Terbutaline in the treatment of acute asthma in childhood. Forty-eight children with known asthma (ranging in age from 2 to 16 years) were studied during an acute attack. Each received either terbutaline or epinephrine subcutaneously in a random double-blind fashion. Measurement of heart rate, respiratory rate, and systemic arterial systolic and diastolic blood pressures and careful clinical assessment of obstruction of the airway were made before and at 15, 30, and 60 minutes after the administration of the drugs. Appreciable and significant clinical improvement was noted in 19 of the 24 patients in both groups and was of comparable magnitude. A small, but significant, increase in heart rate was noted in those patients requiring only one injection of terbutaline, suggesting that the drug's selectivity for the lung is relative not absolute. The present study demonstrates that terbutaline is an effective bronchodilator drug in acute childhood asthma.


[20 years in the management of reflux disease using fundoplication]. During the past twenty years fundoplication has found widespread propagation. Due to its effectiveness it is today the most often applied operative procedure of reflux-preventing surgery. As such the method must be reserved for clearly demonstrated reflux disease and its organic complications. False indication and incorrect technique have burdened results of such operations with unnecessary complications. The steps to a successful surgical reconstruction of the cardia are thus shown. In the future, the results of this surgical method will have to be improved more by correct indication instead of changing the surgical technique.


[The telescope phenomenon. A complication possibility following fundoplication]. On the basis of four cases from our own clinic and three outside cases, we describe a so-called, "telescope phenomenon". This involves a specific complication that may arise after fundoplication. The characteristic of this postoperative complication is the slipping down of the fundus cuff, accompanied by a telescopic pushing upwards of the cardia. Recurrent reflux and occasionally new manifestations of dysphagia are the clinical results of this. This paper describes the diagnostic procedure and therapy of the telescope phenomenon.


[Results of esophago-intestinal anastomoses and studies of gastric, perfusion in the stomach mobilized for that purpose]. The results of 61 resections of the esophagus are reported. The cause of leakage is investigated. Using angiography and the TAS method, increasing diminution of the blood supply of the gastric wall can be observed after the ligature of 1, 2, or 3 arteries. Preservation of the right gastric artery and the right gastroepiploic artery, leaving an intact arcade, avoids necrosis of the suture line. However, it is better to remove the fundus in every case. Results after esophagointestinal anastomoses and investigations of blood flow in the mobilized stomach are reported.


Leiomyomas of the rectum and anal canal: report of six cases and review of the literature. Since 1872, 168 rectal leiomyomas have been reported in the world literature. Their incidence, therefore, is very low. It is estimated that one leiomyoma may be seen for every 2,000 or more rectal tumors. Those leiomyomas located in the anal canal and sphincter are the rarest. A case is reported here that appears to be the eleventh in such a location. The incidence of these lesions is highest between the ages of 40 and 59 years. Most patients have a combination of symptoms rather than a single one, the most frequent of these being the presence of a mass, bleeding, and constipation. Almost always, the chain of events that leads to the diagnosis starts when the tumor is discovered by digital examination of the rectum. The lack of reliable criteria of malignancy and the marked tendency to recur shown by these tumors are the two main problems the surgeon faces when selecting the operative procedure to be used. We believe that those tumors with an original size of 5 cm or more in largest diameter are the ones that have shown the highest tendency to recur, mostly as sarcomas. Therefore it is thought that these lesions should be treated radically from the beginning, especially when they recur. More adequate follow-up studies are needed.


Psychiatric aspects of allogeneic bone marrow transplantation for aplastic anemia. Observations attendant to daily psychiatric intervention with a patient undergoing allogeneic bone marrow transplantation are presented in a case report. The coping responses of the patient are delineated and discussed together with comments and suggestions regarding management.


[Islet-cell antibodies in juvenile diabetes mellitus (author's transl)]. Islet-cell antibodies were detected in 11 of the 67 patients with juvenile diabetes mellitus. These antibodies reacted with all endocrine-islet cells, although higher serum dilutions and different staining intensity of the various islet cells were noted. Antibody formation to islet antigens was found to be closely associated with HLA B8 (P = 0.03). However, there was no relationship between islet-cell antibody production and insulin antibody formation. The demonstration of pancreatic islet-cell antibodies, particularly in HLA B8-positive juvenile diabetics, constitutes further circumstantial evidence of a genetically determined auto-immune pathogenesis in some patients with juvenile diabetes mellitus.


[Prevention of postoperative thrombo-embolism by heparin/dihydroergotamine (author's transl]. Using the radiofibrinogen test and perfusion lung scanning (2-phase pharmacoscintigraphy), the incidence of postoperative thrombosis and pulmonary embolism was determined in a group of 362 operated patients. Low-dose heparin administration (twice 5000 IU daily) was given to 162, combined heparin and dihydroergotamine administration (5000 IU heparin and 0.5 mg dihydroergotamine daily) to 150, while 50 patients received no prophylactic treatment. There was a significant decrease in thrombo-embolism in the heparin/DHE group (8.7% deep-vein thrombosis; 2.7% pulmonary embolism) compared with the heparin group (19.8% and 5.5%, respectively) and the control group (30% and 14%, respectively).


[Insulin treatment of decompensated diabetes mellitus with a new artificial endocrine pancreas (author's transl)]. During the past decades insulin has been given in relatively high doses when treating diabetic coma. Recently low-dose insulin treatment has been proposed by several groups. In the reported investigation insulin was initially given in moderate to high doses (12-200 U/h) with a steady reduction in dose during the course of treatment. Insulin infusion was regulated either manually with an adjustable infusion pump (7 patients) or automatically with an artificial endocrine pancreas (glucose-controlled insulin infusion system; 11 patients). Thus 18 patients with decompensated diabetes mellitus (coma or precoma) were treated. In 14 patients with ketoacidotic decompensation laboratory data on hospital admission were: blood glucose 7.35 +/- 0.61 g/l, serum potassium 4.7 +/- 0.4 mmol/l, pH 7.1 +/- 0.04, base excess - 19,7 +/- 2.2 mmol/l (x +/- SEM). The other patients had hyperglycaemic or hyperosmolar non-ketotic decompensation. In all patients controlled reduction of blood glucose levels was achieved within 2.3 to 18 hours. The amounts of insulin infused during this ranged from 17 to 320 units, but in one instance was 1950 units. There were no complications.


[Polychemotherapy in non-hodgkin lymphoma with a modified MOPP scheme (author's transl)]. 64 patients with extensive non-Hodgkin lymphomas were treated with a modified MOPP scheme. 41 patients had stage IV disease. 59 patients responded to treatment. In 22 cases complete remission was obtained and in 37 cases partial remission. The proportion of complete remissions was the same for reticulosarcoma (9 out of 28) and lymphosarcoma (7 out of 22). The remission rate for Brill-Symmers disease was higher (6 out of 14). For patients with lymphosarcoma the average duration of complete remission was 26.9 months and for patients with reticulosarcoma 25 months. 42 of the 59 patients who responded to treatment survived one year, 33 of them two or more years. After a two-year period there were no more deaths.


[The behavior of the Q-fever agent, Coxiella burnetii, in birds. 3. Experimental infection of quail]. After simultaneous aerogenic and alimentary infection of quail by intra-nasal inoculation of a suspension of C. burnetii, the agent was reisolated 6 h after infection from lung and gut, from the 3rd day on from the spleen, at the 8th and 10th day from blood, and from the 8th day on from liver and kidney. C.burnetii was found in various organs up to 21 days after infection. In the majority of birds agglutinating antibodies could be demonstrated from the 18th day up to termination of the experiment 66 days after infection. On the basis of these results the course of a C. burnetii infection in birds is discussed.


[The arterial vessels and their intraosseous course in the leg bones of foals. II. Humerus]. The arterial blood vessels in 24 humeri of equine fetusses and foals are described. The relation between the age and the distribution of these arteries are explained and completely discussed.


[Phantoms for the collection of genital secretions in stallions]. Practical experiences of the phantom method for collection of genital secretions from stallions are reported. Taking a phantom used in the Richard-Götze-Haus Tierärztliche Hochschule Hannover as a prototype two further models slightly modified have been constructed, baring a flat hollow in the right side of the caudal phantom body for manual inserting of the Artificial Vagina. These three models fulfill four important conditions for routine use: (1) sufficient sexual attractivity for the stallions; 80-85% successful collections of presecretions out of a total of 1050 using the dummy and 70% successful semen collections from more than 240 in total; (2) solid and resistant construction; (3) easy cleaning and desinfection of the surface of the phantom to get representative samples; (4) firm installation on a hygienic floor.


[The behavior of calves under various housing conditions]. Investigations were performed regarding the length of rest, sucking and feeding and the daily rhythm of these activities in calves kept with their mothers on pasture or in stables and the motility of the calves. Length of rest and feeding in breeding calves kept in groups and length of rest and sucking in breeding and fattening calves (automatic feeding and bucket feeding) were examined too. The examinations were performed starting from the morning feeding to the end of the day over a period of appr. 10.5 to 12 hours. During this time, calves 2--4 weeks of age ran about 550,7 m, 6--8 weeks of age 423.7 and 10--12 weeks of age 388.1 m. The length of sucking under automatic feeding conditions was comparable to the datas obtained from calves kept with their mothers. Bucket feeding took 2--4 minutes regardless whether rubber teats were used or not. Fattening calves in groups or single showed feint sucking or licking activities of 70.0 and 114.0 minutes/day respectively, breeding calves 39.8 and 20.8 minutes/day resp. Feint sucking in connection with a deficit of real sucking was observed regularly under bucket feeding conditions but only in those cases of automatic feeding, where the calves pushed each other aside from the feeding facilities. Feint rumination was observed in all fattening calves when kept strawlessly and lasted for about 31 minutes/day. Neither the feint rumination nor the feint sucking ever reached physiological lengths. Both behavioural patterns may be important in regard to animal welfare considerations of special maintenance conditions.


[Electromechanical registration of the resting behavior of fattening pigs]. The resting behaviour of four weanling pigs has been continuously recorded by an electromechanical apparatus for 8 weeks. The duration of different postures: standing, ventral recumbency, lateral recumbency and frequency of standing periods were recorded as influenced by different environmental factors. 1. Floor with and without bedding at 21 degrees C. 2. Floor without bedding at 27 degrees C environmental temperature. 3. Feeding once daily versus twice. Duration of recumbency periods was increases at 21 degrees C if bedding was provided. This also improved daily gain. At elevated environmental temperatures the animals preferred the lying posture on concrete floor. Feeding twice increased the duration of recumbency. The same was the case as the animals grew older. Disturbance by caretaking activities in neighbouring dens increased the duration of standing.


[Specimen collection in phosphoric acid ester poisonings]. For investigation of intoxications with organic phosphates contents of stomach and intestines are the best analytical material. Phosphates are destroyed by metabolism in liver, so liver tissue should not be used as proof for organic phosphates.


[Reaction of the periodontium to the wire ligature splint (Stout-Obwegeser)]. The direct traumatising effect of the Stout-Obwegeser wire ligature splint on the marginal periodontium is minimal and need not be considered. The increase in depth of pockets, in the index of sulcus bleeding and in tooth mobility is due to poor oral hygiene. In 16 patients (orthodontic operation and jaw fractures) periodontal measurements were obtained on 369 teeth in 30 jaws before application of the splint, 1 and 6 weeks later and as 6 weeks after removal of the splint. All gingival indices determined were at first raised. The last measurements however showed, better values than those at the beginning of treatment. Inflammatory changes regressed near all teeth whether they were included in a ligature or not. This appears to prove that oral hygiene alone is decisive. The question whether the "curettage" effect of splint application has a therapeutic action is left open.


Development of an enzyme-linked receptor assay (ERA) for hCG. An enzyme-linked receptor assay (ERA) for hCG was developed using horseradish peroxidase. The 1,500 g pellets, interstitial cell fraction, and solubilized homogenate from the rat testis linked to tanned sheep red blood cells (SRBC) were used as the binding fractions for hCG. In these ERA systems, the limit of detection for hCG was 30 IU/l (3-12 mIU/tube), which was almost equal to that of radio receptor assay (RRA). The ERA using SRBC linked solubilized receptor fraction showed the most satisfactory result in accuracy, reproducibility and easiness.


Effects of morphine and naloxone on serum LH, FSH and prolactin levels and on hypothalamic content of LH-RF in proestrous rats. Proestrus surges of serum LH, FSH and prolactin (PRL) were significantly reduced when morphine HCl (50 and 10 mg/kg) was administered to 4-day cycling rats just prior to the proestrous critical period. The inhibitory effect of morphine was reversed by naloxone, a morphine antagonist, at the dose which had no effect on the proestrus surges of serum LH, FSH or PRL. The hypothalamic LH-RF content of proestrous rats at 1800 hr (during the proestrus surge) was not significantly different from that at 1400 hr (before the surge) and was not affected by pretreatment with morphine or naloxone. Our results suggest that naloxone reverses the anti-ovulatory effect of morphine by antagonizing the inhibitory effect of morphine on preovulatory surges of gonadotropins or PRL.


Effect of neonatal castration on the content of hypothalamic LHRH, pituitary LH and plasma LH in developing male rats. The content of hypothalamic LHRH and concentration of LH in pituitary and plasma were measured on day 5, 7, 10, 14, 17, 22, 25, 30, 45, 52 and 60 in male rats which were bilaterally castrated on day 2. The levels of plasma LH were significantly higher in all the groups of castrated rats than in normal male rats of corresponding ages. The concentration of plasma LH did not rise progressively but showed day to day fluctuation apparently due to alteration of sexual differentiation of the hypothalamus. The concentration of pituitary LH was significantly lower in neonatally castrated rats compared to normal male rats except on days 17, 25 and 30. The content of hypothalamic LHRH declined initially following castration, but from day 17 onwards significantly higher levels of hypothalamic LHRH were maintained in neonatally castrated rats than in intact control. Initial decline in the content of hypothalamic LHRH may be because of stimulation of release of LHRH which exceeds maximal rate of synthesis and subsequent increase in the content of hypothalamic LHRH may be due to enhanced LHRH synthesis as a result of castration.


Ontogeny of the hypothalamo-neurohypophysial system in rats--an immunohistochemical study. In fetal rats neurophysin has been visualized immunohistochemically first at the 16th gestation day in perikaryons of the supraoptic nucleus, followed by the median eminence and the neurohypophysis at the 17th day, and the paraventricular neurons at the 19th day. The external zone of the median eminence contains abundantly immunoreactive fibres at the first days post partum. In the perikaryons of the suprachiasmatic nucleus immunoreactive material appears after the 3rd day of postnatal development.


Studies on the (pro) insulin biosynthesis of islets of Langerhans of sand rats (Psammomys obesus). By feeding a regular laboratory chow, sand rats (Psammomys obesus) from our breeding colony gained different body weights, though they received approximately the same quantity of calories. Sand rats, reaching a body weight above 160 g (group B) showed significantly increased blood glucose values in contrast to the animals with a body weight under 160 g (group A). Isolated pancreatic islets of these two groups of sand rats were incubated with [3H]-leucine to study the incorporation of this amino acid into proinsulin and insulin. The incorporation into proteins of pancreatic islets of sand rats of group B was stimulated by 0.45 mg and 3.0 mg/ml glucose. In group A there was no further stimulation from 0.45 mg to 3.0 mg/ml glucose. Insulin secretion could be stimulated by glucose in both groups, but the stimulation was stronger in group B than in group A.


[Various disturbances in beta cell function in diabetes mellitus]. Various types of insulin secretion may be differentiated in diabetes mellitus. In juvenile diabetes, there can not be induced in general any stimulation of insulin secretion, whereas in maturity-onset type diabetes a significant increase of the serum level was found after stimulation with glucagon or tolbutamide. However, neither moderate glucose doses given orally nor very high glucose doses given intravenously were able to stimulate insulin secretion in some of these patients. These findings suggest a complete insufficiency of beta cell glucose receptors in this specific type of maturity-onset diabetes.


Occurrence of biogenic amines in the pancreatic islets of sand rats (Psammomys obesus). The islets of Langerhans of sand rats (Psammomys obesus) have been investigated ultrahistochemically in order to demonstrate biogenic amines. It could be found that amine-containing vesicles with a diameter of 600-700 nm from the type of catecholamine-granules exist in A-cells as well as in B-cells. It can be stated that the sand rat belongs to the species containing amines in A- and B-cells. The importance of the amines for the regulation of the hormone secretion of the islets of Langerhans is discussed and the connections to diabetes mellitus are referred too.


Electron microscopy analysis of the interaction between Escherichia coli DNA-dependent RNA polymerase and the replicative form of phage fd DNA. 1. Mapping of the binding sites. The interaction of Escherichia coli DNA-dependent RNA polymerase (EC 2.7.7.6) with the replicative form of the DNA from the filamentous coliphage fd cleaved by the restriction endonuclease HindII has been studied by electron microscopy at low and high ionic strength. In the presence of ATP or GTP, and heparin, RNA polymerase binds to fd replicative-form DNA at a few specific sites which have been mapped. The map was oriented so that transcription is from right to left. Three main GTP initiator sites are found at 15%, 82% and 94% of the genome length. One main ATP initiator site is found which cannot be mapped with the same accuracy, and which is localized between 38% and 50%. In the absence of initiator triphosphates and heparin, the binding of the enzyme to fd DNA is much more heterogeneous and therefore the mapping is more difficult. Nevertheless it seems that the preferential binding regions correspond to the specific sites mapped in the presence of GTP or ATP. The mean number of polymerase molecules bound to DNA as a function of the molecular ratio enzyme to DNA present in the mixture has been determined. From these results a binding isotherm can be obtained. The apparent equilibrium constant (K approximately 10(9) M-1) which is derived certainly represents an under-estimated value, as discussed.


Electron microscopy analysis of the interaction between Escherichia coli DNA-dependent RNA polymerase and the replicative form of phage fd DNA. 2. Analysis of the dissociation kinetics. The kinetics of dissociation of the fd DNA - RNA-polymerase complex has been analyzed. Heparin was added to a solution of the enzyme - DNA complex in order to trap free polymerases. At different times after, samples were taken and analyzed by electron microscopy to determine the mean number of enzymes bound per DNA molecule. Unexpectedly, the measured dissociation is not a first-order reaction. The apparent rate constant increases with heparin concentration in the range between 0.001 and 2 mg/ml. These results strongly suggest the existence of a direct transfer process of RNA polymerase to heparin, bypassing the rate-limiting step of dissociation of the enzyme - DNA complex to free enzyme. Theoretical analysis of the direct-transfer model shows that the rate constant of dissociation should level off at high heparin concentrations: measurements of the residual transcription activity show that this is the case. From these experiments, the equilibrium constant of the DNA - RNA-polymerase complex can be determined. The value K = 10(12) M-1 which is obtained solves a striking paradox which existed because measurements performed in other laboratories indicated K = 10(14) M-1, which is greater than the equilibrium constant of the lac-repressor - lac-operator complex (=10(13) M-1).


Modification of L-isoleucyl-tRNA synthetase with L-isoleucyl-bromomethyl ketone. The effect of the catalytic steps. The rapidly reacting cysteine-sulfhydryl group of L-isoleucyl-tRNA synthetase has been specifically alkylated with L-isoleucyl-bromomethyl ketone [Rainey, P., Holler, E. & Kula, M.-R. (1976) Eur. J. Biochem. 63, 419-426]. We have now investigated the catalytic and substrate binding properties of the modified protein by radioactive and fluorescence techniques. The rate constants for the transfer of AMP and isoleucine from the protein - adenylate complex to form ATP or Ile-tRNAIle were only 3% of those for native enzyme, whereas the rate constant for the formation of adenylate was essentially unchanged. The tendency to form synthetase - substrate complexes remained almost unchanged with the exception of L-isoleucine which exhibited a 20-fold reduction. Similarly, complex formation of L-isoleucinol together with its synergistic coupling to complex formation of ATP was partially inhibited. The results rule out the essential participation of the rapidly alkylatable cysteine-sulfhydryl group during catalysis.


Functional roles of 50-S ribosomal proteins. Ribosomal proteins previously inactivated by treatment with fluorescein isothiocyanate have been incorporated into 50-S ribosomal subunits during reconstitution from particles disassembled by 2 M LiCl in the presence of an excess of the modified proteins. The reconstituted particles show alterations in some functional activities resulting from the incorporation of the inactive ribosomal proteins added exogenously. Of the fluorescein-isothiocyanate-treated proteins incorporated, L24 and L25 drastically affect all the activities tested and these proteins possibly play a fundamental role in determining the overall structure of the particle. Proteins L16 and L10 are apparently involved both in the GTP hydrolysis dependent on elongation factor G and in peptidyl transferase activity but the modified protein L11 only affects GTPase activity indirectly and interferes with the ribosome assembly process involving proteins L7 and L12. Protein L1 may be involved with peptidyl transferase activity while proteins L7 and L12, in agreement with many reports in the literature, affect the factor-dependent hydrolysis of GTP.


Specificity of elongation factor Tu from Escherichia coli with respect to attachment to the amino acid to the 2' or 3'-hydroxyl group of the terminal adenosine of tRNA. Modified Tyr-tRNATyr and Phe-tRNAPhe species from yeast having the aminoacyl residue bound specifically to the 2' and 3' position of the terminal adenosine, respectively, were investigated for their ability to form ternary complexes with Escherichia coli elongation factor Tu and GTP. Both Tyr-tRNATyr-CpCpA (2'd) and Tyr-tRNATyr-CpCpA(3' d) derivatives which are esterified with the amino acid on the 3' and 2' position respectively and which lack the vicinal hydroxyl were able to form ternary complexes. The stability of these ternary complexes was lower than in the case of native Tyr-tRNATyr-CpCpA. Tyr-tRNATyr-CpCpA(3' d) having the amino acid attached to the 2' position interacted considerably more strongly with EF-Tu - GTP than Tyr-tRNATyr-CpCpA(2' d). Ternary complex formation was observed with neither Phe-tRNAPhe-CpCpA(2'NH2) nor Phe-tRNAPhe-CpCpA(3'NH2). It is concluded that 2' as well as 3' isomers of native aminoacyl-tRNA can be utilized for ternary complex formation but in a following step a uniform 2'-aminoacyl-tRNA - EF-Tu - GTP complex is formed. Although the free vicinal hydroxyl group of the terminal adenosine is not absolutely required, replacement of the ester linkage through with the amino acid is attached to tRNA by an amide linkage leads to loss of ability to interact with elongation factor Tu.


Affinity chromatography on agarose-hexyl-adenosine-5'-phosphate of methionyl-tRNA synthetase from Escherichia coli. Application of the couplings between the methionine and ATP sites. Recent studies by us [Biochemistry (1977) 16, 2570-2579] have shown that L-methioninol, a methionine analog lacking the carboxylate negative charge, enhances the affinity of AMP for methionyl-tRNA synthetase while L-methionine antagonizes the nucleotide binding. Such couplings between ligands of the enzyme have now been applied to affinity chromatography of methionyl-tRNA synthetase on an agarose-hexyl-adenosine-5'-phosphate gel (the spacer is attached to AMP at the adenine C-8 position). Retention of the enzyme on this gel column was shown to be dependent on the presence of appropriate concentrations of magnesium and of L-methioninol in the equilibration buffer. The enzyme was then specifically recovered from the column by omitting the amino alcohol or by adding an excess of L-methionine which antagonizes the cooperative effect of L-methioninol. This approach has provided the basis for a new purification procedure of methionyl-tRNA synthetase which leads to a 200-fold purification in a single chromatographic step. In this manner, after 30-50% ammonium sulfate fractionation of extracts of Escherichia coli EM 20031 (carrying the F32 episome), 0.25 mg X methionyl-tRNA synthetase was obtained at 90% purity per ml of agarose-hexyl-adenosine-5'-phosphate gel.


Somatic antigens of Shigella. The strucuture of the specific polysaccharide chain of Shigella dysenteriae type 5 lipopolysaccharide. The specific polysaccharide was released from Shigella dysenteriae type 5 lipopolysaccharide by mild acidic hydrolysis and then purified by gel chromatography on Sephadex G-50. The polysaccharide was built up of residues of D-mannose, 2-acetamido-2-deoxy-D-glucose, 3-0-(D-1-carboxyethyl)-L-rhamnose (rhamnolactylic acid) and 0-acetyl groups in a ratio 2:1:1:1. On the basis of radiospectroscopy, methylation analysis, Smith degradation, and chromium trioxide oxidation, the repeating oligosaccharide unit of the polysaccharide can be assigned the following structure: (formula: see text) where GlcNAc is 2-acetamido-2-deoxy-D-glucopyranose, Manp is mannopyranose, RhaLcA is rhammolacytic acid and Ac is an acetyl group. The serological properties of Sh. dysenteriae somatic antigens are discussed in relation to the chemical structures of their specific polysaccharides.


Ternary complex formation between elongation factor Tu, GTP and aminoacyl-tRNA: an equilibrium study. The equilibria between the elongation factor Tu-GTP complex (EF-Tu-GTP) from Escherichia coli and tyrosyl-tRNATyr from E. coli as well as phenylalanyl-tRNAPhe and seryl-tRNASer from yeast were studied using a novel procedure, which takes advantage of the protective effect of ternary complex formation on the stability of theaminoacyl bond against non-enzymatic hydrolysis. At 25 degrees C and at pH 7.4 tyrosyl-tRNATyr, phenylalanyl-tRNAPhe and seryl-tRNASer are bound with binding constants of 0.7 X 10(7) M-1, 5.0 X 10(7) M-1 and 0.5 X 10(7) M-1 respectively. The binding of aminoacyl-tRNA to EF-Tu-GTP has a negative deltaH of the order of 10 kcal/mol (42 kJ/mol). Complex formation is dependent on ionic strength: with 0.1 M KCl Kass = 0.8 X 10(7) M-1, with 0.5 M KCl Kass = 0.2 X 10(7) M-1 was determined for the binding of Tyr-tRNATyr.


Membrane proteins of Escherichia coli K-12: two-dimensional polyacrylamide gel electrophoresis of inner and outer membranes. Protein compositions of the inner and outer membranes of Escherichia coli K-12 have been analyzed by two-dimensional gel electrophoresis in which proteins are separated according to apparent isoelectric point (first dimension) and to apparent molecular weight (second dimension). Membrane proteins except for a pair of major outer membrane proteins (proteins Ia and Ib) were found to be solubilized effectively by lysis buffer containing urea, Triton X-100, ampholines and 2-mercaptoethanol. The latter two proteins could be solubilized after precipitation of membrane fraction with trichloroacetic acid; they formed a pair of spots at an acidic region on the electropherogram. Another major protein of the outer membrane, protein II, was also identified. Most of the inner and outer membrane proteins were shown to be focused at a pH range between 4 and 6.5. Specific protein patterns characteristic for both the inner and outer membranes could thous be visualized by the present system. At least 120 and 50 protein species were detected for the inner and outer membranes, respectively.


Structural fluctuation of the polypeptide-chain elongation factor Tu. A comparison of factors from Escherichia coli and Thermus thermophilus HB8. The kinetics of hydrogen-deuterium exhcange in the polypeptide chain elongation factor Tu (EF Tu) from Escherichia coli and that from Thermus thermophilus HB8 has been examined in aqueous solutions at various pH and temperatures by means of infrared absorption measurements. The free EF-Tu from E. Coli has a greater reaction rate at all pH values and at every temperature than that of the GTP-bound or GDP-bound EF-Tu. The free EF-Tu from T. thermophilus, on the other hand, has an alomst equal reaction rate to that of EF-Tu-GDP in the temperature range 38-55 degrees C. For the peptide NH groups belonging to a medium-labile kinetic class, a small but definite difference in the rate of exchange reaction was observed between EF-Tu-GDP and EF-Tu-GTP for both E. coli and T. thermophilus. For less labile peptide NH groups, on the other hand, the rate of the exchange reaction with EF-Tu-GDP from T. thermophilus is only slightly affected by the pH of the solution at 38 degrees C and 45 degrees C, while the rate constant(k) with E. coli EF-Tu-GDP is pH-dependent (log k oc pH). For T. thermophilus EF-Tu, heat stability measurements, kinetics of the rates of GDP and GTP dissociation, and circular dichroic measurements have also been made. The molecular basis for the thermostability of T. thermophilus EF-Tu is discussed.


Cell-wall lipopolysaccharide of the 'Shigella-like' Escherichia coli 058. Structure of the polysaccharide chain. Two lipopolysaccharide preparations were obtained from Escherichia coli 058 by extraction with 45% aqueous phenol and fractional precipitation with cetyltrimethyl ammonium bromide (Cetavlon). Chemical analysis and polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate showed that the two preparations differed only in the extent of the O-specific polysaccharide moiety. The O-specific polysaccharide was characterized with proton magnetic resonance and infrared spectroscopy, optical rotation and paper electrophoresis. Using gas-liquid chromatography and ion-exchange chromatography, it was shown to contain D-mannose, 2-acetamido-2-deoxy-D-glucose, 3-O-(R-1'-carboxyethyl)-L-rhamnose (rhamnolactylic acid), and O-acetyl groups in the molar ratios of 2:1:1:1. The polysaccharide and oligosaccharides obtained from it were subjected to methylation and chromic acid oxidation. The results obtained indicated that the polysaccharide consists of tetrasaccharide repeating units in which the trisaccharide beta-GlcNAc1 - 4alphaMan-1 - 4(2/3-O-Ac)-Man is substituted at C-3 of the non-acetylated mannose with rhamnolactylic acid. The repeating units are joined through alpha-mannosyl-1 - 3-glucosamine bonds. This structure is identical with that of the cell wall polysaccharide of Shigella dysenteriae type 5.


Reaction of yeast fatty acid synthetase with iodoacetamide. 3. Malonyl-coenzyme A decarboxylase as product of the reaction of fatty acid synthetase with iodoacetamide. Yeast fatty acid synthetase possesses very low malonyl-CoA decarboxylase activity. Treatment with iodoacetamide, while abolishing synthetase activity, induces a strong malonyl decarboxylase activity which, in turn, can be inhibited by N-ethylmaleimide. Kinetic analysis shows that the emergence of the decarboxylase activity is synchronized to the disappearance of the fatty-acid-synthesizing activity and thus, is due to carboxamidomethylation of the peripheral SH-groups of the multienzyme complex. Strong decarboxylase activity was also found after treatment of the synthetase with methylmalonyl-CoA. A hypothetical scheme is proposed which explains the origination of the decarboxylase activity as a consequence of conformational changes of the condensing enzyme component which happen when the peripheral SH-group is acylated or alkylated.


On the role of tryptophan in luteinizing-hormone-releasing hormone (luliberin). The tryptophan residue of luteinizing-hormone-releasing hormone (luliberin) was chemically modified to produce the following analogs: [Trp(o)3]luliberin, Trp-(2,4-dinitrophenylsulfenyl)-luliberin, Trp-(2-hydroxy-5-nitrobenzyl)luliberin, (Trp-S-luliberin)2, Trp-CH3S-luliberin and Trp-formyl-luliberin. The luteinizing-hormone-releasing activity of those analogs was determined by bioassay in vitro and found to be 0.2%, 0.2%, 0.6%, 1.5%, 1.7% and 7% of that of the natural hormone, respectively. These results demonstrate that alterations in the indole moiety of tryptophan-3, which lead to a reduction in its electron density or sterically restrict its electron avaliability, are associated with a dramatic loss of biological activity.


Investigation of the binding of Escherichia coli RNA polymerase to DNA from bacteriophages T2 and T7 by kinetic formaldehyde method and electron microscopy. The complexes of T2 DNA with RNA polymerase of Escherichia coli were studied by two methods: kinetic formaldehyde method with preliminary fixation of complexes with low formaldehyde concentrations, and electron microscopy. For electron-microscopic investigations the effect of different conditions of formaldehyde fixation for DNA-RNA-polymerase complexes was studied and optimal fixation conditions were found. The suggested fixation method for DNA-RNA-polymerase complexes allows investigation of RNA polymerase molecule distribution on DNA in a wide range of conditions (ionic strength of the solution, weight ration of enzyme to DNA etc.). The comparison of the concentration of RNA polymerase molecules bound to DNA, determined by electron microscopy, and the concentration of defects in DNA as determined by the kinetic formaldehyde method, showed their coincidence. The electron-microscopic procedure was used to make maps of RNA polymerase distribution on T7 DNA. A correlation between the binding regions of the enzyme and the genetic map of early DNA T7 region was found.


Small-angle X-ray titration study on the complex formation between 5-S RNA and the L18 protein of the Escherichia coli 50-S ribosome particle. The 5-S RNA (A) and the L18 protein (B) from Escherichia coli ribosomes form one single AB complex in the concentration ranges supposed to prevail in vivo; at concentrations of L18 higher than 40 mM there is some indication for a minor species, most probably an AB2 species. This is indicated from the X-ray scattering titration data of the 5-S RNA/L18 system recorded at 21 degrees C in ribosomal reconstitution buffer. As a result of the 1:1 complex formation, there is a relatively small but defined increase in the radius of gyration from 3.61 to 3.85 nm. This result as well as the experimental scattering curve can be explained by models where it is assumed that the elongated L18 model is quite far from the electron density centre and where protein L18 interacts with one or both of the minor arms of the supposed Y-shaped 5-S RNA molecule.


Group B streptococci: a new threat to the newborn. 8 newborns with early onset group B streptococcal infection and two patients with late onset meningitis were observed during a period of three years. Respiratory distress, early onset of apnoic spells, and roentgenographic signs of hyaline membrane disease or perinatal pneumonia may lead to early diagnosis, especially if shock develops. The fatal course can only be prevented by prompt antibiotic treatment.


Preliminary experience with extracorporeal renal surgery and autotransplantation. 31 patients with renovascular hypertension, calculous disease, short ureter or renal carcinoma (single kidney) were treated with renal autotransplantation, in 3 patients bilaterally. There were 2 deaths due to cardiac arrest and myocardial infarction. Secondary nephrectomy was performed in 2 patients, and thus 29 of 31 autotransplanted kidneys were salvaged.


Auxiliary liver transplantation in the rat, influence of the condition of the recipient's liver on the fate of the graft. To evaluate the influence of the functional state of the recipient's liver on the fate of an auxiliary liver graft in rats, diverse surgical interventions were carried out on the recipient's liver following transplantation of an auxiliary liver. All grafts consisted of 30% of the liver mass, were supplied with portal blood only, and provided with bile drainage. Permanent graft hypertrophy was observed when the recipient's liver was resected subtotally, in addition to the ligation of the bile duct. Ligation of the bile duct performed separately has a more pronounced effect on the increase of the weight of the graft than subtotal hepotectomy. If only portal blood was deviated to the graft, grafts atrophied. It is concluded that the functional efficiency and condition of the graft vary directly with the degree of functional impairment of the recipient's liver, compair functional hypertrophy. The present results do not seem to indicate the presence of a specific hepatotrophic factor in portal blood.


Supravesical urinary diversion in renal transplantation. Urinary diversion may be used in patients without a bladder or with irreversible, lower urinary tract abnormalities who might not otherwise be suitable candidates for renal transplantation. Three cases have been described to illustrate three different methods of supravesical urinary diversion that have been employed in association with renal transplantation.


An experimental model for extracorporeal renal surgery: ex situ removal of artificially induced renal staghorn calculi in ambiothermia. An experimental model for the evaluation of surgical techniques and preservation in extracorporeal surgery is described: staghorn calculi were induced in 15 dogs by the injection of methylmethacrylate into the renal pelvis. Removal of the calculi was carried out 3-4 weeks later on a workbench, using microsurgical techniques and ex situ radiographies. 15 min of warm ischaemic and 6 h of ambiothermic preservation were well tolerated as could be demonstrated by determination of the serum creatinine following autotransplantation and immediate contralateral nephrectomy.


Uptake of horseradish peroxidase by geniculo-cortical axons in the golden hamster: analysis by computer reconstruction. Micro-injections of horseradish peroxidase (HRP) were made into the visual cortex of the golden hamster. The "projection lines" of labelled neurons in the dorsal lateral geniculate nucleus (LGNd) were three-dimensionally reconstructed, using a computer graphics technique. The lines run rostrally and medially from their origins at the lateral surface of the nucleus. Using an anatomically determined retinotopic map of the LGNd, the positions of all labelled cells near the lateral surface were converted into equivalent visual field co-ordinates and displayed on a physiologically determined retinotopic map of the primary visual cortex. Comparison between the scatter of these equivalent retinotopic loci and an actual reconstruction of the injection site revealed that: 1. there was general agreement between the independent retinotopic maps of LGNd and visual cortex; 2. there was greater retinotopic scatter of labelled LGNd cells than could be accounted for by the area of tissue injury in the cortex; 3. the retinotopic scatter matched more closely the total visible halo of HRP staining in the grey matter; 4. HRP can be taken up from a cytoarchitectonic field into which it diffuses after injection into a neighbouring area; 5. HRP is probably not taken up by undamaged axons in the white matter. These results are compared with those obtained in other animals and other systems. No general rules emerge, but the possibility of uptake from wide areas of diffusion must be considered when interpreting results of HRP injection.


Renal biopsies performed on diabetics.

Renal biopsies performed on diabetics. Metabolic disorders and immunological factors are discussed in connection with the pathogenesis of diabetic microangiopathy. Renal biopsies were obtained from 22 diabetics (8 women aged 18 to 53, 14 men aged 15 to 52). 7 of the 22 patients had been suffering from diabetes for 2 weeks to 3 years, 10 for 7 to 25 years, 2 showed a pathological glucose-tolerance test, i.e., they had been "latent" diabetics, and 3 patients, had been so-called "potential" subjects of diabetes due to hereditary traits or delivery of big babies. They were examined by light miroscopy as well as by immunofluorescence microscopy. A number of cases were chosen for the differentiation and counting of glomerular cells (n=8) as well as for electron microscopic (n=7) and polarizing-microscopic (n=6) examinations. Histologically, focal proliferations of mesangial cells as well as an increase in mesangial substance in the glomeruli was found in all cases, although in a varying degree of intensity. These results were confirmed by both the glomuerular cell count and electron-microscopic examination. Immunofluorescence microscopy made it possible to detect frequently both IgA (9/17) and IgG (9/17), usually in either linear or mesangial arrangements whereas it was less frequently possible to detect IgM (1/17) and albumin (1/8) and impossible to detect beta1C in the glomerulus. Labeled insulin was detected five times in the glomerulus. Polarizing-microscopic measurements made in order to discover possible submicroscopic variations in the structure of GBM showed deviations in the average values of anisotropic indices from the controls in the group of long-term diabetics only. The pathogenesis of diabetic microangiopathy may be described as an inflow of immunoglobulins and serum proteins into the mesangium because of an alteration of the capillary endothelium, the mesangial cell being thus caused to overfunction, proliferate and produce an excess of mesangial matrix. In prolonged diabetes the mesangial cell, so far as its own metabolism is concerned, will finally be affected to the point where its power of synthesis is modified in the sense of an excess and/or faulty composition of GBM (glomerular basement membrane).


[Effects of somatostatin on pancreatic isolated islets and peripheral tissues of rats]. This study was performed in order to evaluate the effects of somatostatin on insulin releasing mechanisms and on glucose uptake in peripheral tissues using isolated pancreatic islets, isolated rat diaphragms and epididymal fat pads. Insulin release by various concentrations of glucose were examined, and it was found that 100 ng/ml of somatostatin significantly inhibited insulin release at the glucose concentration of 200 mg/dl. Somatostatin also significantly inhibted insulin release by the administration of 5microgram/ml of glucagon with 200 mg/dl of glucose concentration and 20 mM of orginine with 200mg/dl of glucose concentrations. But at the glucose concentration of 50mg/dl, no significant inhibition of somatostatin on insulin release was observed even when various concentrations of glucagon or arginine were added. The influence of somatostatin on peripheral tissues was examined in vitro, and no significant change on glucose uptake compared with the control group was shown in either tissues. The results indicated that somatostatin directly inhibited insulin release from rat pancreatic islets but had no effect on glucose uptake in peripheral tissues. The inhibitory effect of somatostatin on insulin release may act through the common mechanism of both glucose and other substances in leading to insulin release.


Warts in a pediatric renal transplant population. 18 of 49 pediatric renal transplant recipients, all of whom were on continual steroid and immunosuppressive treatment, were found to have warts. Although all 18 patients received treatment for this problem, only 5 had the warts successfully eradicated. The frequency of warts among transplant patients is three times that of the general childhood population and twice that of a group of nephrotic pediatric patients who received immunosuppressive therapy. We propose that drug-induced suppression of cell-mediated, and to a lesser extent, humoral immunity, may predispose people to develop warts.


Effects of glucose on the "in vitro" synthesis of ribonucleic acid in pancreatic islets from mice homozygous for the mutation "diabetes" (db). Isolated pancreatic islets obtained from spontaneously diabetic mice were recently shown to have a considerably reduced rate of insulin biosynthesis in response to glucose. This defect has now been further evaluated by in vitro measurements of the effects of glucose on the islet RNA metabolism. An increase of the glucose concentration from 3.3 mM to 16.7 mM more than doubled the incorporation of 3H-labelled uridine into the islet RNA in the normal mice. By contrast, no stimulation was observed in the diabetic mice. Electrophoretic separation, on polyacrylamide gels, of RNA from normal mouse islets indicated that glucose stimulated the incorporation mainly into RNA bigger than 4S. Furthermore the fraction of islet RNA which bound the poly (U) was stimulated in the normal mice but not in the diabetic animals. Taken together these findings indicate a deficient glucose regulation of the RNA metabolism in the diabetic mice which conforms to the previously reported low rate of insulin biosynthesis in response to glucose in these animals.


Hydroxyacyl CoA dehydrogenase, an enzyme important in fat metabolism in different cell types in the islets of Langerhans. The pancreatic islets show a remarkably high activity of L-3-hydroxy-acyl CoA dehydrogenase, an enzyme which operates in the fatty acid cycle by catalyzing the NAD+ oxidation of some of the degradation products. In order to study the distribution pattern of its activity within the islets, samples with different relative contents of A1-, A2- and B-cells were prepared and analyzed. The results show that it is unlikely that either the A1-cells or the enzymatically well equipped A2-cells contribute to the high activity values of the islets. In contrast, the experiments indicated that the high activity was due to the B-cells. After 72 hours starvation, leading to an increase in the serum free fatty acids, there was no change in the activity of the A2-cells, while the B-cells, however, showed a significant but moderate decrease in their activity. It is concluded that the B-cells are enzymatically equipped for the oxidation of fatty acid degradation products even in situations with diminished activity such as occurs during a decrease of the mitochondrial assembly.


Single nightly administration of ketoprofen (Orudis) and indomethacin suppositories in the management of osteoarthritis: a double-blind trial. In a double-blind trial carried out on 60 patients with osteoarthritis at various localizations, ketoprofen (Orudis), administered as a single nightly 100 mg suppository for ten days, was significantly more active and better tolerated than indomethacin, administered in the same way. Patient's and doctor's assessment was also in favour of Orudis. Laboratory examinations were not affected by either drug.


Tibicorten and betamethasone valerate: a double-blind comparative study. Tibicorten, a new fluorine-substituted corticosteroid for topical application, was compared with betamethasone valerate in a double-blind investigation with left-right examinations. There appeared to be no significant difference between the results obtained with each substance in the treatment of patients with acute or chronic eczema or with psoriasis.


Butacote: the first assessment in general practice. A double-blind crossover trial in the treatment of rheumatic conditions in general practice compared the effect and tolerance of 100 mg t.d.s. standard phenylbutazone (Butazolidine) with enteric-coated phenylbutazone (Butacote), each preparation being given for one week to fifty patients. There was no difference in effectiveness. Nine patients complained of dyspeptic symptoms, three during the first week, six during the second week, but none during treatment with Butacote. This, together with the significant 17:7 patient preference, clearly demonstrated the superiority of Butacote.


Butacote and ibuprofen: a comparative assessment in rheumatic diseases in general practice. A multicentre double-blind trial in general practice compared Butacote (enteric-coated phenylbutazone) 300 mg daily, ibuprofen 1200 mg daily, and a placebo in the treatment of rheumatic conditions. Each patient recieved two of the three treatments for one month each. Twenty-nine doctors admitted 193 patients. One hundred and sixty-eight patients (sixty-four with inflammatory polyarthritis, and sixty-three with osteoarthrosis) completed the study, which showed that Butacote was significantly better than both ibuprofen and placebo for the relief of pain and morning stiffness, and improvement of function. Butacote was significantly preferred to both ibuprofen and placebo by patients and doctors, to placebo by the patients. Ibuprofen was significantly better than placebo for relief of morning stiffness and for reducing the amount of supplementary analgesics. All three preparations were well tolerated and showed a similar incidence of gastric side-effects. It is concluded from this study that Butacote is more effective and as well tolerated as ibuprofen in the treatment of rheumatic conditions in general practice.


Butacote and naproxen: a comparison of effectiveness in rheumatoid arthritis. A double-blind crossover trial compared Butacote, 200 mg b.d. with naproxen, 250 mg b.d., each given for four weeks, in the treatment of rheumatoid arthritis. The trial was multicentre, and twenty-six general practitioners admitted forty-eight patients. Seven patients dropped out: two for technical reasons; one in each treatment group because of exacerbation of symptoms; one because of intolerance of rescue analgesic; and two with gastric intolerance to naproxen. Both drugs were effective in relieving pain, morning stiffness, and joint tenderness, as compared with the condition before the trial. There was little effect on grip strength or joint size, and no real differences between the two drugs. The two preparations had a similar frequency and spectrum of side-effects, except that oedema did not occur with naproxen, and rash did not occur with Butacote. Gastro-intestinal upsets were the commonest unwanted effect but only two patients had to stop treatment for this reason, both while taking naproxen. Although the doctors preferred each drug on an equal number of occasions, the patients preferred Butacote to naproxen almost twice as often (20:11), particularly when Butacote was given after naproxen. The reason for this divergence of opinions is not obvious.


A controlled comparative trial of Butacote and fenclofenac in the treatment of ankylosing spondylitis. A double-blind crossover trial of fenclofenac was untertaken to compare the short-term effects of phenylbutazone (Butacote) therapy with fenclofenac therapy (1200 mg daily) in 23 patients suffering from ankylosing spondylitis. The patients were randomly allocated to each of the test drugs for a period of two weeks, following wash-out periods of paracetamol for four days. The results favoured Butacote therapy in a dosage of 300 mg daily in the expressed preferences of both the patients and the physicians conducting the trial. Butacote was also better tolerated, as no severe side-effects occurred during treatment, whereas five patients developed a widespread rash during the period of fenclofenac therapy. No significant differences were seen between the two drugs in terms of clinical measurements of pain, morning stiffness, spinal movements, chest expansion, and abduction of the hips. Butacote was shown to cause some significant depletion of haemoglobin level, probably due to water retention, and Butacote also reduced the serum urate level to a significant degree.


System organization of alimentary behavior in the newborn and the developing cat. This review deals with the structural, behavioral, and EEG correlates of searching for food and food intake. Data were collected on freely behaving kittens of 1 to 60 days of age with chronically implanted electrodes. The 1st manifestations of hunger involve activation of limbic-reticular structures and electroencephalographic (EEG) activity consisting of fast oscillations of 30-40-60 cps, correlated with general behavioral arousal as manifested by motor restlessness and vocal reactions. The more specific behavior form of hunger--feeding motivation--involves structures that include the motor and parietal cortex, amygdala, and medial hypothalamus; the EEG activity consists of high-amplitude, slow waves of 3-6 or 6-8 cps; and the behavioral correlate is purposeful search for the mother's teat or food. Satiation is associated with activity of the synchronizing structures in the forebrain and lower brain stem, as well as in the relay nuclei that transmit the sensory flow from oral afferents; EEG activity consists of regular rhythms of 10-14 and 2-4 cps; and the behavioral correlate is the act of feeding.


In-vivo measurement of glucose and alanine metabolism with stable isotopic tracers. This report describes sensitive, precise microtechniques that allow quantification of glucose and alanine metabolism in vivo using stable isotope tracers. By combined gas chromatography--mass spectrometry and selected ion monitoring--deuterium and carbon-13 enrichment in blood glucose and alanine were measured with an error of less than 2 per cent. Estimation of glucose and alanine flux in dogs by stable isotope tracer techniques was compared with simultaneous measurements made in the same animal with conventional radiotracer dilution methods. Application of the described stable isotope methods to determination of glucose and alanine turnover as well as alanine-2,3-13C incorporation into glucose in adult men confirmed the safety and validity of these techniques for human investigation.


The effect of short-term glucagon infusion on kidney function in normal man. Kidney function was studied in six normal males before and during a 2 h glucagon (10 ng/kg/min) infusion. The following variables were determined during each 20 min clearance period; glomerular filtration rate (GFR), renal plasma-flow (RPF) , filtration fraction (FF), urinary albumin and beta2-microglobulin-excretion rates. Glucagon infusion resulted in a fourfold increase in plasma glucagon concentration. The infusion induced a significant increase in GFR (+9%), FF (+9%) and urinary beta2-microglobulin excretion rate (+32%), (p less than 0.01). RPF and urinary albumin excretion rates were not significantly changed. We suggest that glucagon may contribute to the reversible kidney function alterations typically found in poorly regulated juvenile diabetes, a state with relative or absolute hyperglucagonaemia.


Studies on translocation of immunoglobulins across intestinal epithelium. IV. Evidence for binding of IgA and IgM to secretory component in intestinal epithelium. We conducted studies concerning the issue of whether secretory component (SC) is a specific receptor on intestinal epithelial cells for IgA and IgM. Initially, frozen sections of human intestinal mucosa were incubated with dimeric monoclonal human IgA, conjugated to horseradish peroxidase; adjacent sections were reacted with peroxidase-conjugated antibodies to SC. The conjugated IgA and anti-SC bound to similar sites in the epithelium, that is to basolateral margins and supranuclear cytoplasm of columnar epithelial cells, principally in gland crypts. In subsequent tests of binding specificity, binding of the dimeric IgA conjugate was inhibited by pretreating the tissues with unconjugated dimeric IgA or 19S IgM, pretreating the tissues with unconjugated antibodies to SC, or preincubating the dimeric IgA conjugate with free SC. Binding was not inhibited or only partially inhibited by pretreating the tissues with monomeric IgA or IgG, pretreating the tissues with antibodies to human or heterologous immunoglobulins, or preincubating the dimeric IgA conjugate with 11S secretory colostral IgA. The findings indicate that dimeric IgA and 19S IgM are capable of binding in vitro to specific sites on intestinal epithelial cells, most likely to SC. This supports the hypothesis that transport of these immunoglobulins into intestinal fluids involves their combination with SC in the epithelium.


Rapid progression of chronic active type B hepatitis in a patient with hypogammaglobulinemia. Rapid progression of acute type B hepatitis to chronic active liver disease and cirrhosis in a young male with hypogammaglobulinemia is described. Absent circulating IgA, significantly low IgG, and normal IgM levels were detected during the acute phase of illness. Enumeration of peripheral lymphocytes revealed a decreased number of T cells and normal numbers of B cells. In vitro pokeweed stimulation of Ig synthesis correlated with the in vivo circulating levels of the three immunoglobulins. Cell-mediated immune responses were normal except for lymphocyte stimulation to hepatitis B surface antigen. It was concluded that the defective synthesis of IgG and IgA antibodies to hepatitis B surface antigen contributed to the accelerated progression to chronic active type B hepatitis in this person.


Analysis of the role of recombination and repair in mutagenesis of Escherichia coli by UV irradiation. Multiple mutant strains have been tested or their mimicry of the UV-mutagenesis deficiency of a recA single mutant. Revertants to histidine prototrophy and clear plaque mutants of lambda were scored to determine capacity for UV-mutagenesis. Nearly normal capacity was shown by a uvr+ recB- recF- strain, which shows almost no recA-dependent recombination, by uvr- recB+ recF- strains, which show almost no recA-dependent repair and by a uvrA- recB- recF- strain, which shows neither recA-dependent recombination nor repair. Since the uvr mutants can be assumed to show additionally no excision repair, these results may mean that UV-mutagenesis occurs during processes other than recombination and repair. Alternative hypotheses are discussed. The slight difference in mutagenic capacity was traced to the recF single mutation, which blocks the production of unmixed bursts of clear-plaque lambda mutants. Since this accounts for only about 10% of the mutations leading to clear-plaque mutants, it is suggested that there is more than one UV-mutagenic process.


[Multiple mutants of Saccharomyces cerevisiae. IV. Mutability of yeast cultures at different growth stages]. Mutability at different stages of culture growth in liquid media of two yeast strains, which revealed a property of "multiple mutability" and one strain of wild type for this property, was studied. One strain that possessed the property of "multiple mutability" showed at stationary phase a very high frequency of mutability, which reached 3.5%. It was found that multiple mutants arose in both strains, that possessed the property under investigation, at lag- and log-growth phases, and only in one strain -- at the stationary phase of growth. The possible reasons of "multiple mutability" display are discussed.


[Single-stranded conjugation in Escherichia coli K-12: characteristics of the formation of a heterogenous progeny]. During a single-stranded conjugation donor DNA, being a single-stranded form, takes part in the process of recombination. That is why a heteroduplex DNA must be an intermediate product of the recombination. The heteroduplex can be partially corrected as it was supposed for genetic transformation. The division of such a corrected heteroduplex gives the heterogenous progeny of exoconjugants. And this "correctional" heterogeneity must possess two following properties: 1) the mixed progeny must consist of only two recombinational genotypes; 2) the heterogeneity must be marker-specific. The experimental support to both predictions was obtained by the method of clonal analysis of conjugational merozygotes.


[Genetic control of Escherichia coli K-12 strains' assimilation of 2,6-diaminopurine as a purine source]. Mutations of the resistance to 2,6-diaminopurine (apt), which affect adenine phosphoribosyltransferase, fail to permit the growth of Escherichia coli pur mutants (purine auxotrophs which cannot make inosine monophosphate de novo) on the medium with 2,6-diaminopurine (DAP) as the sole source of purines. Addition of a small amount of hypoxantine, but not guanine, stimulated the growth of mutants of pur apt and pur apt+ genotypes on the medium with DAP. The utilization of DAP as purine source in the presence of hypoxantine is blocked by mutations guaC (guanosine monophosphate reductase), add (adenosine deaminase) and pup (purine necleoside phosphorylase), suggesting that DAP are utilized via purine nucleoside phosphorylase and adenosine deaminase. The drm mutation (that increases the level of pentose-1-phosphate in the cell) does not activate the utilization of DAP. The results indicate that a step, that limits the utilization of DAP as the sole source of purines by pur mutants of E. coli, is the deamination of DAP nucleoside.


[Escherichia coli cell competence induced by calcium cations]. Escherichia coli K-12 cells grown to early and late exponential, and early and late stationary phases were treated with CA2+ and analysed for the ability of exogenous 14C-DNA uptake and genetic transformation. DNA-membrane complexes were detected detected by isopicnic centrifugation of cell lysates in sucrose density gradient. It is found that 1) during all the tested phases of the growth cycle, E. coli cells attain the ability of enhanced DNA uptake after Ca2+ treatment; 2) exogenous and host DNAs are associated with the cell membrane during all tested growth phases; 3) nevertheless, the ability to form transformants is strongly time-dependent: the cells can be transformed during logarithmic phase only; 4) Ca2+ protects exogenous DNA against its degradation by bovine pancreatic DNAase. The peculiarities of Ca2+-induced competence, actual and possible interference of Ca2+ at different transformation steps are briefly discussed.


[Use of 8-methoxypsoralen and light (lambda--365 nm) for studying repair in yeasts]. The lethal effect of 8-metoxypsoralen (8/MOP) plus light (lambda = 365 nm) on the haploid radioresistant and UV-sensitive strains of Saccharomyces cerevisiae was studied. The mutation uvs1 increased the sensitivity to the lethal effect of 8-MOP more than 2.8 times as compared to radioresistant strain. The method of repeated irradiation allowed to study kinetics of excision of monoadducts induced by 8-MOP. The mutant uvs1 was characterized by the absence of excision of monoadducts. The radioresistant strain removed monoadducts very efficiently (80%) after the incubation in complete liquid medium for 2.5 hours at 28 degrees before repeated irradiation. After the incubation of this strain in buffer (pH 7.0) monoadducts were removed considerably less efficiently (30%).


[Site-specific recombination between phages lambda and phi 81 and integration of hybrid phages lambda-phi 81]. Dependence of the formation frequency of hybrid phages immlambdahphi81 and immphi81hlambda in recombination between phages lambda and phi81 from int-function of both phages is studied. Phages with hybrid att-sites (Plambda OP'phi81 and Pphi81OP'lambda) are isolated and the efficiency of integration of these phages into bacterial chromosome is determined.


[Salmonella typhimurium--test system for detecting the mutagenic activity of environmental pollutants. II. Mutagenic effect of heavy metal salts in an in vitro system with metabolic activation]. Mutagenic effect of zinc chloride on Salmonella typhimurium strain was detected using in vitro metabolic activation system. Cadmium chloride showed no significant mutagenic activity in the same system. It is recommended to use both in vitro and in vivo metabolic activation systems in mutagenicity testing of chemicals.


Influence of acetylsalicylic acid and paracetamol on menstrual blood loss. In a double-blind cross-over study the influence of acetylsalicylic acid (ASA) and paracetamol on menstrual blood loss has been investigated in 23 women without an intrauterine loop or birth control pills. After intake of ASA there was no significantly, increased blood loss compared with intake of placebo or paracetamol.


Dorsal silicone rods in the primary care of war injuries. During armed combat serious hand injuries are often neglected only to receive the attention they deserve far too late. The authors have employed a vigorous approach to the primary care of severe war injuries to dorsum of the hand introducing silicone rods where extensor tendons have been destroyed and where skin closure is obtained using inguinal skin flaps.


[Complications and failures through negative selection in replantation of amputated extremities]. Two years of experience with our replantation service in Vienna serves to define the criteria by which an amputated member is deemed worthy of replantation. 68 cases are analyzed and the complications and failures are discussed. The criteria took into account the type of injury, the ischemia time, the method adopted to preserve the severed part as well as the functional need of the amputee. We believe that by today's standard of replantation surgery, with the aid of microsurgical methods a responsible approach to indications would improve the functional results. Replantation capability, replantation possibility, replantation readiness and replantation obsession are all discussed.


[Complications in free flap transfer with microvascular anastomosis]. We used free flap transfers with microvascular anastomoses to cover defects on head, leg, foot and hand. In our experiences of 12 cases we got 2 total and 3 partial necroses of the transplants. The possible causes of the failures are discussed, and the results are compared with other reports. The significance of the method in hand surgery is demonstrated, comparing groin and dorsalis pedis flaps.


[The hypothalamo-neurohypophysial system and antidiuretic hormone (author's transl)]. Production, transport, storage and release of antidiuretic hormone (ADH) in the hypothalamo-neurohypophysial system were investigated. ADH produced by nerve cells in the paraventricular and supraoptic nuclei of the hypothalamus is present in a form bound to the specific protein neurophysin, in the neurosecretary granula. Electric and chemical stimulation of these nuclei results in evoked release of ADH in ionic association with neurophysin from the neural lobes. Acetylcholine, norepinephrine, histamine, angiotensin II, gamma-aminobutyric acid and L-glutamic acid have been regarded as candidates of chemical transmitters for the release of ADH in the hypothalamus. Prostaglandin (PG) E2 may be another important compound for central regulation of water metabolism. The possibility that PGE2 may be the transmitter or a modulator in the nuclei has to be considred. Serotonin, dopamine and taurine, however, may not be involded in the ADH releasing mechanisms in the hypothalamus. It appears that norepinephrine, histamine, angiotensin II, PGE2 and bradykinin stimulate directly the neural lobe to release ADH. The ADH release is regulated by intracellular Ca++. The existence of a "readily-releasable pool" of ADH can be ruled out and any limitation in the amount of ADH released under experimental conditions may be due to insufficient activation of the neural lobe. A physiological significance other than a carrier was proposed for neurophysin.


[A comparison of the effects of the beta-adrenergic blockers, carteolol (OPC-1085), propranolol and alprenolol on isolated rat cardiac muscles (author's transl)]. The effects of carteolol, propranolol and alprenolol were studied by using spontaneously contracting atria, atrial muscle and papillary muscle prepared from rat hearts. Propranolol, alprenolol and carteolol caused decreases in atrial rate and in contractile tension of electrically driven cardiac muscles in concentrations of 10(-6) to 10(-5) g/ml, 10(-6) to 10(-5) g/ml and 10(-4) g/ml, respectively. These compounds, also, effectively prevented electrically-induced atrial arrhythmias in these concentrations. Contractile tension of the cardiac muscles was increased by carteolol in concentrations of 10(-7) to 10(-6) g/ml. The refractory period of the cardiac muscles was prolonged in the presence of alprenolol (10(-7) to 10(-6) g/ml) or carteolol (10(-6) to 10(-5) g/ml). The refractory period of the papillary muscle was increased by 10(-7) to 10(-6) g/ml of propranolol. The maximum driven frequency (MDF) of the atrial muscle was reduced by 5X10(-8) to 10(-7) g/ml of carteolol or by 5X10(-8) g/ml of alprenolol, and MDF of the papillary muscle by 5X10(-7) g/ml of propranolol. In extremely high concentrations, these compounds significantly reduced MDF of the cardiac muscle. These results suggest that the antiarrhythmic action of alprenolol and carteolol is more predominant in atria than in ventricles, whereas the reverse is true for propranolol.


[Important factors determining human distribution and elimination of diazepam]. The pharmacokinetics of diazepam and its biologically active major metabolite desmethyldiazepam were investigated under clinically relevant situations. Both drugs were measured in the different specimens (blood, plasma, urine, bile) by a specific and sensitive gas-liquid chromatographic assay. The pharmacokinetic data were analyzed by the digital computer program SAAM-25 according to the two compartment open model. In healthy subjects the elimination half-life (T1/2(beta))is dependent on the age of the individuals, which is caused by changes in the apparent volume of distribution. The strong plasma protein binding of 95 to 98 percent determines the low hepatic clearance of diazepam. Only neglible amounts of a dose are excreted unchanged with the bile and into the urine. After multiple dosing with diazepam its T1/2(beta) is prolonged, which is caused by a lowered clearance. Thereby also desmethyldiazepam accumulates, since it is eliminated about three times slower than its parent compound. In patients with dysfunction of the liver (cirrhosis, hepatitis) diazepam and desmethyldiazepam exhibit a prolonged T1/2(beta) and a reduced clearance and the lowered plasma protein binding causes a larger volume of distribution. Animal and in vitro experiments demonstrate, besides species dependent differences, that the elimination of diazepam can be impaired by the accumulating desmethyldiazepam.


[Progress in flexor-tendon surgery].

[Progress in flexor-tendon surgery]. The treatment of fresh injuries with the modified technique of primary suture followed by a dressing in the relaxed position of the suture together with the parallel progress in transplantation have brought a new stimulus to flexor tendon surgery. The possibilities of selecting the best approach in every case have become greater and have been extended to include the field of micro-surgery. Also, technical progress has become more cautious. Each intensive specialization, which must necessarily be limited in its application to only a few centres, obliges all surgeons however, to master and apply the theoretical principles of these advances.


[Natural science and historicity in modern times]. The history of science in modern times is the history of the loss of and gain in historicity. In contrast to the increasing historicizing of nature (as the objective dimension), the knowledge of nature (as the subjective dimension) after a time of parallelism between historical studies and scientific research (Enlightenment) and a time of integration (Idealism/Romanticism) was fundamentally dehistoricized during the positivistic 19th century. The history of science and science itself have since that time fallen apart--consciousness of science is restricted to consciousness of the presence.


[Experiences with a new percutaneously effective antirheumatic preparation]. The new substance Etofenamat (Rheumon Gel) is effective percutaneously in the treatment of rheumatic diseases. The gel is shown to be an effective antiphlogistic and analgetic, especially in vertebragenic diseases. The results of treatment of 81 patients are summarized. In a total of 56 patients, use of Etofenamat led to an evident improvement, for example, freedom from pain. Treatment for up to 9 days, eliminated pain in 73 of the 81 patients. In 28 patients out of 38, the mobility of joints was favourably effected. No side effects were observed.


[Preventive gastrectomies before kidney transplantations]. Gastroduodenal bleeding or perforation occurs in 3.5 to 60% of renal transplant patients. Mortality ranges from 35 to 75%. Experience with prophylactic gastric surgery is based on 61 patients with chronic hemodialysis, who were prepared for renal transplantation since 1974. History, presence of ulcer disease or hyperchlorhydria were assumed to represent a gastroduodenal risk. All 20 patients with such a risk-factor received prophylactic gastric surgery (high risk: selective gastric vagotomy + hemigastrectomy in 12, low risk: selective proximal vagotomy in 6; truncal vagotomy + drainage in 2 cases). Postoperatively there were no serious complications, no lethality.


Isolation of Bacteroides corrodens and Eikenella corrodens from human clinical specimens. Comparative study of incidence and methods of identification. In the course of a systematic search for Bacteroides corrodents and Eikenella corrodens in clinical specimens submitted for microbiological analysis, 61% of the specimens from anal abscesses, 6% of the vaginal specimens and none of the pharyngeal specimens yielded B. corrodens, whereas E. corrodens was recovered from only 9% of the pharyngeal specimens. Some characteristics were found to be useful in differentiating between the two species: B. corrodens strains were strictly anaerobic, cytochrome-oxidase-negative, urease-positive and gelatinase-positive; they were sensitive to lincomycin but resistant to vancomycin. E. corrodens strains on the other hand were facultatively anaerobic, oxidase-positive, urease-negative and gelatinase-negative; they were resistant to lincomycin but sensitive to vancomycin. The pathogenicity of the two species was difficult to assess as in most cases they were recovered from mixed cultures.


[Azlocillin and mezlocillin: two new semisynthetic acylureido-penicillins (author's transl)]. The pharmacokinetic parameters of two new ureido-penicillins (azlocillin and mezlocillin) were determined in 12 healthy subjects after a half-hour continuous infusion of 5,000 mg. The agar diffusion test (test strain Bacillus subtilis ATCC 6633) was used for the microbiological assays. The mean azlocillin serum concentration after the half-hour infusion was 431.0 +/- 75.0 microgram/ml; after eight hours it had fallen to a mean value of 4.7 +/- 2.6 microngram/ml. The mean elimination half-life was 77.5 +/- 10.4 minutes, and the relative distribution volume was 19.4 +/- 1.9% of the bodyweight. At the end of the infusion, mezlocillin showed a mean serum concentration of 426.0 +/- 61.0 microgram/ml and after eight hours an average of 1.1 +/-0.9 microgram/ml; the half-life was shorter (56.9 +/- 9.9 minutes) and the distribution volume lower (14.8 +/- 3.1%) than that of azlocillin. The renal clearance values measured in three subjects during a four-hour continuous infusion were: azlocillin 111.6 ml/min/1.73 m2, mezlocillin 121.5 ml/min/1.73 m2. The kinetic behaviour of the two ureido-penicillins was essentially very similar to that of ampicillin and carbenicillin, 38 patients with bronchopneumonia, cholangitis or urinary tract infections, which in some instances were severe, were treated for an average of 10 days with an average daily dosage of 3X4.0 g azlocillin or 3X5.0 g mezlocillin. 30 patients showed clinical improvement, and in 17 of these the pathogen was eliminated. These therapeutic results appear more favourable than those obtained with the newer aminoglycoside antibiotics (amikacin, sisomicin); in particular the drug was well tolerated.


A review of the medical considerations of the use of tylosin and other macrolide antibiotics as additives in animal feeds. The use of antibiotics in animal feeds is reviewed with special reference to possible problems which might be encountered in human medicine. Emphasis is placed on the macrolide antibiotics, especially tylosin. Even though there can be cross-resistance between tylosin and erythromycin, the increasing use of tylosin in animal feeds has not caused an increase of resistance to erythromycin in human isolates of Staphylococcus aureus. There is no significant evidence that tylosin resistant staphylococci of animal origin have endangered human health. Tylosin is not effective against the gram-negative intestinal flora, which thus does not lead to selection of R-factors by conjugation. From the human medical stand point, there is no reason that precludes the use of tylosin as a fee additive in animal feeds.


Mechanism of Corynebacterium Parvum anti-tumour activity. II. Protective effect in T-cell-deprived mice. The natural resistance to syngeneic mammary carcinoma was reduced by thymectomy alone in adult C3H mice or in mice thymectomized, lethally irradiated and restored with bone-marrow cells (Tx RB). The protective effect of C. parvum was not modified by adult thymectomy but it was reduced in Tx RB mice. The injection of thymic cells to Tx RB mice or the elimination of T cell precursors in bone-marrow cells injected in Tx RB mice did not increase the protective effect of C. parvum. These results were compatible with the hypothesis that T cells do not intervene in the mechanism of C. parvum protective effect.


Plasma levels of di-no-propylacetate and clonazepam in epileptic patients. Plasma levels of DPA and CNP and associated antiepileptic drugs were measured in groups of respectively 106 and 30 epileptic patients aged from 3 to 49 years. A poor correlation between daily oral dose and plasma levels of both drug was observed when the whole group of patients was considered. A better correlation was seen in a group of adult patients who received DPA and PB. Children below 10 years of age disposed both drugs faster than adults and the difference was significant (p less than 0.01) in groups receiving PB as associated drug. Patients medicated with PB and DPA or CNP showed lower plasma levels of both drugs; on the other hand DPA appeared to cause a decrease of PB clearance which was more marked in children. The clinical significance of the fluctuations of daily plasma levels of both drugs are discussed in relationship to the schedule of administration and plasma sampling. Observations on the therapeutic ranges and adverse effect of the drugs are reported.


[How blind is a double-blind trial really? Validity of controlled investigations]. During a double blind trial lasting 4 weeks (Diazepam versus Loxapin), two investigators tried to identify the given drug prematurely. Only the more experienced investigator was able to identify the two drugs (p = 0.01). His judgement was based neither on observation of side-effects nor on a clinical improvement but apparently on a summary of particular subtle effects, the cognition of which requires a specific experience. The use of an adequate comparative drug instead of a placebo in a double blind study does not completely prevent a premature decoding but should reduce this risk and therefore increase the validity of the method.


Tandamine--a new norepinephrine reuptake inhibitor. Clinical, psychometric and quantitative EEG studies in depressed patients. Tandamine (AY-23,946)- a new norepinephrine reuptake inhibitor with practically no serotonin potentiation, MAO inhibition and anticholinergic activity-was administered in doses from 75 to 200 mg to a group of 20 hospitalized depressed patients. Weekly clinical evaluation including the ECDEU global score. Hamilton score and Zung self-rating score demonstrated a slight improvement which became statistically significant between the 1st and 2nd weeks of therapy. While the 'thymoleptic' properties of the drug were weak, a pronouned 'activating/stimulatory' effect of the drug was noted. Psychometric tests showed an increase in attention, concentration and psychomotor activity as well as an improvement in FPI personality dimensions 'depression' and 'inhibition', while the Taylor anxiety score did not reveal any changes. Digital computer period analysis of the EEG demonstrated, 6 h after oral administration of 50 mg tandamine, a decrease of slow waves, a significant increase of fast activity as well as a significant attenuation of the amplitude variability. Such changes are reminiscent of the pharmaco-EEG profile of psychostimulatory drugs and may represent the neurophysiological correlate for the activating/stimulatory properties of tandamine. Our finding suggest that the well-tolerated drug may be of some benefit for retarded depressions.


Typological prediction of response to amitriptyline: a replication study. This paper reports a replication study of prediction of outcome by a cluster-analysis derived four-group typoloy of depression. 143 acutely depressed women were treated with amitriptyline for 4 weeks. In replication of the earlier findings, anxious depressives showed the worst response, hostile depressives and young depressive with personality disorder the best, with psychotic depressive intermediate. The findings provide further validating evidence for the typology, and for the view that nonpsychotic depressives are heterogenous.


Double-blind study with two butyrophenone derivatives: bromperidol vs. haloperidol. The present report deals with a double-blind study comparing the new butyrophenone derivative, bromperidol, with haloperidol as the reference substance. Both substances were found to be highly effective in the treatment of psychotic syndromes belonging predominantly to the schizophrenia group. Certain clues, including the onset of action, seem to be indicative of the superiority of bromperidol. No differences between the two substances were observed with respect to side effects and general tolerability.


[Clear-cell acanthoma].

[Clear-cell acanthoma]. The basis for this review forms a series of 20 new cases of clear cell acanthomas and the literature on this subject. The clinical and histopathological characteristics of this not too rare skin tumor and newer histochemical and electron microscopical findings are discussed. Particular attention will be paid to the question of the position of the clear cell acanthomas within the group of intraepidermal acanthomas and the "clear-celled" tumors of the skin. Presently, the clear cell acanthoma can be classified as a benign epidermal tumor of unknown etiology. It is assumed that the clear cell acanthoma arises through a special disturbance of the differentiation of keratinocytes.


[Clinical picture of erythema neonatorum]. Erythema neonatorum is the most common exanthematous eruption in neonates, which is always associated with an eosinophilic infiltration of the skin. Erythema neonatorum can be present at birth. It is of great importance in the differential diagnosis of pustular eruptions in newborns due to virus or bacteria, of miliaria pustulosa, and of incontinentia pigmenti.


[BCG-ulcer].

[BCG-ulcer]. One of the rare side effects of BCG vaccination and BCG ulcera. Two cases of BCG ulceration are reported.


Yersinia enterocolitica: a review of the bacterium and recommended laboratory methodology. Only in the last few years has Yersinia enterocolitica been recognized as an etiologic agent. The organism, however, is widespread in both the animate and inanimate environments. It has been isolated and recovered from a variety of sources including fecal material, foodstuffs, and water, but vehicles of disease transmission are not fully delineated or understood. The bacteriology of Y. enterocolitica is reviewed and recommended laboratory methodology is described.


Acriflavine violet red bile agar for the isolation of Klebsiella. A medium for the isolation and detection of Klebsiella is described. It contains 0.06% Acriflavine in Violet Red Bile agar (Difco). Klebsiella appeared as 5 to 7 mm mucoid (24 h at 37 degrees C) golden-yellow colonies. Pseudomonas aeruginosa and Enterobacter spp. appeared as small, brown to dark brown colonies. Escherichia coli as well as many other gram negative organisms and gram positive organisms did not grow on this medium.


Comparative study of antistreptolysin O, antideoxyribonuclease B and multienzyme tests in streptococcal infections. The results of antistreptolysin O (ASO), antideoxyribonuclease B (ADB), and a multienzyme test, Streptozyme, with 162 sera from a stratified random sample of Wisconsin school children, and specimens routinely submitted for serodiagnosis of Group A streptococcal infections are described. A review of the results of Streptozyme tests of laboratories participating in the State Division of Health Proficiency Testing Program, is also presented. The results of these various tests have revealed both false positive and false negative results with the Streptozyme procedure. Although there was good correlation of the Streptozyme findings with the ASO and ADB tests results with sera from young children, false positive Streptozyme test results increased with age of the study subjects. Our results with the ADB test on sera from apparently normal Wisconsin children and young adults suggest that the upper limits of normal ADB values, suggested by the manufacturers of commercial ADB test kits, may be too low.


The human gamma-chain variants. A review. The human gamma-chain variants are discussed in relation to their chemical structures and to the importance that their study has had for our present knowledge of the genetics and biosynthesis of hemoglobin chains.


Variation in hemoglobin A2.

Variation in hemoglobin A2. The structure, properties and function of, and some biosynthetic and genetic aspects of, Hb A2 are described. The structural variants of Hb A2 are reviewed and their geographical distribution presented. Hb A2, Hb A2-Flatbush and Hb A2-Babinga are characteristic of negro populations and may have originated in Western or Central Africa. Hb A2-Sphakia is characteristic of Canadian Amerindian and Hb A2-Indonesia of Indonesian/Malay populations. Hb A2-NYU has only been found sporadically and most frequently in persons of Eastern European origin. The other three variants of Hb A2 have only been reported in a single person or in single families. Some conditions which are associated with changes in Hb A2 levels are reviewed.


A streamlined technique for on-grid electron immunocytochemistry. The unlabelled antibody enzyme technique for on-grid immunostaining demands exacting conditions of cleanliness. These can be met with minimal trouble by two technical modifications. Large supplies of all the solutions are millipore-filtered in advance and are then stored frozen in sealed plastic bags and allowed to thaw when required. A disposable, contamination-free surface with depressions to hold drops of staining solution is prepared from a strip of parafilm.


[Immunohistochemical localization of creatinkinase isoenzymes in human tissue (author's transl)]. The use of an immunohistochemical method permits the localization of creatine kinase isoenzymes MM and BB in tissue sections. Frozen sections are first incubated with the specific antiserum and secondly with the soluble antigen under investigation. The antibody fixed creatine kinase can then be visualized by the tetrazolium-salt linked histochemical reaction. In this way CK-BB was found in the smooth muscle and the mucosa of the human colon. In sections of skeletal muscle CK-MM was predominantly localized in the intermyofibrillar space. Membrane bound activity could be demonstrated in the sarcoplasmic reticulum and the surface membrane after elution of the cytoplasmic enzyme. In the human tonsilla CK-BB was localized in lymphatic and epithelial tissues, CK-MM in the muscle fibers. The isoenzyme patterns in single sections of tonsilla were in parallel determined by the immunotitration assay. The results indicate the usefulness of the combined application of histochemistry and immunotitration in serial tissue sections.


Recurrent intestinal intussusceptions in the dog. Intestinal intussusceptions in 2 young dogs recurred within 3 days of initial surgical reduction. Subsequent recurrence was prevented by intestinal resection and creation of multiple intestinal adhesions in 1 dog and by creation of adhesions alone in the other.


Intramammary infections in a dairy herd with a low incidence of Streptococcus agalactiae and Staphylococcus aureus infections. In a dairy herd with a low incidence of intrammary infections due to Streptococcus agalactiae and Staphylococcus aureus, clinical mastitis remained a serious problem despite good control of nonclinical mastitis through postmilking teat disinfection and antibiotic therapy of known infected quarters at the end of lactation. During the 2-year study, the incidence of clinical mastitis was 0.88 cases/cow-year; 32.2% were caused by streptococcal species other than Str agalactiae and 33.5% by gram-negative organisms. Among all new infections detected, 54.1% were caused by streptococcal species other than Str agalactiae and 25.7% by gram-negative bacteria. Among new infections, 41.6% occurred during the nonlactating period or within a few days of calving. Incidence of clinical mastitis was highest in the 1st month of lactation. Among 84 gram-negative infections, 42.8% were caused by Klebsiella pneumoniae, 20.2% by Escherichia coli, and 23.8% by Enterobacter spp. Among the many serotypes of K pneumoniae and E coli, none was predominant.


Resistance of Escherichia coli to penicillins: fine-structure mapping and dominance of chromosomal beta-lactamase mutations. Seven Escherichia coli K-12 mutants with a lowered chromosomal beta-lactamase activity were analyzed genetically. The beta-lactamase-negative mutants isolated from ampA1-carrying strains (resistant to 10 microgram of ampicillin per ml) all carried genetic lesions very close to the ampA1 mutation, which was still present. In an earlier report, two of the mutations mediating a beta-lactamase-negative phenotype (L. G. Burman, T. Park, E. B. Linström, and H. G. Boman, J. Bacteriol. 116:123-130, 1973) were shown to have occurred in the structural gene for beta-lactamase, designated ampC. It is suggested that all beta-lactamase-negative mutants studied here were altered in ampC. The relative order of ampC mutations was (ampC1, ampC8)-ampC9-(ampC12, ampC14)-ampC11, and the gene order was found to be ampC-1mpA-purA. The ampA1 allele was dominant over its wild-type allele but acted only cis and not trans, suggesting that ampA is the promoter or operator region for ampC. A gene dosage effect was found for strains homozygous for ampA+ ampC+ or ampA1 ampC+. Heterozygotes carrying the ampC8 allele on the chromosome showed an apparent derepression of the episomal ampC allele, suggesting a role for beta-lactamase in its own regulation.


Identification of a temperature-sensitive asparaginyl-transfer ribonucleic acid synthetase mutant of Escherichia coli. A temperature-sensitive mutant of Escherichia coli K-12 isolated previously (H. Ohsawa and B. Maruo, J. Bacteriol. 127:1157-1166, 1976) was found to have an alteration in asparaginyl-transfer ribonucleic acid synthetase. This alteration can account for the temperature-sensitive phenotype of the mutant. No evidence was obtained to support the previous suggestion that ribosomal protein S1 is altered in this mutant. Combined with the previous genetic studies, we conclude that the newly defined genetic locus, asnS, for the asparaginyl-transfer ribonucleic acid synthetase maps near pyrD at 21 min on the E. coli chromosome.


Physiological effects of growth of an Escherichia coli temperature-sensitive dnaZ mutant at nonpermissive temperatures. The physiological effects of incubation at nonpermissive temperatures of Escherichia coli mutants that carry a temperature-sensitive dnaZ allele [dnaZ(Ts)2016] were examined. The temperature at which the dnaZ(Ts) protein becomes inactivated in vivo was investigated by measurements of deoxyribonucleic acid (DNA) synthesis at temperatures intermediate between permissive and nonpermissive. DNA synthesis inhibition was reversible by reducing the temperature of cultures from 42 to 30 degrees C; DNA synthesis resumed immediately after temperature reduction and occurred even in the presence of chloramphenicol. Inasmuch as DNA synthesis could be resumed in the absence of protein synthesis, we concluded that the protein product of the dnaZ allele (Ts)2016 is renaturable. Cell division, also inhibited by 42 degrees C incubation, resumed after temperature reduction, but the length of time required for resumption depended on the duration of the period at 42 degrees C. Replicative synthesis of cellular DNA, examined in vitro in toluene-permeabilized cells, was temperature sensitive. Excision repair of ultraviolet light-induced DNA lesions was partially inhibited in dnaZ(Ts) cells at 42 degrees C. The dnaZ(+) product participated in the synthesis of both Okazaki piece (8-12S) and high-molecular-weight DNA. During incubation of dnaZ(Ts)(lambda) lysogens at 42 degrees C, prophage induction occurred, and progeny phage were produced during subsequent incubation at 30 degrees C. The temperature sensitivity of both DNA synthesis and cell division in the dnaZ(Ts)2016 mutant was suppressed by high concentrations of sucrose, lactose, or NaCl. Incubation at 42 degrees C was neither mutagenic nor antimutagenic for the dnaZ(Ts) mutant.


Xylitol and D-arabitol toxicities due to derepressed fructose, galactitol, and sorbitol phosphotransferases of Escherichia coli. d-Arabitol was observed to be toxic to many laboratory strains of Escherichia coli K-12, and xylitol was found to be toxic to an existing E. coli C mutant strain. Fructose-specific components of the phosphoenolpyruvate:sugar phosphotransferase system are required for xylitol toxicity. Selection for xylitol resistance results in Fru(-) strains blocked in fructose phosphotransferase. Introduction of the ptsF or ptsI mutation into a xylitol-sensitive strain eliminates sensitivity. [(14)C]fructose uptake experiments imply that the mutation to xylitol sensitivity, which is co-transducible with ara and leu, results in derepression of normally inducible fructose phosphotransferase. Wild-type strains also become xylitol sensitive if induced by (and then removed from) fructose. Xylitol toxicity is prevented by fructose in both wild-type and mutant strains. Circumstances causing xylitol, a new food additive, to become toxic to an otherwise insensitive wild-type organism have not been reported previously. The d-arabitol-sensitive laboratory strains are galactitol (dulcitol) utilizers, although most other strains are not. Selection for d-arabitol resistance results in Gat(-) strains blocked in a constitutive galactitol-specific component of the phosphotransferase system. A mutation causing d-arabitol sensitivity occurred many years ago in AB284, the parent of AB311, AB312, AB313, and many other strains. d-Arabitol sensitivity also occurs in sorbitol-constitutive strains and is shown, like the previous two instances of pentitol toxicities, to result from a constitutive phosphotransferase, which is blocked in mutants selected for resistance.