the light induced current (recorded under voltage-clamp) by 40--50% without affecting its reversal potential. 8. Exposure of the receptor to 95% CO2 saline for several minutes (pH0 5.5) not only abolished the receptor potential but also reversibly decreased the K conductance of the membrane in the dark. These effects were not reproduced by pH0 5.5 buffered saline or by a 5 min exposure to saline equilibrated with N2. 9. It is suggested that changes in pH1 induced by light modulate the sensitivity of the receptor under physiological conditions.


Growth and differentiation of Trypanosoma cruzi cultivated with a Triatoma infestans embryo cell line. Trypanosoma cruzi strain Peru was cultivated in the presence of an established cell line of Triatoma infestans embryo cells (TI- 32). Bloodstream trypomastigotes differentiated into amastigote-like cells (first differentiation phase) which multiplied to form large clusters of cells. Because of their clustering nature, a new term, "staphylomastigotes," has been proposed for this stage. After 10 days of cultivation, 90% of the staphylomastigotes underwent differentiation (2nd differentiation phase) to trypomastigotes (approximately 98%) or epimastigotes (approximately 2%). Bloodstream trypomastigotes cultivated without TI-32 cells underwent the first, but not the 2nd differentiation phase, although occasional epimastogotes were seen (less than 1%). The evidence presented suggests that TI-32 cells produce a labile factor(s) important not only for initiation of the 2nd differentiation phase but also for maintaining the parasites in the trypomastigote stage. The pH of the culture medium was not the initiating factor for the 2nd differentiation phase. Infectivity studies indicated that staphylomastigotes were as infective as blood stream trypomastigotes, but that metacyclic trypomastigotes isolated from culture after the 2nd differentiation phase were slightly more infective than bloodstream forms. Electromicrographs of styphylomastigotes do not provide any evidence of exchange of genetic material between cells.


Secretion of hexosaminidase isozymes by Tetrahymena. Tetrahymena pyriformis strain HSM secrete large quantities of lysosomal acid hydrolases into the medium. The finding that 2 isozymes of beta-N-acetylhexosaminidase (2-acetamido-2-deoxy-beta-D-glucoside acetamidodeoxyglucohydrolase; EC 3.2.1.30) could be resolved by DEAE ion exchange chromatography and of possible differences between the secreted mixture and the intralysosomal hexosaminidase activity suggested that Tetrahymena might prove useful for studies of the control of lysosomal hydrolase isozyme secretion. In the present paper, we report a considerable purification of these isozymes and describe a number of their kinetic properties. Four isozymes were isolated into 2 major forms, A1 and B1, and 2 minor forms, A2 and B2, which were similar to the respective major forms in all kinetic properties tested. Hexosaminidase B1 has a molecular weight of approximately 93,000 daltons and is inhibited by high concentrations of p-nitrophenyl-N-acetyl-beta-D-glucosaminide. The inhibition is reversed by ethanol. Hexosaminidase B1 has a molecular weight of approximately 93,000 daltons and is inhibited by high concentrations of p-nitrophenyl-N-acetyl-beta-D-glucosaminide. The inhibition is reversed by ethanol. Hexosaminidase A1 has a molecular weight of approximately 170,000 and is not inhibited by high concentrations of substrate. The A forms are relatively less active against p-nitrophenyl-N-acetyl-beta-D-galactosaminide than the B forms. Neither hexosaminidases A1 or B1 has any endo-beta-N-acetylhexosaminidase activity. Comparison of the properties of the 2 major isozymes suggested that measurements of activity obtained under different assay conditions could be used to quantitate the amount of each isozyme in a mixture of the two. Log- and early stationary-phase cells secrete approximately 20% of isozyme A and 80% of isozyme B into the medium or into a dilute salt solution. With increasing culture age the fraction of isozyme A secreted rises to over 90%. Supplementation of the proteose-peptone growth medium with glucose causes a decrease in total hexosaminidase subsequently secreted but with no change in proportion of each isozyme. Cells suspended in a dilute salt solution containing 0.1 mM L-propranolol secrete slightly more isozyme A than do control cells suspended without L-propranolol. Phenoxybenzamine (0.2 mM) causes a slight decrease in the proportion of isozyme A released.


Effect of structure on phenothiazine cation radical reactions in aqueous buffers. The reactions of the cation radicals of 11 phenothiazine tranquilizers were examined in mildly acidic aqueous buffers. Of the 11, those having an aminopropyl side chain in the 10 position reacted to form 0.5 mol of sulfoxide and 0.5 mol of parent drug per mole of initial radical. Cation radicals with different side chains react to form additional products, which remain to be identified but probably result from hydroxylation of the phenothiazine ring. The decay kinetics of three of the cation radicals undergoing reactions with known stoichiometry, namely, chloropromazine, promazine, and triflupromazine, were studied in detail, and it was concluded that they all react via the same mechanism. The mechanism involves attack of the cation radical by a nucleophile, and radicals with electron-withdrawing groups in the 2 position react more quickly. Since the cation radicals with faster reaction rates with nucleophiles are more pharmacologically active, it is hypothesized that the cation radical-nucleophile interaction may be responsible for binding of phenothiazines to receptor proteins.


In vitro studies on adenomatosis of the colon and rectum. Cell culture studies on adenomatosis of the colon and rectum (ACR) suggested that the clinical phenotype, colonic adenomas which become malignant recognised as a single clinical entity, may not be entirely the result of a single dominant mutation. Of the dermal cultures containing both epithelioid and fibroblastic cells established from six affected subjects from six unrelated ACR families, four showed increased tetraploidy and two did not. Of similar cultures established from four affected subjects form families with ACR associated with sebaceous cysts in consecutive generations, three did and one did not have increased tetraploidy. Irrespective of the in vivo relationship of increased tetraploidy to colonic cancer, the cultures from seven ACR patients had populations of tetraploid cells, at least in vitro, with chromosome instability. Such a difference in expression of the ACR genotype in vitro suggested genetic heterogeneity within this clinically defined group.


The normal microbial flora of the baboon vagina. The microbial flora of the upper vagina and cervix was examined in 38 adult baboons at various stages of the menstrual cycle. The mean number of different species isolated from each baboon was 9.5, with species of Bacteroides, Corynebacterium and group D streptococci predominating. Lactobacilli and mycoplasmas were found in 47.4 and 44.7% of the animals, respectively. No ureaplasms were isolated. Cyclical variations in the microbial flora were minimal.


The shape of red blood cells as a function of membrane potential and temperature. It is well known that a pH shift of the outside medium from 5 to 9 produces a shape transformation of washed human red blood cells from stomatocytes to echinocytes in isotonic salt solutions. In addition, a stomatocytogenic effect is demonstrated here due to solutions of low ionic strength (below 70 mM). An analysis of the true cell state in these situations, proved by measurements of predicted volume changes, indicates a good correlation between transmembrane potential and cell shape. The fact that amphotericin B acts as echinocytogenic agent in low ionic strength medium at pH 7.4 but not at pH 5.1 underlines this explanation. Therefore, a transmembrane potential positive inside produces stomatocytes, slightly negative inside (below--10 mV), normocytes, and strongly negative, echinocytes. The temperature dependence of this process underlines the rigidity-pattern hypothesis of red blood cell shape (Glaser & Leitmannová, 1975, 1977).


Studies on lithium transport across the red cell membrane. V. On the nature of the Na+-dependent Li+ countertransport system of mammalian erythrocytes. Ouabain-resistant Na+-Li+ countertransport was studied on erythrocytes of man, sheep, rabbit, and beef. A transport system, exchanging Li+ for Na+ in a ratio of 1:1, was present in all four species. Li+ uptake by the exchange system increased 30-fold in the order man less than HK-sheep less than LK-sheep less than rabbit less than LK-beef. This order is identical to that of ouabain-resistant Na+-Na+ exchange in these species, but bears no relation to the Na+-K+ pump activity. The activity of the Na+-Li+ exchange system varied up to 7 and 16-fold among individual red cell specimens from man and beef, the variability being much smaller in sheep and rabbit erythrocytes. The affinities of the system for Li+ and Na+ were similar among the species and individuals (half saturation of the external site at about 1 mM Li+ and 50 mM Na+, respectively). 50-60% of Na+-Li+ exchange was blocked by N-ethylmaleimide in all species. p-Chloromercuribenzene sulfonate inhibited the exchange only in beef and sheep erythrocytes (60-80%). The two SH-reagents act by decreasing the maximum activity of the system, whilst leaving its affinity for Li+ unaltered. Phloretin was a potent inhibitor in all species. 1 mM each of furosemide, ethacrynic acid, and quinidine induced only a slight inhibition. The Na+-Li+ exchange of human and beef erythrocytes increased 3.5-fold upon elevation of the extracellular pH from 6 to 8.5, the pH-dependence arising from a change in affinity of the system for the cations and being similar to that reported for ouabain-resistant Na+-Na+ exchange in beef erythrocytes. It is concluded that a transport system exists in the red cell membranes of the four species which can mediate ouabain-resistant exchange of either Na+ for Na+, Na+ for Li+, or Li+ for Li+. The exchange system exhibits essentially identical transport characteristics in the four species, but shows a marked inter- and intra-species variability in maximum transport capacity and some differences in susceptibility towards inhibitors. A similar transport system is probably present also in other tissues. The exchange system seems to be distinct from the conventional Na+-K+ pump and shows no clear relation to one of the furosemide-sensitive, ouabain-resistant Na+ transport systems described in the literature.


Sulfatide content and (Na+ + K+)-ATPase activity of skin and gill during larval development of the Chilean frog, Calyptocephalella caudiverbera. The sulfatide content, phospholipid concentration, and (Na+ + K+)-ATPase activity from skin and gills of different stages of larval development of Calyptocephalella caudiverbera (a Chilean frog) were analyzed. Additionally, the short-circuit current in skin was studied. When skin and gills, depending on the stage of larval development, present (Na+ + K+)-ATPase activity, they have a high ratio of sulfatide to amount of membrane and the phosphatidylserine concentration remains unchanged. Sulfatide content and (Na+ + K+)-ATPase activity in skin are in direct relationship with the level of sodium flux present during development. The specific enzymatic hydrolysis of sulfatide with partially purified arylsulfatase of pig kidney inhibits 100% of the ouabain-sensitive (Na+ + K+)-ATPase. The ouabain-insensitive ATPase remains virtually unchanged with the treatment, even with a high concentration of arylsulfatase or with ouabain present in the medium. These experiments strongly suggest a role of sulfatides in the (Na+ + K+)-ATPase activity and, as a consequence, in sodium ion transport.


Compositional relatedness of aldehyde reductases from several species. The amino acid compositions of several monomeric NADPH-dependent aldehyde reductases from a variety of species have been determined and analyzed by the difference index method of Metzger et al. (1968). The difference indexes among mammals range from 4.15 - 6.10 indicating considerable homology. Comparison of chicken aldehyde reductase with mammalian aldehyde reductases gave values in the range 6.8 - 9.9 suggesting a close relationship whereas the difference indexes for the enzymes from fruit fly and Baker's yeast versus vertebrate aldehyde reductases (10.9 - 14.4) indicate more distant relationships. The extent of sequence homology among aldehyde reductases from these species was estimated from a plot of difference index versus percent sequence difference for oxido-reductases of known sequence. From this plot, and using a mammal-chicken divergence time of 300 million years and a mammalian order split of 75 million years, the rate of evolution of aldehyde reductases was calculated to lie in the range 5.8 - 15.6% sequence difference per 100 million years. Comparison with rates of evolution of oligomeric dehydrogenases indicates that aldehyde reductases comprise the most rapidly evolving family of oxido-reductases. This is probably related to the monomericity of aldehyde reductases since there is a direct correlation between the number of subunits and the rate of evolution.


Effects of several beta-blocking agents on the development of hypertension in spontaneously hypertensive rats. Antihypertensive effects of chronic oral administration of adrenergic beta-blocking agents were assessed in SHR. Propranolol, pindolol, oxprenolol, atenolol and labetalol were used as beta-blockers and the effects of these compounds on the blood pressure and the heart rate were compared with those of hydralazine, a representative vasodilating antihypertensive agent. Propranolol, oxprenolol and atenolol produced a definite decrease in the heart rate; the development of hypertension was retarded. Pindolol produced antihypertensive effects only after a longer period of administration and such were associated with insignificant decrease in heart rate. With a shorter period of administration the drug produced only an insignificant fall of blood pressure with practically no change in the heart rate. With labetalol, a beta-blocker with alpha-blocking action, a fall of blood pressure appeared earlier and was of greater magnitude. Hydralazine produced a definite antihypertensive effect, which appeared immediately after administration and was associated with a tachycardia. In pithed rats, only pindolol produced a definite fall of blood pressure. On the basis of these findings, possible mechanisms of antihypertensive effects of beta-blockers were discussed.


The involvement of cytochrome P-488 and P-450 in NADH-dependent O-demethylation of p-nitroanisole in rat liver microsomes. These studies have shown that addition of p-nitroanisole to a reaction mixture containing rat liver microsomes resulted in an increase the reoxidation rate of NADH-reduced cytochrome b5. Fortification of rat liver microsomes with partially purified cytochrome b5 produces an increase in both NADPH-dependent and NADH-dependent p-nitroanisole O-demethylation activity. Antiserum to cytochrome P-450 isolated from phenobarbital-treated rat liver microsomes inhibited the NADH-dependent O-demethylation activity as well as the NADPH-dependent O-demethylation activity seen in rat liver microsomes. Addition of either purified cytochrome P-450 or cytochrome P-448 to an incubation mixture containing phenobarbital-treated rat liver microsomes enhanced the NADH-dependent p-nitroanisole O-demethylation activity. These results suggest that NADH-dependent and, in part, NADPH-dependent O-demethylations are catalyzed by cytochrome P-448 and cytochrome P-450 receiving electrons from cytochrome b5.


Stimulation by catecholamine of purine catabolism in rats and chickens. The effect of catecholamine in vivo was studied on purine catabolism in rats and chickens. Catecholamine, administered intraperitoneally in a high dose, markedly increased plasma uric acid and allantoin in rats, and an increase was also observed with intravenous infusion of a lower dose of catecholamine. The effects of catecholamine were characterized by inhibition with alpha and beta adrenoceptor antagonists. Regarding the mechanism of this catecholamine action on purine catabolism, it was shown that catecholamine stimulated degradation of tissue ATP into the end-product. Plasma allantoin, the final purine catabolite in rats, elicited by catecholamine could be maintained under conditions of renal failure, although the action of catecholamine in intact rat was short lasting. The effect of catecholamine was potentiated and/or prolonged by angiotensin-II and aminophylline, and a hyperuricemic state could be induced by catecholamine treatment in chickens. In addition, increase of plasma purine catabolite by immobilization stress in rats suggested the involvement of endogenous catecholamine. From these experimental results, it is considered that catecholamines probably play a important role in the pathogenesis of hyperuricemia.


Inhibitory alpha-adrenergic action of phenylephrine in guinea pig taenia caecum. Phenylephrine, a selective alpha-adrenergic stimulant, caused a maximal relaxation of the taenia from guinea pig caecum in the concentration of 10(-6) g/ml. Phenylephrine in this concentration did not influence intracellular cyclic AMP and cyclic GMP levels. Although phenylephrine abolished the spontaneous spike discharge, no change was detected in 45Ca-uptake and 45Ca-efflux on the tissue level after phenylephrine. Ca-uptake and Ca-release on the subcellular level were also not influenced by phenylephrine. In Ca free-solution phenylephrine inhibited the response to CaCl2. Phenylephrine increased 42K-efflux in the normally polarized taenia and also in the K-depolarized taenia.