The immunoglobulin class responses to oral poliovaccine in coeliac disease. An indirect immunofluorescent technique has been used to study the incidence and immunoglobulin class distribution of the serum antibody response to type II oral poliovaccine in patients with adult coeliac disease and matched control subjects. Nineteen out of thirty-eight (50%) patients with coeliac disease showed an antibody response compared with only seven out of thirty-eight (18%) control subjects. Poliovirus-specific IgA responses were observed in sixteen out of thirty-eight (42%) coeliac patients compared with seven out of thirty-eight (18%) control subjects, while poliovirus-specific IgG responses were observed in only ten out of thirty-eight (26%) coeliac patients and four out of thirty-eight (11%) control subjects. No poliovirus-specific IgM responses were observed. The pre-immunization poliovirus-specific IgA titres of those subjects who showed an antibody response were significantly higher in the coeliac group than in the control group.


Localization of immunoglobulins in intestinal mucosa and the production of secretory antibodies in response to intraluminal administration of bacterial antigens in the preruminant calf. Immunofluorescent studies of intestinal tissues from young preruminant calves demonstrate the presence of two main populations of immunocytes synthesizing IgA and IgM. These cells had infiltrated the lamina propria of the intestine as early as 4 days of age. There was little evidence of any significant involvement of IgG1 in intestinal immune synthesis of calves at this age although activity was demonstrable in the ileum and colon of one calf. In general there were more IgG2-synthesizing cells than IgG1, but these were few compared with the main populations of IgA and IgM cells. Local antigenic stimulus to the intestinal mucosa of young fistulated calves using extracts of heat-killed Gram-negative bacteria produced antibody in the secretions over a period of approximately 3 weeks. A second administration of a similar antigenic dose produced a similar response indicating the requirement for continuous stimuli to maintain a measurable level of antibody secretion. Gel filtration and antiglobulin assays indicated that the antibacterial activity was predominantly associated with IgA and that IgM also played a significant role. Oral administration of bacterial antigens to colostrum-fed calves from 5 to 8 days of age produced a faecal antibody response, indicating that intestinal secretion could be successfully interrelated with the declining passive antibody to maintain an almost continuous level of intestinal antibody in early life.


PHA stimulation of separated human lymphocyte populations. Lymphocyte preparations from peripheral blood and tonsils were separated into populations enriched with T or B cells by formation of rosettes with SRBC and separation of the rosette-forming and non-rosette-forming populations. T cell-enriched populations were also prepared by nylon column filtration. Using these methods preparations were obtained which comprised 80--95% T or B lymphocytes as determined by E-rosette formation and surface immunoglobulin (Ig) staining. PHA responsiveness, measured by [3H]thymidine incorporation, varied between relatively wide limits and was critically dependent on the degree of separation obtained. Relatively pure B-cell populations (less than 12% T cells) from blood and tonsils gave low PHA responses while preparations from blood still containing 24--38% T cells gave responses equal to or even greater than those of unseparated controls (60--78% T cells). T cell-enriched populations (80--86% T cells) responded to an equal or greater degree than controls but more efficient separation (greater than 90% T cells) resulted in markedly reduced stimulation. There was thus no simple correlation between the degree of phytomitogen-induced transformation and the number of T cells present. It is concluded that the low response of relatively pure T-cell populations may be due to depletion of B cells or non-lymphoid cells (or both) during the separation procedures. These observations have implications for the use of PHA stimulation as a measure of T-cell activity in mixed populations such as those of human peripheral blood leucocytes.


Randomized trial of transfer factor treatment of human warts. Dialysed transfer factor, prepared from the leucocytes of a donor whose warts had undergone recent spontaneous regression, was used in the treatment of a child with the Wiskott--Aldrich syndrome. The child then had a spontaneous regression at multiple warty areas. A similar relationship was seen in four otherwise healthy patients in a pilot study. A randomized double-blind study of thirty patients failed to confirm a causal relationship between the transfer factor therapy (equivalent to 2-1 X 10(8) leucocytes) and wart regressions. The need for randomized trials of transfer factor therapy for diseases with a variable natural history is emphasized.


Leukocyte responses to acute renal transplant rejection. A retrospective study of the response of the leukocyte count to renal transplant rejection was performed in 159 rejection episodes. Results of the study showed that 1. contrary to what is commonly thought leukocytosis is the least common response to acute rejection and a decrease in leukocyte count is far more common, 2. a sudden decrease in leukocyte count in a previously stable patient can be an early sign of rejection, 3. there is a significantly greater incidence of graft loss in rejection episodes characterized by leukopenia and 4. the risk of infection following rejection is greatest in patients with rejection characterized by leukopenia.


Impairment of phagocytic activity of macrophages as studied by the skin window test in patients on regular hemodialysis treatment. In 7 patients with less than 4 months of regular hemodialysis treatment (RDT), in 28 patients with more than 4 months RDT and in 9 normal subjects, the skin window technic of Rebuck and Crowley was used to study the cellular response and the phagocytic activity of macrophages. Phagocytosis of carbon particles is decreased in the patients with less than 4 months dialysis, whereas it is not significantly different from the normals in those being dialyzed for longer periods.


The efficacy of carbamazepine combinations in epilepsy. The efficacy and bioavailability, and tolerance to carbamazepinee when administered with phenobarbital or phenytoin or with both drugs were evaluated in a prospective, double-blind study of patients whose seizures were not completely controlled by currently available antiepileptic drugs in usually therapeutic dosages as determined by serum levels. During each of four 21-day treatment periods, one fourth of the patients received daily doses of: (1) carbamazepine (1,200 mg) and phenytoin (300 mg); (2) carbamazepine (1,200 mg) and phenobarbital (300 mg); (3) phenytoin (300 mg) and phenobarbital (300 mg); or (4) carbamazepine (1,200 mg), with phenytoin (300 mg) and phenobarbital (300 mg). The treatment periods were separated by 2 wk of each patient's usual prestudy medication. Treatment with all three drugs was the most efficacious for seizure control. Serum carbamazepine concentration was significantly decreased when the drug was administered with either phenytoin or phenobarbital or both.


Superior bioavailability of digoxin solution in capsules. The bioavailability of various formulations of digoxin was assessed after single and multiple doses in a series of crossover studies in human volunteers. Digoxin tablets that were 97% dissolved in 1 hr in vitro were not significantly better absorbed than tablets with a dissolution rate of 78%. A solution given in capsule form had greater bioavailability than tablets of 97% dissolution rate; serum and urinary glycoside levels after 0.4 mg doses of the encapsulated solution were similar to those attained after 0.5 mg doses of tablets with dissolution rates of 78% and 97%. The bioavailability of the solution in capsule form exceeded that of equal doses of the same solution given as a liquid or that of a standard elixir. No increase in gastrointestinal or cardiac toxicity was detected. Inter- and intrasubject variation in bioavailability was not decreased. Above a certain level, dissolution rate is no longer the limiting factor in digoxin absorption. The mechanism of the enhanced bioavailability of concentrated liquid digoxin in capsule form remains to be determined. Such a preparation deserves further consideration as a possible replacement for digoxin tablets.


Assay of aspirin and naproxen analgesia. To establish the relative potency of naproxen and aspirin for oral analgesia, a 4-point, noncrossover bioassay with placebo control was undertaken with 197 patients. Subjective-response methods were used to determine two measures of postoperative analgesia over a period of 6 hr. With reasonable confidence for an oral analgesic assay, we found 220 mg of naproxen to be equivalent to 600 mg of aspirin for pain relief and 330 mg of naproxen to be equivalent to 600 mg of aspirin for decreased pain intensity.


The nylon column dye test: a possible screening test of phagocyte function. 1. A simple quantitative test has been developed to investigate phagocyte function. 2. This test is performed by the addition of Nitroblue Tetrazolium to whole blood, followed by the isolation of leucocytes on a column of nylon wool. Dye reduction by phagocytes is apparent as a blue coloration of the column due to the formation of formazan. The formazan can be extracted from the column and measured spectrophotometrically. 3. The formation of formazan was found to be directly related to the number of phagocytes in blood. 4. Two patients with chronic granulomatous disease gave abnormal results, suggesting that the test procedure may be of value as a screening procedure for this disease.


"Mental illness" and "disease": outmoded concepts in alcohol and drug rehabilitation. Are the addict and alcoholic mentally ill? By traditional standards, yes. The validity of the "mental illness" concept and its associated psychiatric labeling process, however, is challenged. A synthesis of the human ecological systems and third-force frames of reference is presented as a viable alternative to the medical "disease" model of alcoholism and drug dependency. According to the proposed disease model, "alcoholism," "addiction," and "mental illness" are considered to be modes of coping with pain and anger associated with a person's participating in social systems that frustrate self-actualization and diminish self-esteem. Combined treatment of "alcoholics," "addicts," and "nonaddicted psychiatric patients" is supported with qualifications.


Time constant of thermistors and its role in thermodilution methods. Thermolution serves for measuring either the cardiac output or the size of cardiac ventricles. In the former instance, an accurate determination of the area under the dilution curve, and in the latter, a true tracing of the thermal changes with time are of special importance. Because the size of the area is independent of the time constant, and a curve devoid of marked stroke waves is better suited for automatic integration, thermistors with time constants of 0.8 to 1.0 sec are chosen for this purpose. A reliable determination of the time constant is described. In measurements of the size of cardiac ventricles, a true course of thermal changes with time can be recorded with a thermistor possessing a very short time constant: 0.01 to 0.02 seconds. It is very difficult to make such thermistorized probes or catheters. It is easier to make thermistors with a time constant of 0.1 o 0.2 sec, well insulated, which may serve both requirements. By compensation, the time constant is reducible to 0.01 to 0.2 sec, making possible a true-shape recording of thermal changes for the measurement of the ventricular size, or the constant can be prolonged to 0.8 to 1.0 sec for instrumental integration. If both the shape and the time course of the thermal changes have to be truly recorded, then a non-insulated thermistor with a very short time constant has to be used.


An historical survey of radiobiology and radiotherapy with fast neutrons. The treatment of cancer using fast neutrons was first attempted from 1938 to 1942, only a few years after the identification of the particle in 1932. The radiobiological information which was available at that time was both inadequate and contradictory, and provided no definite rationale for using neutrons in preference to X-rays. The doses given were often too high, causing many patients to suffer severe late reactions. As a result, further attempts to use fast neutrons in radiotherapy were abandoned for nearly 30 years. Interest in the use of fast neutrons was stimulated again by the elucidation of the oxygen effect and the discovery that it was less for neutrons than for X-rays. Thus tumours containing hypoxic cells would be less protected against neutrons. Also the reduced repair of sublethal damage with neutrons provided at least a partial explanation of the miscalculation of dose in the early trial. This was confirmed by means of a series of experiments on pig skin, from which it was also concluded that late damage was not more severe after neutrons, compared with X-rays for a given degree of early damage. A new clinical trial began in 1966, and the results so far are encouraging. In order to relate radiotherapy experience with X-rays to neutrons, it is necessary to measure the relative biological effectiveness (RBE) of neutrons. This has been done for skin of man, pig, mouse and rat. Because of the smaller recovery from sublethal damage after neutrons, the RBE increases as the dose per fraction decreases, but the relationship between RBE and dose per fraction is the same for all four species. Similar information, but only for rodents, has been obtained for a variety of other normal tissues with both cyclotron-produced and monoenergetic 14 or 15 meV neutrons. Experiments with animal tumours have indicated that there might be a wide variation in RBE from tumour to tumour due both to the presence of hypoxic cells and to differences in their capacities to recover from sublethal damage after X-rays and neutrons. The largest series of experiments on one tumour shows that whereas certain fractionation techniques with X-rays may produce a poor tumour response for a given level of normal tissue damage, all the neutron regimes produced a similar, close to optimum result. There is no evidence from which to expect any special dangers from neutron irradiation, and their likely advantage is that they may provide a more reliable method of radiotherapy as well as sterilizing some tumours which are normally resistant to X-rays.


Colicin typing of Shigella sonnei isolated in Northern Taiwan. Eighty-eight strains of Shigella sonnei isolated in northern Taiwan during the period of 1970-1974, were studied for their colicin types by a modified method of Abbott and Shannon. As a result, 67(76%) typable and 21(24%) non-typable strains were found. The prevalent colicin types of S. sonnei in northern Taiwan were types 15(52%) and 6(15%), followed by types 8(3%), 11(3%) and 12(3%). Other types were not found.


Escherichia coli isolation from raw milk in central and southern Taiwan and their susceptibility to drugs. From September 1972 to May 1973, 285 raw milk samples and 25 samples of high temperature treated retail milk were examined for Escherichia coli. High temperature treated mild was negative but from raw milk samples 320 strains of E. coli were isolated, among which 25 were enteropathogenic. Using Poly A, Poly B and nonovalent antisera whose pathogenic strains were classified into 6 serotypes: 0-26, 0-55, 0-119,0-125, 0-126 ad 0-128. About half (12) of them belonged to 0-119. Three pathogenic E. coli were sensitive to all of the 8 antibiotics tested while others were resistant to 1-3 antibiotics. Among the 295 nonpathogenic strains, 115 were sensitive to all antibiotics whereas the rest were resistant to 1-5 kinds of antibiotics.


A comparison of dextro-amphetamine and racemic-amphetamine in the treatment of the hyperkinetic syndrome or minimal brain dysfunction. In a double-blind trial of placebo, dextro-amphetamine, racemic-amphetamine, and methylphenidate, each used for a week, in 48 children with the diagnosis of Minimal Brain Dysfunction or Hyperkinetic Syndrome, it was found that although on the average dextro-amphetamine as well as methylphenidate was significantly superior to racemic-amphetamine, with side effects about the same, in some cases racemic-amphetamine was superior to both dextro-amphetamine and methylphenidate. In 20 cases, improvement was about the same for both the dextro and racemic forms; of these 20, side effects were absent for both in 10 patients; dextro-amphetamine showed fewer side effects in 3 patients, and racemic-amphetamine showed fewer side effects in 7 patients. In 20 other patients, dextro-amphetamine resulted in greater clinical improvement than racemic-amphetamine, while in 7 cases the reverse was true.


Liquid lithium vs. solid lithium: an open, cross-over, pilot study comparing oral preparations. This paper suggests some of the possible advantages of a liquid lithium preparation and describes an open, cross-over, pilot study comparing solid (capsule or tablet) with liquid lithium salt preparations in terms of obtained blood levels and side-effects for a nonmanic-depressive prison population.


Isoniazid (I.N.H.) in the treatment of depressive syndrome: a pilot trial. An account of a pilot trial of Isoniazid as an anti-depressant is given. About 70% of the total patients improved and above 50% of them improved markedly. The improvement was sustained even after withdrawal of the drug. No patient worsened and the drug was very well tolerated on the whole. These preliminary results of a pilot trial amply justifies further exhaustive study of this drug, particularly in depressions.


Self-training of new eating behavior for weight reduction. The problems of obesity are well documented, but few medical treatment programs have proven successful. Recently developed behavioral techniques have offered promise in the treatment of obesity. However, the time invested by a therapist limited their practical use and adoption by the general physician. A pilot study was conducted which employed a brief period of explanation of behavior modification techniques, development of an individualized program of eating behavior and recording of weight changes to provide feedback on progress. The program is carried out by patients at home with a minimum of physician supervision. The results indicate that such an approach is feasible and successful. Comparing this program to other programs is difficult because of the variability in reporting data and results.


[Long-term follow-up of children with urinary tract infection: bacterial spectrum, E. coli-serotypes 0, relapse, and reinfection (author's transl)]. Within 4 years 1400 children were investigated for urinary-tract infection in a long-term study. Children with manifest infection were treated and followed-up. In 59 children with chronic pyelonephritis 159 recurrences were observed: 146 were reinfections (change of organism) and 13 relapses (organism unchanged). Serotyping of 0-antigens showed differences between children with chronic pyelonephritis and children with a single exacerbation within the observation period. Reinfection with resistant bacteria mainly occurred shortly after cessation of therapy.


[Diagnostic approaches in systemic lupus erythematosus (author's transl)]. 333 sera from 295 patients were tested for antinuclear antibodies (ANA) by indirect immunofluorescence, and for their binding capacities towards "native", double stranded DNA (anti-nDNA) by a commercially available radioassay kit. 63 out of 66 SLE sera were ANA positive, and 42 were anti-nDNA positive. 267 "non-SLE" sera were also tested, originating from patients with chronic aggressive hepatitis (77), rheumatoid arthritis (86), scleroderma (40), pseudo-LE syndrome (35), and various other "collagenous" diseases (29). 120 of these 267 sera were ANA positive, while only 16 (6%) gave elevated anti-nDNA values. Thus it appears that this anti-nDNA test kit is a helpful tool for the serological diagnosis of SLE.


Simple analgesics.

Simple analgesics. A number of drugs are available that act fairly specifically as "mild" analgesics, although this description by no means implies that their clinical effectiveness is limited to the relief of slight pain and trivial disability. They are effective by mouth and their action is mediated peripherally. Among the possible mechanisms of action, the inhibition of prostaglandin synthesis is currently regarded as most likely to be relevant. Some centrally acting drugs  of the narcotic analgesic type, such as codeine and dextropropoxyphene are effective orally; they are usable in the same way as other mild analgesics and may be preferable for some types of pain. Many problems arise in the assessment and comparison of mild analgesics, both experimentally and clinically. Subjective assessments may be made on a pain scale by the patient himself, or by a trained observer. Individual variations are all-important, and the limitations of controlled trials need to be remembered. Alternative drugs and mixtures have little advantage over aspirin, but specific drug tolerance, in the long term, varies from patient to patient. Gastric irritation is most likely to occur with aspirin in the presence of chronic dyspepsia or acute precipitating causes such as alchoholic gastritis. Allergy also occurs in some susceptible individuals. The risk of renal damage with phenacetin is increasingly appreciated, and the possibility of hepatic damage from paracetamol is now recognised. Other side-effects and interactions are summarized in the review, and some notes are given on therapeutic and non-therapeutic use.


[Allergic reactions of the mouth mucosa in patients wearing dental prostheses]. Due to the increase in the frequency of allergic disorders it is necessary also to consider allergic reactions in the case of unspecific lesions in the oral region. Intolerance of drugs, resins, and metals can be demonstrated by elimination and exposure tests, but also by epicutaneous tests. Diagnostic measures and therapy vary with the type of allergy in each case.


[The patient's intolerance of dentures from the internal medicine veiwpoint]. Similar complaints these found in cases of denture incompatibility (itching of palate and tongue, dry mouth, sores in the mouth as well as other unpleasant sensations) may be observed in many disorders of internal medicine. Therefore considerable difficulties are often involved in the differential diagnostic considerations and delimination of such complaints.


Prolactin and growth hormone secretion in chemically induced and genetically obese mice. Experiments were performed to determine PRL and GH concentrations in mice rendered obese by chemical means and to compare these concentrations with those of mice obese as a result of genetic mutation. Basal levels of serum PRL and GH were generally lower in gold thioglucose (GTG) and bipiperidyl mustard (BPM)-treated obese mice compared with lean controls. In the pituitary gland, the hormonal changes varied from lower or unchanged levels of PRL and GH shortly after drug injection to very high concentrations of PRL (but not of GH) a year later. However, when the mice were challenged with perphenazine, a drug that causes prompt release of PRL, GTG and BPM-obese mice released 2-5 times as much PRL as did lean controls, suggesting an impairment in the hypothalamic control of PRL secretion in GTG/BPM-obese mice. Basal levels of PRL and GH in genetically obese (ob/ob) mice of both sexes were also lower than those in their lean relatives (?/+). This was true for both serum and pituitary concentrations of the two hormones, the only exception being pituitary GH concentrations in females which were higher than or equal to those of controls. However, unlike GTG and BPM-obese mice, genetically obese mice released very little PRL compared with their lean relatives when stimulated with perphenazine, which suggested an insufficiency of pituitary function in ob/ob mice. The results demonstrate abnormalities in the secretion of PRL and GH in obese mice of both types.


The stimulatory and inhibitory function of theophylline on amino acid-induced insulin release: studies with the perfused rat pancreas. The effects of 5 mM theophylline on L-phenylalanine, L-alanine, and L-leucine-induced insulin secretion were studied using the isolated perfused rat pancreas, varying the sequence of the stimuli. Theophylline in the absence or presence of 3 mM D-glucose caused a small amount of insulin release with a slow onset and a slight and steady increase. Theophylline had no effect on phenylalanine (5, 10, or 20 mM)-induced insulin release independent of the sequence of stimuli. Alanine (20 mM), in the absence of glucose, had no significant insulin stimulatory action. When theophylline was added during the alanine perfusion only a small insulin release, comparable to that given by theophylline alone, could be observed. However, superimposing alanine on a theophylline perfusion led to a potentiation of insulin release. The leucine-induced insulin secretion was significantly altered by the addition of theophylline. At a low concentration of leucine (5 mM) theophylline caused potentiation of leucine-induced insulin secretion. At 10 and 20 mM leucine, theophylline led to a rapid concentration-dependent inhibitory period, followed by a potentiation in the case of 10 mM leucine, and by a restoration of the secretion rate at 20 mM leucine which did not exceed the secretion rate of 20 mM leucine alone. Subsequent removal of theophylline caused a marked "off effect." When 5 or 20 mM leucine was superimposed on a theophylline perfusion, a marked dose-dependent potentiation of the biphasic leucine-induced insulin release and no inhibitory phase could be observed. From these data it must be concluded that the effect of theophylline on the insulin secretory reposure of the beta cell to theophylline and the stimulus. Possible explanations for these phenomena are discussed.


Ewe luteal function influenced by pulsatile administration of synthetic LHRH/FSHRH. The influence of repetitive administration of synthetic LHRH/FSHRH or saline (S) on 17beta-estradiol (E2)-induced precocious luteal regression in the ewe was examined. Ewes were pre-treated on days 10 and 11 of the estrous cycle with either 750 mug E2 (total dose = 1.5 mg) in oil or with oil (O) alone. Treatment involved in delivery of 10 mug of synthetic LHRH/FSHRH in saline or an equal volume of saline only, administered at 2-h intervals beginning on day 12 of the estrous cycle and continuing through the succeeding 72 hours. During the period of LHRH administration, the serum LH patterns in the O-LHRH and E2-LHRH groups were characterized by rhythmic fluctuation, rising in response to LHRH and falling prior to the subsequent treatment injection. Throughout the course of the treatment period, the serum LH levels in the O-LHRH group were consistently higher than those in the E2-LHRH group. No increase in serum LH concentration was observed in the saline-treated animals. The mean luteal weight and mean luteal progesterone content at the end of the 72-h period were not significantly different between the O-S and E2-LHRH groups (543 +/- 88 vs. 455 +/- 126 mg and 13.1 +/- 6.2 vs 16.0 +/- 9.7 mug, respectively). Both luteal weight and progesterone content were increased (P less than .01) in the O-LHRH group (1089 +/- 87 mg and 47.5 +/- 3.1 mug) and significantly reduced (P less than .05) in the E2-S group (309 +/- 49 mg and 5.2 +/- 0.1 mug) compared with those of either the O-S group or the E2-LHRH group. Thus LHRH treatment increased mean luteal weight and mean luteal progesterone content while E2 pre-treatment depressed the same parameters. These data suggest that pulsatile administration of synthetic LHRH is able to elevate serum LH levels to an extent sufficient to counteract both natural luteolysis and premature luteal regression induced by E2 treatment.


Somatostatin inhibition of glucose-, tolbutamide-, theophylline, cytochalasin B-, and calcium-stimulated insulin release in monolayer cultures of rat endocrine pancreas. Somatostatin inhibited insulin secretion stimulated by glucose, tolbutamide, glucose-theophylline, glucose-cytochalasin B, and calcium in monolayer cell cultures of neonatal rat endocrine pancreas. Both 2-deoxyglucose-inhibited glucose-induced insulin release and basal insulin secretion occurring at glucose 1.7 mM were further reduced by somatostatin. In the presence of somatostatin, 1.0 mug/ml, insulin secretion due to glucose, tolbutamide, or glucose-cytochalasin B were inhibited to levels below the basal secretion seen with glucose 1.7 mM. However, insulin secretion stimulated by calcium, and especially by glucose plus theophylline, remained considerably above basal insulin levels, even with somatostatin 1.0 mug/ml. For all stimuli except calcium, at lower concentrations of somatostatin (0.001-0.10 mug/ml) but not at somatostatin 1.0 mug/ml, increased stimulus concentration partially reversed inhibition by somatostatin. For calcium, even at somatostatin 1.0 mug/ml, insulin release was greater when the calcium concentration was raised. Since net calcium uptake by the beta cell or intracellular translocation of calcium within the beta cell from an organelle-bound pool to a cytoplasmic pool may trigger insulin secretion through interaction of calcium with the microtubular-microfilamentous system, we suggest that the inhibition by somatostatin of calcium influx would explain our findings.


Differential effects of hypothalamic deafferentation upon luteinizing hormone-releasing hormone in the median eminence and organum vasculosum of the lamina terminalis. Complete deafferentation of the medial basal hypothalamus (MBH) was performed in male rats to ascertain the location of the cell bodies of nerve terminals containing luteinizing hormone-releasing hormone (LH-RH) in the external layer of the median eminence and the organum vasculosum of the lamina terminalis (OVLT). Radioimmunoassayable LH-RH in the MBH decreased to one-fourth of the control value 10 days after complete deafferentation, whereas, no change was observed in an area coextensive with the OVLT. Immunohistochemical staining of LH-RH appeared less intense within the median eminence of deafferented animals but was normal in the OVLT. These results suggest that the cell bodies of neurons producing LH-RH found in terminals in the OVLT lie outside the MBH. The possible explanations for the decreased LH-RH content in the median eminence are discussed.


Partitioning of trace metals in selective chemical fractions of nearshore sediments. A series of chemical extraction procedures are used to obtain data on the partitioning of trace metals among the various geochemical phases of sediments. These components include intersititial water, solubility of solid minerals, ions on exchange sites, metal carbonates, easily reducible phases, organics and sulfides, iron oxides, and lithogenous (mineral residual) fractions. In general, a mass balance of less than 10% deviation can be obtained. Experimental results show very small fractions of trace metals to be in the form of interstital water or soluble ions. Trace metals in the exchangeable phase are almost negligible, and those in the mineral residual phases range from 2.5% Cd for one sediment to 98% Cu for another. The non-residual trace metals content is found to increase with decreasing sand content.


Solvent extraction of organomercury compounds with quaternary amines. Methylmercuric chloride and phenylmercuric acetate extract very efficiently with quaternary amines dissolved in diethylbenzene or other simple solvents. The extraction is effective from either alkaline or acidic solution. The technique considerably extends the scope of conventional solvent extraction practice to permit the extraction and concentration of total mercury in environmental and biological matrices.


Actions and interactions of dipropylacetate and penicillin on evoked potentials of excised prepiriform cortex of guinea pig. Slices from guinea pig brain containing the lateral olfactory tract (LOT) and the prepiriform cortex were studied in vitro. Field potentials, evoked by stimulation of the LOT, were recorded extracellularly. This field potential comprises a compound action potential, a surface negative wave (identified as EPSP), and superimposed positive peaks ("population spikes" or PSs) reflecting postsynaptic activity. In a previous article the penicillin-induced increase of EPSP and of both amplitude and number of PSs was described. Now we are reporting the slight depression of EPSP and PSs and the prevention of the appearance of penicillin-induced PSs by an antiepileptic drug sodium dipropylacetate (Depakine). The effect was dose-dependent. Models explaining the effects of penicillin and dipropylacetate are discussed.


Concentration of dipropylacetate in plasma. Dipropylacetate is absorbed rapidly and attains a maximum concentration in serum 1 to 3 hr after ingestion. Since its half-life is on the order of 8 to 10 hr, it must be prescribed three times a day, every 8 hr. It reaches a stable concentration in blood within 48 hr after treatment is begun (by contrast with diphenylhydantoin and phenobarbital) and might therefore be useful when seizures must be controlled quickly. The serum concentration can be altered by some other anticonvulsant drugs, but these interactions should be studied in more detail. The serum concentration varies considerably in the course of a day and from one day to the next in the same subject, which makes it difficult to adjust the blood level. Depamide is rapidly transformed in vivo to dipropylacetate.


Specificity of ATP-dependent and GTP-dependent protein kinases with respect to ribosomal proteins of Escherichia coli. Two protein kinases differing in substrate specificity were used to phosphorylate the 30-S and the 50-S ribosomal subunits of Escherichia coli. The catalytic subunit from the rabbit skeletal muscle protein kinase phosphorylates proteins S1, S4, S9, S13 and S18 of the 30-S subunit and proteins L2, L4, L5, L16, L18 and L23 of the 50-S subunit with (gamma-32P)ATP as phosphoryl donor. A second protein kinase isolated from rabbit reticulocytes, formerly shown to phosphorylate preferentially acidic proteins and to use GTP as well as ATP, strongly phosphorylated protein S6, an acidic protein of the small ribosomal subunit, and to a lesser extent proteins L7 and L12 or the large subunit. Evidence is presented showing different phosphorylation patterns when either whole subunits or the extracted proteins were used as substrate for the protein kinase. Kinetic studies showed proteins S1 and S4 to become most rapidly phosphorylated. Although most proteins incorporated less than stoichiometric amounts of phosphate, it is shown that with a high excess of ATP L2 bound 1 mol phosphate/mol protein.


The major proteins of the Escherichia coli outer cell envelope membrane. Characterization of proteins II* and III, comparison of all proteins. Protein II*, one of the major Escherichia coli outer cell envelope membrane proteins has been characterized. The protein is heat-modifiable and perhaps due to complete unfolding and/or binding of sodium dodecylsulfate only at higher temperatures the modified protein exhibits a higher apparent molecular weight (33,000) than the non-modified form (28,000). Protein-chemical evidence as well as the behavior of two mutant proteins II* very strongly suggest that this protein consists of a single polypeptide chain and that in the strains studied there is no other major protein with similar characteristics. For another outer membrane protein, protein III (molecular weight 17,000), it has not yet been established if it should be classified as a major protein. Protein III consists of one or perhaps two polypeptide chains. The possibility existed that protein III is bound covalently to lipopolysaccharide, and this has been ruled out. Also, the lipopolysaccharide of the E. coli strains studied does not carry covalently bound protein in amounts anywhere near stoichiometry. N-on-protein substituents were neither found in protein II* nor in protein III. It is concluded that in E. coli B/r and the E. coli K12 strains used there are three major proteins: I, II, and IV; protein III may also belong to this class. There are not more major proteins than these. All four proteins are compared and discussed regarding their unknown functions and their relation to E. coli outer membrane proteins studied by other authors.


Inter-relations between the phospholipids of rat pancreatic islets during glucose stimulation, and their response to medium inositol and tetracaine. 1. Isolated rat pancreatic islets subjected to an increase in glucose concentration from 0.5 to 3 mg/ml showed an increased insulin secretion and 32P-labelling of their phospholipids, especially of phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol, and CDP diglyceride. Neither fructose or inositol produced a similar effect. 2. The enhanced labelling of CDP diglyceride and phosphatidylglycerol was suppressed  by adding inositol (0.1 mg/ml), while the phosphatidylinositol labelling was increased still further. 3. Tetracaine (0.5 mM), an antagonist of insulin secretion, inhi-ited phosphatidylinositol synthesis while phosphatidylglycerol, CDP diglyceride and phosphatidic acid formation were markedly increased. These effects on phospholipid synthesis were substantially reversed by adding inositol to the medium. Tetracaine also prevented the catabolism of phosphatidylinositol in prelabelled islets but this inhibiton was not reversed by inositol. 4. Inositol addition did not affect insulin secretion in glucose-stimulated islets nor secretion in tetracaine-treated islets. This need not exclude a possible role for heightened phosphatidylinositol cleavage in stimulated insulin secretion.


Small-angle x-ray studies on the structure of 16-S ribosomal RNA and of a complex of ribosomal protein S4 and 16-S ribosomal RNA from Escherichia coli. 16-S ribosomal RNA and a complex of ribosomal protein S4 and 16-S rRNA were studied in solution by small-angle X-ray scattering. Concentration series of the 16-S rRNA and the S4 - 16-S-rRNA complex were measured in 37.5 mM Tris-HCl buffer pH 7.4 at 5 degrees C. The following data were determined. The radii of gyration for the 16-S rRNA and S4 - 16-S-rRNA complex were R = 17.6 +/- 0.6 nm, respectively. The two respective values of the radii of gyration of the cross-section were Rq,1 = 8.42 +/- 0.1 nm and 8.33 +/- 0.3 nm, and Rq,2 = 0.988 +/- 0.03 nm and 0.996 +/- 0.03 nm. The largest diameters of the 16-S RNA and S4 - 16-S-RNA complex were L = 61.8 +/- 1 nm and 60.0 +/- 1 nm, respectively. Volumes of V = 1570 +/- 60 nm3 were found for both particles. In the Tris buffer used, no significant differences were found between the scattering curves of 16-S rRNA and the complex is a flat elliptical cylinder with the following dimensions: large axis 61.7 nm, small axis 35.4 nm and height 2 nm. The theoretical scattering curve fits the experimental one as long as the shape of the measured curve is due only to the overall shape of the particle. A model equivalent over the whole measured angular range is one built up from a large number of spheres that simulate the known substructure of the RNA. The outer dimensions of this model correspond to those of the flat elliptical cylinder.


Disturbance in the regulation of the type of collagen synthesized in a form of osteogenesis imperfecta. Fibroblasts derived from skin biopsies of a patient with a congenital defect of connective tissue revealed a disturbance in collagen synthesis. The defect was found in the mechanism that controls the type of collagen synthesized. Biochemical data as well as evaluation of immunofluorescent micrographs using collagen type-specific antibodies, suggested that the fibroblasts of this patient synthesized type III collagen, at a time and rate which was not found in fibroblasts from an age-matched healthy individual.


Gram-scale purification of methionyl-tRNA and tyrosyl-tRNA synthetases from Escherichia coli. A procedure is presented for the purification of 3 g of homogeneous methionyl-tRNA synthetase and 1 g of homogeneous tyrosyl-tRNA synthetase from 50 kg batches of Escherichia coli. The procedure permits the isolation of many enzymes simultaneously and the elution positions of seven other aminoacyl-tRNA synthetases, catalase, rhodanese, phosphofructokinase, elongation factor Tu and cytochrome b-562 are indicated. The problems of extraction work on this scale are discussed.


Isolation and properties of an arginine-binding protein from Saccharomyces cerevisiae. Transfer of exponentially growing cells of Saccharomyces cerevisiae epsilon 1278 b to a fresh medium (or simply to distilled water) resulted in the loss of ability to transport arginine (and lysine), accompanied by the release of several proteins from the membrane surface or periplasmic space. Fractionation by ultrafiltration, Sephadex G-50 chromatography and freeze-drying yielded a homogeneous protein (55 mg per 100 g dry weight of cells) with specific binding ability for L-arginine (Kd = 3.8 X 10(-1) M) and L-lysine (Ki = 4.2 X 10(-4) M). The protein contains over 40 amino acid residues and has a molecular weight of about 5,000. In solution, it appears to aggregate as its concentration is raised, thereby decreasing the overall binding capacity for arginine. Addition of the protein to a depleted culture does not restore the transport of arginine. It is apparently the recognition protein for the specific arginine-transporting system of Saccharomyces cerevisiae but it occurs in almost identical amounts in the MG 168 mutant with impaired arginine transport.


Amino-acid incorporation into tRNA fragments and into heterologous combinations of fragments. When the CCA-halves of tRNAAla1(yeast) and tRNAVal1(Escherichia coli) were incubated with the pG-halves of tRNAVal1 (E.coli) and trnaala1 (yeast), respectively, heterologous complexes were detected. When a 10-fold excess of one half was applied, up to 50% of the other half could be complexed. 5--12% alanine and valine incorporation was observed into the heterolgous combinations in which the pG-halves were derived from tRNAAla1 (yeast) and tRNAVal1 (E.coli), respectively. Although the values are small they appear to be significant considering the results of a number of control experiments. The CCA-half of tRNASer1,2(yeast) and another fragment of this tRNA which extends from the dihydrouridine region to the CCA-terminus were inactive in the aminoacylation assay but they could be converted into a form which accepted serine under standard conditions even in the absence of a complementary fragment. One activation procedure involved the addition of MgCl2 to Mg2+-free fragment solutions, the other consisted in a brief heating-cooling cycle of the fragment solutions at low Mg2+ concentrations. With the two procedures up to 20% or up to 40%, respectively, of the maximal serine incorporation were achieved. At 37 degrees C the active conformation of the fragments persisted only for a few minutes. Analogously, the CCA-halves of tRNAPhe (yeast), tRNAAla1 (yeast), and tRNAVal1 (E.coli)could be activated although here the extent of aminoacylation varied greatly from one experiment to the other. Mischarging of the activated CCA-halves of tRNASer1,2 (yeast) and tRNAPhe (yeast) with phenylalanine and serine, respectively, was not observed. The results obtained with the hererologous fragment combinations and with the CCA-halves alone, which at first sight seem to contradict each other, are discussed with respect to the conformational requirements of synthetase-tRNA recognition.


Temperature-sensitive mutants of the yeast fatty-acid-synthetase complex. By genetic complementation analysis, 88 independently isolated temperature-sensitive fatty acid synthetase mutants have been assigned to the six different fas-complementation groups II (fas 1), III (fas 1), Vb (fas 1), VI (fas 2), VIII (fas 2) and IX (fas 2). The complementation groups Va, Vc, Vd, IV and VII observed among nonconditional fas-mutants have not been found among the temperature-sensitive strains studied. From the failure to detect pantetheine-deficient conditional fas-mutants it is concluded that the yease acyl-carrier protein has an exceptionally stable tertiary structure. Furthermore, the lack of temperature-sensitive mutants of complementation group IV possibly indicates that this group specifically represents only nonsense and frameshift mutations. Almost half of the temperature-sensitive fas 1 and fas 2 mutants studied exhigited non-complementing characteristics. These results confirm the existence of non-complementing fas1 and fas2 missense mutations. From this it is concluded that both fatty acid synthetase loci encode multifunctional polypeptide chains rather than several monofunctional component enzymes. The possible existence of an independent acyl-carrier protein, as suggested by the genetic data reported in this study, is discussed. With 10 different temperature-sensitive fas1 and fas2 mutants the dependence of cellular growth rates on growth temperature and fatty acid supplementation was determined. With all mutants studied fatty-acid-independent growth was completely suppressed at non-permissive temperatures (34 -37 degrees C). In fatty-acid-supplemented media, however, these mutants exhibited the same growth characteristics as wild-type yeast cells. In contrast to this, wild-type yeast growth was found to be fatty-acid-independent at all temperatures studied. Other than in vivo, the purified fatty acid synthetase isolated from five different temperature-sensitive fas1 and fas2 mutants exhibited in vitro no increased thermolability compared to the wild-type enzyme. From this it is concluded that the specific conformation of fatty acid synthetase subunits either forms only at the ribosomal level during translation, or that this conformation is stabilized by the assembly of subunits into the multienzyme complex structure.


Amino-acid sequence of lac repressor from Escherichia coli. Isolation, sequence analysis and sequence assembly of tryptic peptides and cyanogen-bromide fragments. The lac repressor from Escherichia coli, composed of four identical subunits with a molecular weight of 37160, was carboxymethylated and fragmented by tryptic digestion and cyanogen bromide treatment. Using ion-exchange chromatography, gel filtration and preparative thin-layer electrophoresis and chromatography 29 of the 30 tryptic peptides were isolated in pure form. Direct Edman degradation and the dansyl-Edman technique were used to determine the sequence of the small tryptic peptides. Special emphasis was put on the sequence determination of the six large tryptic fragments which together account for 177 residues, corresponding to 51% of the repressor subunit with its 347 residues. The large tryptic fragments were analyzed after fragmentation with chymotrypsin, thermolysin and dipeptidyl aminopeptidase I. Thus the sequence of all 30 tryptic peptides could be deduced. The complete sequences of all cyanogen bromide fragments were deduced from peptides obtained by tryptic, chymotryptic and thermolytic digestion of the individual fragments and by automated stepwise Edman degradation of lac repressor and of the large cyanogen bromide fragments. The order of the cyanogen bromide fragments was given by overlapping tryptic peptides. The resulting amino acid composition of the monomer is Asp15, Asn11, Thr18, Ser30, Glu14, Gln27, Pro13, Gly22, Ala44, Cys3, Val33, Met9, Ile17, Leu40, Tyr8, Phe4, Trp2, Lys11, His7, Arg19. The sequence of lac repressor shows no similarities with that of other proteins known to bind to DNA or RNA. The N-terminal 55 residues contain two homologous regions. This part of the sequence which is involved in lac operator binding might have been formed by gene duplication.


Readenylation of polyadenylate-free globin messenger RNA restores its stability in vivo. Using an ATP:RNA adenyltransferase from Escherichia coli, a polyadenylic sequence was resynthesized onto rabbit globin mRNA from which the poly (A) segment had been previously removed. Conditions for obtaining a homogenous reconstituted globin mRNA preparation containing 30 adenylic residues per message molecule were determined. The reconstituted globin mRNA was microinjected into Xenopus laevis oocytes. Its stability was very similar to that of native mRNA.


Heterogeneity of lipopolysaccharides. Analysis of polysaccharide chain lengths by sodium dodecylsulfate-polyacrylamide gel electrophoresis. Lipopolysaccharide preparations from R(rough) Escherichia coli O8-,SR(semirough) Salmonella typhimurium and S (smooth) strains E. coli O8 and Citrobacter 396 were disintegrated with sodium dodecylsulfate and subjected to polyacrylamide gel electrophoresis in the presence of 1% sodium dodecylsulfate. The results obtained were compared with those obtained from the same lipopolysaccharide preparations by degradation analysis. In dodecylsulfate gel electrophoresis the lipopolysaccharide preparation from the E. coli R mutant and the S. typhimurium SR mutant showed one band each (R-and SR-band, respectively) with different electrophoretic mobilities. The lipopolysaccharide preparations from the E. coli O8-strain exhibited two bands, one of which had the same electrophoretic mobility as the R-band and the other was identified as S-band. The lipopolysaccharide preparation from the Citrobacter 396-S-strain exhibited four bands: one R-band, one SR-band and two S-bands. The results showed that wild-type S strains contain more than one type of lipopolysaccharide. They differ in the length of their O-specific polysaccharide chains. The lipid A content of the different lipopolysaccharide was expressed in their electrophoretic mobilities.


Recognition of initiation codons in modified f2 RNA by Escherichia coli ribosomes. f2 phage RNA treated with O-methylhydroxylamine under denaturing conditions loses its ordered structure with consequent exposure of the normally hidden initiation codons. In the presence of Escherichia coli ribosomes and crude initiation factors modified f2 RNA binds about 50 times more f-[3H]Met-tRNA than native f2 RNA. The interaction of native f2[14C]RNA with ribosomes requires initiation factors. The binding of O-methylhydroxylamine-modified f2 [14C]RNA to E. coli 70-S or 20-S ribosomes does not depend on the presence of initiation factors. A significant number of ribosomes deficient in initiation factors interact with a molecule of modified f2 [14C]RNA. Treatment of the resultant polysomal complex with pancreatic RNase yields ribosomes with f2 RNA fragments protected against RNase. Almost all AUG/GUG codons in the f2 RNA are located on the RNase-insensitive ribosome-bound fragments, constituting only 25% of the entire molecule. Addition of crude initiation factors to such ribosomes with fragments of modified f2 RNA promotes binding of f-[3H]Met-tRNA. The resultant complex is fully reactive with puromycin. No binding of Ac-Phe-tRNA takes place under similar conditions.


The major proteins of the Escherichia coli outer cell-envelope membrane. Cyanogen bromide fragments of protein I, composition and order. The cyanogen bromide fragments of protein I, a major protein of the Escherichia coli outer cell envelope membrane, have been isolated and characterized. There appear to be two methionine-serine or methionine-threonine sequences causing incomplete cleavage but complete conversion of methionine to homoserine. Largely due to the existence of these overlapping fragments the order of 5 of the 6 fragments present could be deduced. None of the fragments exhibits any remarkable low degree of polarity, and the tryptic fingerprint of the largest fragment (comprising about 60% of protein I) also does not show any conspicuous large fraction of lipophilic peptides. It is concluded that the domain of protein I that may be buried in the lipid phase of the outer membrane in all likelihood is not very large, and there is, in fact, no definite proof yet that protein I is a membrane protein sensu stricto.


Rapid isolation of highly active RNA polymerase from Escherichia coli and its subunits by matrix-bound heparin. 1. RNA polymerase from Escherichia coli is selectively and strongly retained by a heparin-substituted agarose and can be eluted therefrom by a neutral buffer containing 0.6 M salt. The method is applicable to relatively crude preparations of the enzyme on a preparative scale giving highly purified RNA polymerase in excellent yield. The enzyme obtained by this procedure shows the highest specific activity so far reported and is pure and enriched in factor sigma as indicated by dodecylsulfate gel electrophoresis. 2. Based on the differential affinity of the subunits of the enzyme for the heparin-carrying gel matrix, a method for separation of alpha, beta' + beta and sigma subunits by application of urea and salt-containing buffers is described. Upon recombination and dialysis with urea-free buffer 40-50% of the enzyme activity is restored.


Studies of intracellular thymidine nucleotides. Thymineless death and the recovery after re-addition of thymine in Escherichia coli K 12. In a thymine-deprived culture, the mutant cells (deficient in dTDP-glucose pyrophosphorylase activity and named Ter-15) lose viability at a faster rate, form longer filaments for the first 60 min and lose thymidine nucleotides and dTDP-sugar pools at a faster rate for the first 15 min than those of the parent cells, but the dTDP-sugar pool in the parent cells is maintained at high concentration for the first 90 min during thymine starvation. In the recovery of cell growth after re-addition of thymine into the thymine-deprived culture, parent cells recommence growth immediately, but the mutant cells (Ter-15) show a lag-phase for 45 min after which time their growth recommences. The rate of dTTP synthesis for the first 10 to 15 min after re-addition of thymine to thymine-deprived cultures of parent and mutant (Ter-15) cells is three-fold higher than that of thymine nondeprived culture (control), but the rates of dTMP and dTDP-sugar syntheses are the same as those of the control. The total DNA synthesis after re-addition of thymine is equal to that of the control, and the period of thymine starvation other than the number of viable cells during thymine starvation plays an important role. After separation of the filament cells from normal-sized cells by sucrose gradient centrifugation, the initial rate of DNA synthesis of filament cells is three-fold faster than that of normal-sized cells. These results show that the dependency of DNA synthesis upon dTTP concentration is maintained after re-addition of thymine into thymine-deprived culture.


Evidence for an aminoendopeptidase localized near the cell surface of Escherichia coli. Regulation of synthesis by inorganic phosphate. An enzyme capable of hydrolyzing the substrate L-alanine p-nitroanilide has been found in the various Escherichia coli strains tested. This enzyme has been called aminoendopeptidase since it shows both activities (see accompanying paper). It is released from the cells by osmotic shock and by lysozyme -- EDTA spheroplasting treatment, and 50% of the total activity is directly detectable with suspensions of intact cells. However, the release by osmotic shock or spheroplasting is not as efficient as it is for alkaline phosphatase. This periplasmic aminoendopeptidase is constitutively produced but the differential rate of synthesis is increased 4-fold when the cell growth is limited by Pi. The occurrence of this 'derepression' is simultaneous with that of alkaline phosphatase. Increasing the concentration of inorganic phosphate in the medium has no effect on the constitutive aminoendopeptidase synthesis. The effect of phosphate starvation is specific since starvation for neither nitrogen nor carbon and energy source are effective in derepressing aminoendopeptidase.


Regulation of Escherichia coli K10 aminoendopeptidase synthesis. Effects of mutations involved in the regulation of alkaline phosphatase. The synthesis of alkaline phosphatase in Escherichia coli is controlled by the action of at least four genes denoted phoS, phoT, phoR and phoB. The effect of mutations in the first three of these genes on the synthesis of periplasmic aminoendopeptidase of E. coli K 10 have been investigated. phoR gene product does not seem to be involved either in the constitutive or in the derepressed synthesis of this enzyme. Mutations in phoS or phoT influence the intracellular level of Pi in much the same way as depletion of Pi from the growth medium, and only as a consequence influence the synthesis of aminoendopeptidase and alkaline phosphatase. Point, amber or deletion mutations in the alkaline phosphatase structural gene do not affect aminoendopeptidase synthesis. Thus, alkaline phosphatase and 'derepressed' aminoendopeptidase synthesis are co-regulated by the endogenous level of inorganic phosphate. The way by which this regulation operates is discussed.


The first enzyme of the gal operon in inducible and operator-constitutive strains of Escherichia coli. A comparison of the porperties and amino-terminal sequences of UDP galactose 4-epimerase. 1. UDPgalactose 4-epimerase, the product of the first structural gene of the gal operon in Escherichia coli K-12, has been purified from strain HfrH and from its gal operator-constitutive mutant, HfrH 81-2. 2. The two enzymes are purified by the same procedure, behaving identically throughout. They are identical in sedimentation coefficient, sioelectric point, electrophoretic mobility at pH 8.8 and amino-terminal sequence for the first 30 residues. 3. Slight but reporducible differences were observed between the amino acid compositions and peptide maps of the two proteins. These differences may indicate real differences in the primary sequences of the proteins, but if so they are unlikely to be due to the operator-constitutive mutation because of the identy of the sequences for the first 30 residues. 4. No evidence was found to indicate that the gal operator might overlap with the first structural gene.


Biosynthesis of a specific lipoprotein of the Escherichia coli outer membrane on polyribosomes. With use of polyribosomes isolated from Escherichia coli, biosynthesis of a specific lipoprotein of the E. coli outer membrane was investigated. The products of the cell-free system were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis and immunoprecipitation with anti-lipoportein serum, followed by sodium dodecylsulfate-polyacrylamide gel electrophoresis. It was found that one of the major products was the lipoprotein of the E. coli outer membrane. The lipoprotein was also found to be preferentially synthsized on smaller polyribosomes.


The binding of maltose to 'virgin' maltose-binding protein is biphasic. The biphasic binding properties of the galactose-binding and maltose-binding proteins of Escherichia coli may be important in the functioning of these proteins as recognition components of chemoreceptors. However, Richarme and Kepes [Eur. J. Binding curve of the galactose-binding protein may be the result of isotopic dilution, during equilibrium dialysis, by unlabeled ligand retained by the binding throughout purification. Here the binding of maltose to maltose-binding protein which has never previously been exposed to sugar ('virgin' binding protein) is shown to be biphasic. This implies that the unusual binding properties are attributable to the maltose-binding protein itself.


The tryptophan synthase from Escherichia coli. An improved purification procedure for the alpha-subunit and binding studies with substrate analogues. An improved method is described for the purification of the alpha-subunit of tryptophan synthase from Escherichia coli. The standard manganese chloride and acid-precipitation steps have been replaced by rapid and efficient chromatographic procedures. Indoleethanol phosphate, indoleprapanol phosphate and indolebutanol phosphate have been synthesized. They are not cleaved by tryptophan synthase and are strictly competitive inhibitors versus indoleglycerol phosphate. The inhibition constant decreases as the number of methylene groups in the side chain increases. This may reflect an improved accommodation of the indole and phosphate moienerated by binding indole, indoleglycerol phosphate and indolepropanol phosphate to the alpha-subunit are very similar. This reflects the transfer of the indole moiety to an hydrophobic environment within the active center. The binding of indolepropanol phosphate to the alpha2beta2-complex perturbs the spectrum of pyridoxal 5'-phosphate located in the beta2-subunit. This demonstrates direct or indirect interactions between the component active sites. Bind studies by spectrophotometric titration and equilibrium dialysis with indolepropanol [32P]phosphate show that there is only one binding site per equivalent of alpha-subunit. Complex formation with the beta2-subunit increases the affinity of the alpha-subunit for indolepropanol phosphate, It is a general consequence of protein-protein interaction in this system.


Chemical inactivation of Escherichia coli 30-S ribosomes by iodination. Identification of proteins involved in tRNA binding. 30-S ribosomal subunits are inactivated by iodination for both enzymic fMet-tRNA and non-enzymic Phe-tRNA binding activities. This inactivation is due to modification of the protein moiety of the ribosome. Reconstitutions were performed with 16-S RNA and mixtures of total protein isolated from modified subunits and purified proteins isolated from unmodified subunits. This allowed identification of the individual proteins which restore tRNA binding activity. S3, S14 and S19 were identified as proteins involved in fMet-tRNA binding. S1, S2, S3, S14 and S19 were identified as proteins involved in Phe-tRNA binding. Modified particles shown normal sedimentation constants and complete protein compositions both before and after reconstitution. This suggests that the loss of activity is due to modification of one or more of the actual binding sites located on the 30-S subunit and that restoration of activity is due to structural correction at this site rather than to correction of an assembly defect.


Studies of intracellular thymidine nucleotides. Relationship between the synthesis of deoxyribonucleic acid and the thymidine triphosphate pool in Escherichia coli K12. The two types of mutant strains which show resistance to T-even phage infection have been isolated and been shown to have either a higher or lower ratio of dTDP-sugar to dTTP than that of the parent strains. The one with a higher ratio of dTDP-sugar to dTTP than the parents has a large dTDP-sugar pool and small dTTP pool, and a high level of dTDPG pyrophosphorylase activity. The other one, with a lower ratio of dTDP-sugar to dTTP than the parents, has a small dTDP-sugar pool and large dTTP pool, and a low or deficient level of this enzyme activity. They form an entirely mucoid colony in the synthetic agar plate. Mutant cells (Ter-6 and Ter-21) which have deficient dTDPG pyrophosphorylase activity show 2 -- 3 times higher activity of UDPG pyrophosphoyrlase than that of parent cells. The dTDPG pyrophosphorylase-deficient mutants (Ter-15 and Ter-21) have a 3 -- 4 times higher concentration of dTTP and a faster rate of DNA synthesis and cell division than those of parent strains in growth with external thymine. The dTDPG pyrophosphorylase constitutive mutant (Ter-4) has a 0.5 -- 0.33 smaller dTTP pool and a slower rate of DNA synthesis and cell division than those of parent cells grown in the same medium. In the Ter-15 and Ter-21 mutants, the intracellular dTTP-dependent DNA synthesis rapidly disappeared in thymine suboptimal concentration, but the Ter-4 mutant maintained its dTTP-dependent DNA synthesis over a 20 muM concentration of external thymine. In high concentration (100 muM) of external thymidine, the thymidine effects on the intracellular dTTP concentration do not significantly appear in these enzyme-deficient mutants (Ter-15 and Ter-21). Also, the concentration of intracellular dTTP in the cell growth with external thymidine is 2.5 times greater than that with external thymine in these enzyme-deficient mutants (Ter-15 and Ter-21).


Respiratory failure after liver transplantation. A rapidly growing haemangioendothelial sarcoma of the liver in a twenty-two year old woman was treated by liver transplantation. Disseminated intravascular coagulation resulted in massive blood loss during surgery, and contributed to the death of the patient from respiratory failure on the fourth post-operative day, despite continuous post-operative intermittent positive-pressure ventilation. Other factors leading to her respiratory failure are discussed. There was no evidence of dysfunction in the transplanted liver.


Malaria, favism and glucose-6-phosphate dehydrogenase deficiency. Although glucose-6-phosphate dehydrogenase deficient individuals may suffer (sometimes fatally) from favism, a high incidence of this trait occurs in many Mediterranean populations. This apparent paradox is explained on the basis of a synergistic interaction between favism and G-6-PD deficiency that provides increased protection against malaria compared to that of the G-6-PD deficiency alone. This relationship is analogous to that between various hemoglobins and malaria in that there is selection for a more severe trait if it provides more protection against malaria.


Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope. A rapid method has been developed to visualize cell surface receptors in the SEM. Thus mannan at the surface of Candida utilis cells was localized by stabilized colloidal gold granules coated with either anti-mannan antibodies or Con A.


Suppression of sebaceous gland non-specific esterase activity be electrophilic alpha beta-unsaturated compounds. Sebaceous gland non-specific esterase activity was suppressed following application to mouse skin of 2 electrophilic alpha beta-unsaturated compounds, o-chlorobenzylidenemalononitrile and beta-nitrostyrene, but not by three others.


Visualization of chicken red blood cells in capillaries by immunofluorescence. Chicken red blood cells labelled with specific rabbit antiserum were used for the demonstration of cortical distribution of blood in the dog kidney by means of direct immunofluorescence. Immunization and labelling techniques are described.


Isatin-3-anils as excystment and cysticidal agents against Schizopyrenus russelli. A series of isatin-3-anils (with or without a N-piperidino/morpholinomethyl substituent) have been screened for their cysticidal activity against Schizopyrenus russelli. Their ability to cause excystment has also been studied.


Investigation of the organization of mammalian chromosomes at the DNA sequence level. New developments in DNA sequencing techniques permit rapid progress in the determination of both repetitious and single-copy mammalian sequences. Three distinct families of highly repetitious satellite DNA's from the kangaroo rat Dipodomys ordii have been sequenced. With the MS satellite it was possible to show that the basic repeat sequence and its variants were arranged in a nonrandom order suggesting a hierarchy of repeats. The HS-alpha satellite from D. ordii was shown to resemble the guinea pig alpha satellite, a long term evolutionary persistence inconsistent with previous models. Sequences from hemoglobin mRNA were determined using hemoglobin complementary DNA as template for transcription in vitro. Seven of the largest fragments have been assigned to untranslated regions of the mRNA whereas 15 others have been tentatively located within the structural genes. From correlations with sequences from corresponding regions in the human hemoglobin mRNA's we have been able to make the first direct measurements of the rate of fixation of mutations that do not change the amino acid sequence. The minimum estimate for this rate is greater than the highest previously estimated rates of fixation of neutral mutations (calculated for fibrinopeptide A. A new technique, deoxysubstitution sequencing, which should speed determination of the complete mRNA sequences, is described.


Luteinizing hormone-releasing hormone versus human chorionic gonadotropin: differential effect on the development of ovulatory refractoriness and antibodies. Cyclic intramuscular injections of 25 IU of human chorionic gonadotropin (HCG) at 3-week intervals induced ovulatory refractoriness and HCG antibodies after five to eight treatment cycles. Two of fifty rabbits failed to ovulate following two successive injections of luteinizing hormone-releasing hormone (LH-RH); however, no LH-RH antibodies were detected in the sera of these two animals, suggesting that these observations were due to chance alone. Thus, 0.5 mug of LH-RH injected intramuscularly at 3-week intervals did not induce ovulatory refractoriness or antibody formation after as many as 18 successive treatment cycles.


The induction of ovulation in amenorrheic patients with synthetic luteinizing hormone-releasing hormone: the significance of pituitary responsiveness. The pituitary reserve of gonadotropins was evaluated with intravenous injections of 25 mug of synthetic luteinizing hormone-releasing hormone (LH-RH) in 32 patients with secondary amenorrhea. An obvious individual difference in the magnitude of the serum LH response was noted in these patients. Thus the patients were classified into four groups according to their pituitary responsiveness as expressed by the ratio of the serum LH stimulated peak to the basal level. Patients with values within the range of, or higher than, that of the normal follicular phase were classified as having high response (750% and above); within the range of that of the normal luteal phase as having moderate response (500 to 740%); between that of the normal luteal phase and that of postmenopausal women as having low response (250 to 490%); and within the range of, or lower than, that of postmenopausal women as having no response (240% and below). Among 26 clomiphene-failed patients in this series who were treated again with clomiphene and subsequent injection of LH-RH; 10 of 12 patients (25 or 35 cycles) with high response, 6 of 10 patients (12 of 28 cycles) with moderate response, and 1 of 4 patients (1 of 13 cycles) with low response ovulated. Five patients became pregnant. The results of this study indicate that in amenorrheic women the higher the pituitary response to LH-RH the greater the chance of inducing ovulation. Patients with secondary amenorrhea may thus be classified by assessing their pituitary response to LH-RH, which may be useful in predicting the chance for the successful induction of ovulation.


[Histochemical study of the vasomotor innervation of the internal carotid artery basin]. In the dog, the internal carotid artery and its branches were found to have a well developed nervous plexus with two kinds of nerve fibers: containing either active cholinesterase or catecholamines. In various segments of the basin, both the cholinergic and the adrenergic nerve fibers reveal structural and topographical differences depending on the structural and functional features of the vascular wall. The adrenergic and cholinergic nerve fibers are mainly situated in the adventitia of the internal carotid artery forming a terminal nervous plexus in deep portions, while in the vasa vasorum and in the siphon region they are terminated also in superficial layers of muscular coat. The data obtained suggest that the vasomotor effects of the sympathetic and parasympathetic nervous systems are most obvious in the intracranial part of the internal carotid artery, in the region of convolution, and in the adjacent portions of the artery situated within the cavernous sinus.


[Characteristics of the functional behavior of different cerebral arteries and angiospasm location in them]. For the major arteries of the brain, when their tone is normal, the constrictory responses are much greater than the dilatory ones while the tendency to dilation of the pial arteries is considerably greater than their tendency to constriction; some luminal contraction and no dilation of the intracerebral arteries (in the depth of the cortex) appears even at an increased blood flow. The tendency to a spasm of the cerebral arteries is corroborated by the peculiarities of their functional behavior: the spasm-like constriction of the major arteries of the brain occurs much easier than that of the pial arteries; a spasm of the intracerebral arteries may develop, but only under some local effect.


[Criteria of the optimal functioning of large and small pial artery subsystems]. The geometrical parameters of the pial arterial systems were studied in total microscopic preparations of the pia mater of rabbits. The subsystem of the larger pial arteries (with the diameter over 100 microns) was shown to provide a blood transport with the minimum energy losses. On the other hand, in the subsystem of the smaller arteries (under 100 microns), the effective resistance should be relatively great under normal conditions, but when the blood supply to the brain tissue becomes diminished, the changes of their diameters provide the minimum energy loss. Hence, the main function of the larger pial arteries is the blood transport, while that of the smaller ones -- regulation of the blood supply intensity in respective cortical areas.


[Migration of leukocytes in late syphilis]. Following non-specific inflammation, greater numbers of mononuclears than polymorphonuclear leukocytes were found to migrate into a 'skin-window' in patients with late syphilis. This mononuclear preponderance was statistically significant, the normal control subjects showing a preponderance of polymorphonuclears.


Topical treatment with urea-hydrocortisone in atopic dermatitis. A controlled study against betamethasone 17-valerate. When comparing 1% hydrocortisone in a stabilized 10% urea cream with 0.1% betamethasone 17-valerate cream in a double-blind study on 49 patients with atopic dermatitis or atopic winter feet, betamethasone cream was found to be the most potent. The study also showed that in approximately 60% of the patients the clinical response was equal. It is suggested that the urea-hydrocortisone combination may have its place in the long-term topical treatment of atopic dermatitis on account of its water-binding effect.


Cellular detection of HBsAg by immunofluorescence in liver of HBsAg positive and negative subjects. Demonstration of direct immunofluorescence of HBsAg in the cytoplasm and the nucleus of hepatocytes. The study is based on 77 patients: 32 clinically healthy carriers and 45 patients with acute or chronic hepatitis.


The inexhaustible beta cell.

The inexhaustible beta cell. Repeated intensive pancreatic beta-cell stimulation was carried out in 42 subjects, comprising 22 normal controls, 10 mild to "severe" maturity-onset diabetics, and 10 chronic pancreatitis patients. Each subject received 75 gm. oral glucose twice and 1 mg. glucagon plus 0.5 gm. tolbutamide intravenously three times at short intervals. Each of the three combined stimuli caused almost equivalent marked spikes of insulin release in all experimental groups. The total calculated output of insulin was equivalent to the total daily insulin output in normal subjects. Pancreatitics and those with severe diabetes (fasting blood sugar greater than 120 mg./100 ml.) had qualitatively similar but a quantitatively smaller response. Those with mild diabetes were similar to the normal subjects but had an exaggerated response to the second oral glucose dose, suggesting overactivity of the enteroinsular axis. Despite the inordinate insulin levels, hypoglycemia did not occur.


A nonsuppressible increase of glucagon secretion by isolated islets of high-protein-fed rats. We recently demonstrated increased plasma glucagon but nomal insulin in rats fed a high-protein, carbohydrate-free (HP) diet; however, other investigators have reported that both plasma glucagon and insulin are increased after protein feeding. For this reason, we have investigated the ffects of an HP diet on pancreatic secretion of insulin and glucagon. Male rats were fed an HP or control diet for one, three, or five days, and, at the end of the feeding period, blood was taken for glucose, insulin, and glucagon determinations. Additional animals fed the HP and control diets for up to 10 days were sacrificed, the pancreases removed, and islets of Langerhans isolated. Islets were incubated for 30 minutes in media with glucose concentration of1.7, 8.3, 16.7, or 33.4 mM. Insulin and glucagon secreted into the media were determined by radioimmunoassay. Plasma insulin was markedly reduced after one day of HP feeding but gradually returned to normal by the fifth day. Plasma glucagon was not altered on day 1 but was significantly increased after three days of HP feeding. The I/G molar ratio, which declined precipitously on day 1, increased thereafter but, as shown previously, remained at a level that promotes gluconeogenesis for up to 10 days. Insulin secretion by isolated islets of control and HP rats increased more than 10-fold as medium glucose was raised from 1.7 to 16.7 mM. There was no difference in insulin release by the two groups of islets. Glucagon secretion by HP islest at low medium glucose remained normal during the first five days; however, beginning on day 3 there was gradual loss of the suppressive effect of high medium glucose on glucagon secretion. After one week of HP feeding, glucagon secretion at low medium glucose was doubled and there was complete lack of suppression of the elevated hormone production by high medium glucose. The alterations of alpha-cell function induced by HP feeding are similar to those found in human and experimental diabetes.


Studies on the mechanism of epinephrine-induced hyperglycemia in man. Evidence for participation of pancreatic glucagon secretion. In man, epinephrine induces increases in plasma levels of glucagon, a lipolytic and hyperglycemic hormone. To determine glucagon's contribution to this hyperglycemia and lipolysis, the effects of inhibition of pancreatic alpha-cell responses to epinephrine were investigated with somatostatin and adrenergic receptor blockade. To avoid ambiguities that might result from concomitant changes in endogenous insulin secretion, these studies were performed in juvenile-type, insulin-deficient diabetic subjects. Compared with normal subjects, the diabetics had excessive glucagon responses to epinephrine, which had been infused to attain circulating levels within the range found in man in severe stress. Both somatostatin and propranolol completely prevented glucagon responses and diminished the glycemic response to epinephrine by 40 to 50 per cent. Free fatty acid responses to epinephrine were completely prevented by propranolol but unaffected with somatostatin. Phentolamine had no effect on glucose, free fatty acid, or glucagon responses to epinephrine. These studies demonstrate that epinephrine, via a beta-adrenergic receptor mechanism, causes excessive plasma glucagon elevation in human diabetes mellitus and indicate that this hyperglucagonemia participates in the hyperglycemic, but not the lipolytic, response to epinephrine. Catecholamine-induced hyperglucagonemia may thus provide an additional explantation for the deterioration in carbohydrate tolerance associated with stress.


Stereotaxic neurosurgery in the management of cerebral palsy. Ten children with cerebral palsy are presented on whom stereotaxic operations on the central nervous system were performed with the aim of ameliorating athetosis and spasticity. Tere were seven alert and co-operative children with spastic hemiplegia or diplegia, of whom six received benefit from thalamotomy or dentatotomy. The seventh, a child with diplegia, had improvement of his left lower limb, but the right became worse. One child with spastic diplegia, in whom a thoracic meningocoele had been closed at birth, was not improved by bilateral dentatotomy. Two severely quadriplegic children each had bilateral dentatotomy; one was a child with dystonic and spastic quadriplegia. In both cases the resulting reduction in tone and extensor spasm rendered the nursing of these patients much easier. The place of stereotaxic surgery in the central nervous system in the management of children with cerebral palsy is discussed. We suggest that in selected cases the stereotaxic operation should be performed early in order to gain the greatest benefit. Stereotaxic surgery should be regarded as an integral part of the management which involves close co-operation of paediatrician, physiotherapist, neurosurgeon and orthopaedic surgeon.


The stimulus-secretion coupling of glucose-induced insulin release XIX. The insulinotropic effect of glyceraldehyde. Glyceraldehyde is known to stimulate insulin release. Its influence on various parameters of islet function was investigated in order to assess the possible significance of glycolsis in the insulinotropic action of glucose. In the absence of glucose, glyceraldehyde (5-20 mM), but neither dihydroxyacetone nor glycerol stimulated insulin release in rat isolated islets. The glucose-like effect glyceraldehyde (10 mM) was characterized by a shift to the left of the curve relating insulin release to glucose concentration, without any significant increase in the maximal velocity of the secretory process. In the isolated perfused rat pancreas, glyceraldehyde provoked a biphasic secretory response. Glyceraldehyde-induced insulin release was inhibited in the absence of calcium or in the presence of epinephrine, unaffected by mannoheptulose or 3,3-tetramethyleneglutaric acid, and enhanced by theophylline and cytochalism B. Glyceraldehyde also stimulated to pro-insulin biosynthesis and 45Ca net uptake by isolated islets, the latter effect being apparently due, in part at least, to inhibition of calcium outward transport across the cell membrane. At concentrations of nearly equivalent insulinotropic potency, glucose and glyceraldehyde were metabolized at rates yielding comparable output of both lactate and 14CO2. The data indicate that glyceraldehyde mimics many effects of glucose on islet function, suggesting that the insulinotropic action of glucose may be related to its metabolism through the glycolytic pathway.


Selection of early treatment of myelomeningocele: a retrospective analysis of selection procedures. In an attempt to formulate a procedure for the selection of those children with open myelomeningocele most likely to benefit from early surgery, the records of 163 children have been reviewed. No physical findings, either singly or in combination, were found to distinguish accurately between children with good and poor prognoses, and an alternative selection procedure is proposed which takes into consideration the absence or presence of lacunar skull deformity (a finding strongly associated with mental retardation). Early surgery is not recommended for children with lacunar skull deformity and at least two of Lorber's major adverse neonatal criteria, or children with associated gross congenital anomalies. This selection procedure has been applied retrospectively to the 163 children reviewed and has been shown to predict the (known) outcome of these children much more reliably than existing selection criteria.


Methods of assessing and recording motor skills and movement patterns. The literature in different fields of assessment and recording motor skills and movement patterns in children is reviewed and various methods of movement analysis are discussed from a clinical and instrumental point of view. Their advantages and disadvantages in the study of motor development and in neuropaediatric diagnosis and research are considered. The methods chosen for a special purpose, whether a motoscopic, motometric or motographic technique, depend on what the investigator wants to study and on the available facilities, and is also influenced by the degree of co-operation of the patient. Whatever technique is used, the results obtained must enable comparisons to be made with later assessments as that valid statements about the child's progress become possible.


Physical and biochemical characteristics of the human dysraphic spinal cord. The spinal cords were removed from 29 infants between 17 and 46 weeks gestation, all with various forms of spinal dysraphia. The spinal cords were assessed for weight, length, thickness, size and number of cells, and myelination, and then were compared with 86 control spinal cords. The strongest characteristic of the dysraphic cords was a reduced degree of myelination. Cell size and number generally were appropriate for the weight of the cords. There was a tendency for cord weight, length and thickness to be reduced, though in part this was a reflection of poor over-all bodily growth. In a small group of infants these measurements were increased in comparison with the controls.


Bacterial colonisation and infection in lesions of the central nervous system. A study was made of the pattern of bacterial colonisation during the first month of life in a group of 116 newborn infants with spina bifida. 55 patients subsequently developed a total of 90 infective episodes and in over three-quarters of these episodes there was a correlation between the infecting and colonising organisms. The principal sites of infection were the lesion, the ventricles and the urinary tract, and infection accounted for 27 of the 35 deaths in the series. Some of the factors governing the pattern of colonisation and the development of infection in newborn patients are discussed.


Mycoplasma meningitis in infants with spina bifida. Infants with myelomeningocele are liable to develop bacterial colonisation of the lesion which, in the case of Gram-negative bacteria, often progresses to meningitis because of the infants' immature immune status. The time and origin of the bacterial colonisation usually are uncertain. This study reports seven cases of meningitis caused by Mycoplasma hominis, an organism not previously reported to cause meningitis. The commensal status of Mycoplasma hominis in the vagina of many women, and its isolation from the mother of one patient in this group, suggests that the infant acquired the organism in the birth canal and that infection developed over a period of days. Routine cultures from the sac or wound did not show the presence of organisms until after the child had developed signs and symptoms of meningitis. Commensal organisms from the vagina may lie dormant for several days within the closed myelomeningocele and their more virulent strains may then produce meningitis. It is suggested that neonatal meningitis may be caused by organisms acquired during birth, as well as by those to which the infant is exposed later.


Continence, friends, marriage and children in 51 adults with spina bifida. Fifty-one adults with spina bifida (28 men and 23 women aged between 17 and 56 years) were interviewed in their own homes in South Wales. 17 were completely continent, 10 were incontinent and the remainder were continent to some degree. Seven of the 51 had no close friends but almost half the total had five or more close friends. Of the 29 who were unmarried, nine had a steady relationship but 13 felt that they were disqualified from such a relationship because of their disabilities. 11 men and 11 women were married and a total of 39 pregnancies had come to term, resulting in 32 normal children, five stillbirths (two with anencephaly) and two children with spina bifida. The risk of having an affected offspring when one of the parents has spina bifida (based on the combined London, Munster and Cardiff series) is 1 in 23. It is suggested that the children now growing up who have received 'total care' will be more handicapped than the adults in this series.


The effects of hydrocephalus on intelligence, visual perception and school attainment. A group of 59 children with spina bifida, and a group of matched controls, were given detailed psychological and educational tests at five and seven years. The scores of the control group were similar to the national figures. The scores of the children with spina bifida only (without hydrocephalus) were closest to normal, but still below average. Those with spontaneously arrested hydrocephalus were in the backward range of abilities, and those with shunt-treated hydrocephalus had scores which generally were incompatible with normal levels of intelligence and attainment. Abnormal visuo-perceptual functioning closely correlated with defects in intelligence. The results of the school-attainment tests paralleled the distribution of intelligence but many of the children were found to be functioning below expectation for both age and measured intelligence.


Cerebrospinal fluid pressure-gradients in spina bifida cystica, with special reference to the Arnold-Chiari malformation and aqueductal stenosis. Ten infants with spina bifida cystica were investigated during life by simultaneous ventricular and intraspinal CSF pressure recordings, and 11 post-mortem specimens were studied by means of crystic resin castings of the ventricles. The results indicate marked pressure differentials between the lateral ventricles and intraspinal CSF pathways, which contribute to the moulding of the Arnold-Chiari malformation as a sliding hernia. Similar pressure differentials may exist at the incisura, with moulding of tissues producing blockage of the aqueduct and the subarachnoid pathways. A valvular action was demonstrated, in which baseline pressure differences were exaggerated by the infant's straining (e.g. crying, sucking), and it seems that complete dissociation between the two pressures may develop as a result of periods of partial dissociation. Removal of the meningocele sac increased the peaks in recordings of intraspinal pressure produced by straining. If it is carried out when there is a valvular effect, it seems that it will hasten the onset of established ventriculo-spinal pressure dissociation and uncompensated hydrocephalus by leading to impaction of the Arnold-Chiari malformation in the foramen magnum. The results indicate that even after birth the CSF pathways of babies with spina bifida cystica are in a plastic and changeable condition, with competition between constricting and distending forces. The demonstration that pressures tend to be low in the spine and to become lower before exacerbation of hydrocephalus suggests that all the intracranial manifestations of spina bifida cystica may be due to low intraspinal pressure, which results from initial failure of the neural tube to close during embryogenesis.


A quantitative study of the cell population of the cerebellum in children with myelomeningocele. A quantitative assessment has been carried out on the cell populations and DNA content of the cells in the internal granular layer of the central lobes of the cerebellum from 100 children with myelomeningocele and from 120 normal controls. The method used was a computerised analysis of a television image of the neurones in standardised sections. The cell and DNA complement of the declive and central lobe are only slightly reduced in the cases of myelomeningocele, but there is frequently cellular and DNA loss throughout the pyramid and uvula to the nodule. These findings suggest that the central lobes of the cerebellum are probably genetically normal but have irregular degeneration and arrest of growth, probably as an effect of pressure at the foramen magnum.


The embryogenesis of trypan-blue induced spina bifida aperta and short tail in the rat. The study of litters from 140 Wistar-derived rats injected with trypan blue during gestation leads to conclusion that a myelocele can result from faulty closure of the neural plate and that this is accompanied in many cases by blebs in the paraxial mesoderm. Haematomata usually underlie the open neural plate at an early stage and they form by extravasation of blood from the dorsal aortae into the blebs. Local ventral deflection of the notochord beneath a myelocele probably results from delayed separation of the notochord from the hindgut. Complete failure of separation and abnormality induced in the tail-bud could result in sacral agenesis and/or a short tail. All these malformations may result from varying severity of the action of the trypan blue at different developmental stages.


Lacunar skull deformity related to intelligence in children with myelomeningocele and hydrocephalus. It has been suggested that the presence of lacunar skull deformity in infants with myelomeningocele and hydrocephalus is indicative of later lowered intelligence and therefore is a useful criterion in early selection for treatment. In the present study, retrospective investigations were made of the clinical records and skull X-rays of 169 surviving children who had been admitted immediately after birth with myelomeningocele and hydrocephalus to the Children's Hospital, Sheffield. Adequate X-rays were available for 131 of the children, of whom 97 per cent exhibited some degree of lacunar skull deformity. At five years of age or older, the 131 children were tested on the Wechsler Intelligence Scale for Children. The differences in mean IQ between the various degrees of severity were not large, and it was found that 82 per cent of children with the most severe degree of lacunar skull deformity had IQs above 70. These findings do not support the use of lacunar skull deformity as an important measure in selection for treatment of children with myelomeningocele and hydrocephalus.


Some factors relating to intelligence in treated children with spina bifida cystica. The 83 survivors of a consecutive series of children with spina bifida cystica, born between 1963 and 1971 and treated non-selectively since birth, were assessed by intelligence and developmental testing. In nine of the children who had not required shunts the distribution of intelligence was within the normal range. The need for a shunt was significantly related to the presence of craniolacunae and to the sensory level of the lesion recorded at birth. Seven of the 74 shunt-treated children had suffered ventriculitis, meningitis or septicaemia, and their intelligence was significantly worse than the others. In the 67 shunt-treated children who had not suffered infection, intelligence was significantly related to the thickness of the pallium when the shunt was inserted during the first four weeks of life, and to the sensory level of the lesion recorded at birth. Intelligence was not related to the function of the shunt at time of assessment, to the number of revisions of the shunt, or to the rate of increase in head size during the first four weeks of life. It is concluded that the best indication of later intelligence can be gained at birth from the thickness of the pallium and the sensory level of the lesion.


Study of indirect methods for measuring hepatic blood flow in the isolated perfused dog liver. This study was undertaken to assess the reliability of the constant infusion clearance and extraction technique (CIT) and the single injection clearance method (SIT), to measure hepatic perfusate flow at different levels of liver function. In the 36 isolated perfused dog livers, hepatic perfusate flow(HPF) was monitored by a direct method (DM), and the HPF was simultaneously measured with an indirect method (IM) utilizing bromosulphophthalein BSP) as indicator (20 SIT and 16 SIT). In 16 experiments, absolute ethanol (750 mM) and sodium taurocholate (600 muM) were seperately utilized to impair BSP metabolism. By comparison with DM, in the experiments with an extraction ratio(ER) of BSP lower than 10%, the error of the IM (ml/min) was for the CIT 154.57 +/- 46.55 and for the SIT 107.25 +/- 34.20. This error was significantly greater than in the experiments with an ER greater than 10% (CIT 15.70 +/- 3.88, SIT 8.20 +/- 1.38) p less than 0.01. In the studies with ER less than 10%, no difference was found in the error detected with the SIT (107 +/- 34.20) and CIT measurements (154.37 +/- 46.55); in the experiments with an ER greater than 10%, the error with the CIT (15.70 +/- 3.88) was greater than the one attained with the SIT (8.20 +/- 1.38, p less than 0.10 greater than 0.05). It is concluded that both methods are reliable techniques, for measuring HPF in livers with normal or near normal clearance capacity, but even in these ideal experimental conditions the values for HPF are inacurate when the liver clearance function is moderate or severly impaired.


Ultrastructural changes of the endocrine pancreas in pregnant rats. The ultramicroscopic appearence of the B-cell of the pregnant rat suggests hyperinsulinism of the individual beta cell. In pregnant rats the B-cell contains an increased volume and an increased number of light granules as well as enlarged mitochondria.