Influence of mixtures of taurocholate, fatty acids, and monolein on the toxic effects of deoxycholate in rat jejunum in vivo. The influence of mixtures of taurocholate (TC), oleic acid (OA), caprylic acid (CA), and monolein (MO) on the toxic effects of deoxycholate (DC) in rat jejunum have been investigated using both a closed loop and perfusion technique. DC induced net secretion of water and electrolytes, inhibited glucose transport and transmural potential difference (PD), and inactivated mucosal "total" and (Na+ -K+)-adenosine triphosphatase. Secretion was reversed to absorption when the instilled or perfused solutions were composed of mixtures of DC, TC and OA; substitution of MO or CA for OA produced a similar effect. DC-induced inhibition of PD, glucose absorption, and mucosal adenosine triphosphatase activity was abolished when DC was mixed with TC and OA. Oleic acid emulsions had no effect on secretion induced by DC. Absorption of DC was inhibited from mixed micellar solutions (TC, OA, DC) but not from pure micellar solutions (TC, DC). These results indicate that the presence of taurocholate and fatty acids or monolein within the intestinal lumen markedly modify a number of the toxic effects of DC on jejunal function. The clinical effects of DC on intestinal function in man may therefore depend on the relative concentrations of other bile salts and lipids within the intestinal lumen.


[Report on 700 gynaecologic cases in diagnostic laparoscopy (author's transl)]. The clinical experience in diagnostic laparoscopy is dealt with under the conditions of the Department of Gynaecology and Obstetrics at Erlangen University. Among 700 cases the indications were infertility (34,4%), pelvic pain 27,4%), suspected ectopic pregnancy (7,9%), endocrinologic cases and malformations (18,9%), pelvic mass without symptoms (8,6%) and others (2,8%). The laparoscopic findings are shown. Important diagnostic clues otherwise missed were found in about half of the patients. In 122 women a laparotomy could be disregarded. One should be aware of diagnostic errors especially in ovarian tumors. Biopsy may help in assessment of ovarian function. In tumor diagnosis, however, it is rather dangerous. Difficulties of the methos (2,9%) and complications (1%) are discussed.


Genetic control of major complex histocompatibility antigens. The histocompatibility loci of the MHC can be separated into two functionally distinct types. One, loci first defined by lymphocyte reactivity in MLC,  LD loci, the phenotypic expression of which leads to proliferation of allogeneic T cells, and two, serologically defined, SD loci, products of which act as targets for cytotoxic lymphocytes. Although the SD loci may be definable by both serological techniques and by lymphocyte reactions in CML, and it may well be that the LD loci products will be defined serologically, the functional difference between them is documented by the apparently converse roles in MLC and CML. The LD loci are most effective in leading to MLC stimulation and do not function as targets in CML for reasons discussed elsewhere; the SD loci function poorly it at all in stimulating proliferation in MLC but are excellent targets in CML. A cellular dichotomy may exist in reaction to these different genetic components of the MHC.


[Intrarenal distribution and ATPase inhibiting activity of cuabain in dogs (author's transl)]. Experiments were undertaken to substantiate the hypothesis that the mechanism of the direct effect of ouabain on the renal excretion of electrolytes is the result of inhibition of the transport enzyme, (Na+, K+)-ATPase. In dogs hydrated with saline, an injection of 3H-ouabain into the unilateral renal artery produced a continuing marked increase in excretion of water and sodium from the kidney, but not from the counter kidney. At maximal diuresis -- 90 min after ouabain injection, both kidneys were removed to assay microsomal ATPase activity and determine radioactivity distributed in subcellular structures. It was demonstrated that 3H-ouabain was deposited in the microsome fraction obtained from the injected kidney in concentrations ranging from 10(-7) to 10(-6) M/kg wet weight and (Na+, K+)-ATPase activity of this fraction was inhibited as compared with that of the microsomal fraction obtained from the counter kidney. Since (Na+, K+)-ATPase activity of renal microsomes was significantly inhibited in vitro by more than 10(-7) M of ouabain, ouabain concentration in microsomes obtained from the injected kidney was considered to be sufficient to inhibit ATPase activity. These findings indicate that ouabain diuresis under the present condition is closly related to direct inhibitory effect of ouabain on (Na+, K+)-ATPase activity of microsomes in tubular cells.


Existence in fetal and adult rat of several forms of material reacting with anti-insulin antibody (IRI). The double antibody procedure detects 3 peaks in the elution fractions of adult or fetal rat sera, after passage on Sephadex G50 or G100 columns. Peak A (apparent MW 6000) contains insulin monomer; peak B (apparent MW 10-12000) is tentatively attributed to proinsulin (or proinsulin like substances); peak C (apparent MW 50-100000) is similar to the so called "big big" insulin. During intravenously induced hyperglycemia, the 3 peaks show parallel increases, but, after the disappearance of peaks A and B in streptozotocin treated rats, peak C remains unaltered. Pancreatic extracts and secreta present a very minor and inconstant peak C, the bulk of their immunoreactive material belonging to peaks A and B. A companion paper further discusses the nature of peaks B and C materials.


Possibility of adrenal-testicular interaction as indicated by plasma androgens in response to HCG in men with normal, suppressed and impaired adrenal function. In order to assess possible adrenal-testicular interactions in vivo, adrenal and testicular plasma androgens (testosterone, delta4-androstenedione, dehydroepiandrosterone and dehydroepiandrosterone sulfate) were measured by specific radioimmunoassays before and after stimulation with HCG in men with normal, dexamethasone suppressed and impaired adrenal function. It was found that men with Addison's disease, in whom circulating dehydroepiandrosterone sulfate levels amounted to 1/10 of normal values, had a decreased response of testosterone to HCG. Simultaneously, the Addison patients and the men under dexamethasone had only an increase of delta4-androstenedione but not of dehydroepiandrosterone, while normal men showed an almost equal increase in both precursors under HCG. The results are interpreted as demonstrating that the delta5-pathway in the testis becomes less important under adrenal suppression and in Addison's disease due to a lack of substrate (possibly dehydroepiandrosterone sulfate) from the adrenals.


Human karyotype polymorphism. III. Routine ank fluorescence microscopic investigation of chromosomes in normal adults and mentally retarded children. The results of a routine and fluorescence microscopic investigation of chromosomes are presented in 208 normal adults and 141 mentally retarded children (80 with diagnosis of "mental retardation of unknown etiology" and 61 with Down's syndrome). The occurence of both real chromosome aberrations and chromosome variants are presented, with special attention being paid to the chromosome variants. The data obtained show that in all groups studied there is considerable polymorphism of heterochromatic regions that appears to be dependent on sex and connected with the origin of some pathologic conditions.


Linkage group HL-A-MLC-BF (properdin factor B). The site of the Bf locus at the immunogenetic linkage group on chromosome 6. Genetic linkage between the HL-A and Bf loci could be confirmed in 43 families with 168 offspring.  In 4 families, 5 recombinants out of 82 informative meiotic divisions were observed (r = 6.1%).  The localisation of the Bf marker system was studied in 3 families with crossovers between HL-A and MLC.  From these data the following map order of human chromosome 6 can be proposed: HL-A (1st locus) -- HL-A (2nd locus)--MLC-Bf---PGM(3).  The fact that important components of the classical and alternate pathway of complement activation are governed by genes closely linked with HL-A and MLC loci leads to the proposition to include the Bf system into the Major Histocompatibility Complex in man.


Translocation (13q21q).  Four generation family study with analysis of satellite associations, fluorescent markers, and prenatal diagnosis. A (13q21q) translocation was found in an infant with Down's syndrome.  The 17-year-old mother and the grandmother carried the translocation 45,XX,t(13;21)(p12;q11).  The great grandparents had normal karyotypes.  Fluorescence marker studies suggested that the translocation originated in the great grandmother.  The hypothesis was supported by satellite association studies which showed a significant excess of 13-21 and 13-15 associations in the great grandmother.


X monosomy and 21 trisomy in a sibship.

X monosomy and 21 trisomy in a sibship. The case of a sibship of 4, 2 members of which present aneuploïdy (45,X and 47,XX,21+) is reported.  The paternal grandfather and grandmother are first cousins and there is a large number of centromeric associations in the father.


Sex chromatin and cytogenetic survey of 10417 adult males and 357 children institutionalized in Belgian institutions for mentally retarded patients. A survey of adult male immates of the major institutions for mental patients in Belgium was undertaken in 1965. Out of 10417 males examined 61 had positive sex chromatin (5.8%); 201 had bilateral small testis (1.9%), 4 hypogonad patients carried a D/D translocation. Out of 857 karyotyped children, 2.9% carried an autosomal or sex-chromosomal anomaly other than trisomy G. This prevalence of chromosomal anomalies in children with low I.Q. (less than 50) emphasizes the need for further studies.


On the mutation rate of neurofibromatosis. A genetic study of 124 cases of neurofibromatosis was performed. The contingent of probands was mainly represented by a Russian population, most of the individuals being born in the European part of the RSFSR. Both parents of the probands were examined in only 58 cases, the proportion of sporadic cases in this group being 0.79, as compared to 0.77 for the whole group under study. The existing data evaluated by a direct method are not yet sufficient for a decisive estimation of the penetrance, which, however, cannot be under 80%. Segregation analysis of descendants from particular marriages showed a good correspondance to the hypothesis of Mendelian dominance (32 affected children out of 65). These results analyzed together with those obtained by other authors permit an inference on the full penetrance of neurofibromatosis. The genetic interpretation of sporadic cases as a result of new mutations is presented. The prevalence of neurofibromatosis among the 16-year-old youths was evaluated as 12.8 with 10-(5). This value is suggested to be an estimation of the incidence of the condition in the general population, the mutation rate evaluated by a direct method being equal to 4.4 with 10-(5) divided by 4.9 with 10-minus 5. The increased birth order of probands in sporadic cases (against the theoretical expectation) as well as increased paternal age (as compared with controls) were found to be statistically significant (P equals 0.004 and P equals 0.03, respectively) while the difference in maternal ages was statistically insignificant (P equals 0.008). No statistical relationship between sporadic cases and occupational exposure of parents to deleterious chemical and physical factors was found.


Chromatin structure in Down's syndrome.

Chromatin structure in Down's syndrome. In the chromatin of patients with Down's syndrome, changes are shown to occur in a short-term lymphocyte culture of the human peripheral blood. Some of them are induced by the patient's blood serum and are reversible when this is replaced by normal serum. A 100-fold dilution of the blood serum taken in subjects with Down's syndrome does not produce any changes in the structure of the lymphocyte chromatin of the patients. A similar procedure with the blood serum of healthy donors resulted in a drastic activation of their lymphocyte chromatin. These experiments, and investigations on the effect produced by the blood serum on the model desoxyribonucleoprotein systems, support the suggestion that the changed state of the chromatin in subjects with Down's syndrome is caused by a complex set of components contained in the blood serum, whose degree of dissociation deviates from the normal.


Health services system in Hong Kong: professional stratification in a modernizing society. A characteristic of modernizing societies is the coexistence of modern and traditional professions that claim to perform the same functions for the society. As a result of differential support by the dominant classes and their social values, and by the academic and the political authorities, the modern profession occupies a higher stratificational ranking than the traditional profession. In order to struggle for survival and to compete for more social-political and economic resources, the traditional profession advocates a rational revival, i.e. a rationalization of its social organization and technical contentmin this paper, I have illustrated these generalized statements by analyzing and comparing the profession of modern medicine with that of Chinese medicine in the modernizing society of Hong Kong, These two types of professional services are coexisting in the pluralistic health context of Hong Kong, but the former enjoys greater power, higher prestige and more economic resources. The state of modern medical dominance may be due to its connection with the dominant social value of science and to its support by the "higher" classes and the university and the government in Hong Kong. In recent years; however, there has emerged a revivalistic movement in the realm of Chinese medical care. In significant part, this may be influenced by developments in the People's Republic of China.


Agglutinating serum for distinguishing Staphylococcus aureus of human biotype. Antiserum to Staphylococcus aureus strain 17 was treated with S. aureus strain 61218 until the antibodies against thermostable agglutinogen were removed. The absorbed serum agglutinated phage-typable as well as phageuntypable staphylococci of human biotype, whether recovered from people or from dogs.


A new reliable method for evaluation of foot pad swelling as an experimental in vivo immune phenomenon. Weight determination of replicas of the feet from living mice is described as a new method for evaluating the degree of the experimentally induced foot pad swelling in immunized mice. The new method gives objective, well documentable results and offers advantage to the commonly used assay with the dial calliper gauge when there is need for a good documentation or whenever there is a large number of animals to be tested within a narrow time margin.


An in vitro inhibitor of Na-K-ATPase present in an adenosinetriphosphate preparation. An in vitro inhibitor of Na-K-ATPase was discovered in a commonly used preparation of ATP made by the Sigma Chemical Company, St. Louis, Mo. (Sigma grade ATP). As measured by a reliable and widely used assay system in which phosphate liberation in measured colorimetrically, Na-K-ATPase activity in the rat kidney, small intestine and colon was about 50% lower then Sigma grade ATP was used as substrate as compared to another Sigma Chemical Company product II ATP. Mg-ATPase and adenylate cyclase assays were unaffected by substituting Sigma grade for grade II ATP. The inhibitor of Na-K-ATPase could not be identified. Sigma grade ATP probably should not be used when measuring the activity of Na-K-ATPase in the rat kidney, small intestine, or colon.


A thermal method for the determination of tissue blood flow. An isothermal flowmeter for the determination of local tissue blood flow is described. Flow is determined by the measurement of the thermal conductivity of the tissue in the vicinity of a heated thermistor maintained at a fixed temperature difference above a reference thermistor. Direct heating of the thermistor is utilized to eliminate the need for specially constructed indirectly heated thermistors. This design results in a device with a voltage output directly proportional to tissue thermal conductivity and to tissue blood flow. The device is shown to be adequate for the qualitative measurement of myocardial blood flow under various situations. Construction is simplified and the size of the circuit reduced by the use of readily available integrated circuits.


Automated method for the determination of niacin and niacinamide in cereal products: collaborative study. In a collaborative study, an automated method for the determination of niacin and niacinamide in cereal products was compared with the official final action microbiological (43.121-43.125) and chemical (43.044-43.046) methods. Ten samples of cereal products, including enriched flour, yeast-leavened baked products, fortified breakfast cereals, and baked pet food products, were submitted to 14 laboratories. Nine laboratories reported values by the automated method, 6 reported values by the microbiological method, and 7 reported values by the chemical method. The results from the microbiological method were not subjected to analysis of variance because of the unusually large between-laboratory variation. The between-laboratory coefficients of variation for the automated and chemical methods were 10.90 and 10.18%, on the basis of results from 7 and 4 laboratories, respectively. There was no significant (p greater than 0.05) difference between methods when results from the 4 laboratories who used both methods were compared. The automated chemical method has been adopted as official first action.


Changes in glucosamine and galactosamine levels during conidial germination in Neurospora crassa. The levels of glucosamine and galactosamine were determined in conidia, germinating conidia, and vegetative mycelia of Neurospora crassa. In the vegetative mycelia about 90% of the amino sugars were shown to be components of the cell wall. The remaining 10% of the amino sugars were tentatively identified as the nucleotide sugars uridine diphospho-2-acetamido-2-deoxy-D-glucose and uridine diphospho-2-acetamido-2-deoxy-D-galactose. Conidia and vegetative mycelia contained about the same levels of glucosamine. During the first 9 h after the initiation of germination, the total glucosamine content had increased 3.1-fold, whereas the residual dry weight of the culture had increased 7.7-fold. This led to a drop in the glucosamine concentration from 100 mumol/g of residual dry weight to 42 mumol/g. During this time, all of the conidia had germinated and the surface area of the new germ tubes had increased to 10 times that of the conidia. Either germ tubes were initially produced without glucosamine-containing polymers, or these polymers (probably chitin) were deposited only at low densities in the germ tube cell walls. The chitin precursor uridine diphospho-2-acetamido-2-deoxy-D-glucose was present at all times during conidial germination. Conida contained very low levels of galactosamine. During germination, galactosamine could not be detected until the culture had reached a cell density of about 0.6 mg of residual dry weight per ml of growth medium. This was observed regardless of the time required to reach this cell density or the fold increase in dry weight. The accumulation of galactosamine-containing polymers does not appear to be necessary for germ tube formation. The levels of soluble galactosamine (uridine diphospho-2-actamido-2-deoxy-D-galatose) were very low in conidia and increased during germination at the same time that galactosamine appeared in the cellular polymers. In addition, under certain culture conditions, the appearance of galactosamine and the increase in the glucosamine concentration occurred simultaneously.


Developmental control of glucosamine and galactosamine levels during conidation in Neurospora crassa. The glucosamine and galactosamine content of mycelia was measured in cultures of Neurospora crassa grown on the surface of dialysis membranes. The glucosamine content was relatively constant throughout the different regions of the mycelial mat. The galactosamine content, however, was always lower in the growing-front region of the mycelial mat than in the older regions. At most, only low levels of galactosamine were necessary for the formation of hyphae at the growing front of a mycelial mat. Thus, galactosamine-containing polymers cannot be a major shape-determining component of the cell walls of these hyphae in Neurospora. The effect of conidiation on the amino sugar content was determined by using the bd (band) strain of N. crassa. When grown on the surface of dialysis membranes, this strain rhythmically produced regions of conidiating and non-conidiating growth. With this strain, it was concluded that conidiation did not affect the amino sugar levels. Since conidia that contained only very low levels of galactosamine were produced from regions of the mycelial mat that contained much higher levels of this amino sugar, there must be some mechanism of spatial differentiation that prevented the accumulation of galactosamine-containing polymers in conidia.


Characterization of the glucose transport systems in Neurospora crassa sl. Neurospora crassa sl, a mutant that lacks a rigid cell wall, exhibits transport systems for glucose similar to those of wild-type strain 1A. When the orgnism is grown in a medium containing 50 mM glucose as the carbon source, glucose is transported primarily by a glucose-facilitated diffusion system (GluI). When it is grown in a medium with little or no glucose present, a glucose active transport system (Glu II) is expressed. Both of these systems are similar kinetically to those in the wild type. Significant differences do exist between strains sl and 1A with respect to genetic regulation of the glucose active transport system.


Suppression of spectinomycin resistance in a mutant of Escherichia coli K-12. A mutant of Escherichia coli has been isolated which exhibits partial suppression of spectinomycin resistance. The site of mutation is in the streptomycin (strA) region and is closely linked to the spcA gene. However, this gene, which we propose to call mod, is phenotypically distinguishable from both the neomycin-kanamycin (nek) and the ribosomal ambiguity gene (ram). The relative gene order is mod spcA strA. In a cell-free protein synthesizing system, altered ribosomes appear to be responsible for the suppression of spectinomycin resistance caused by mod.


Regulation of exocellular proteases in Neurospora crassa: metabolic requirements of the process. To induce exocellular proteolytic enzyme from carbon-starved exponential-phase cells of Neurospora crassa, both a protein substrate and an activating protease of certain specific properties must be present at the same time. The cells must be capable of protein synthesis, since cycloheximide inhibits the process, but cell growth, as determined by increase in cell mass, does not appear to be required. Both soluble (bovine serum albumin, myoglobin) and insoluble protein substrates (collagen, corn zein) will affect protease induction, although certain soluble, globular proteins (egg white globulin, bovine gamma globulin) will not. In most cases, rates of protease induction are proportional to protein concentration, regardless of the nature of the inducing protein. All activating proteases capable of affecting induction in a manner similar to that of N. crassa exocellular protease were of bacterial origin and were exoproteases. Mammalian proteases and peptidases had little or no effect on the induction process.


PfkB and pfkC loci of Escherichia coli.

PfkB and pfkC loci of Escherichia coli. Mutants lacking Escherichia coli phosphofructokinase (pfkA, 78 min) are suppressed by the unlinked pfkB1 mutation, which restores some enzyme activity (Morrissey and Fraenkel, 1972). We here describe a secondary mutation at pfkB, "PFKB-," which abolishes the suppression as well as the low residual activity of unsuppressed pfkA mutants. pfkB is at about 33 min. with the gene order groD-pps-pheS-pfkB. A positive selection was found that yielded both the pfkB-mutations and a new similar mutation, pfkC-. pfkC is an early marker in Hfr HL16(ca. 50 to 55 min). Some pfkC-, but no pfkB-, mutations were amber. A temperature-sensitive pfkB- was also obtained. Strains carrying pfkB- or pfkC-, but wild type at pfkA, were not markedly affected in growth on sugars. A new search for suppressors such as pfkB1 gave five independent candidates, all of which suppressed both pfkA1 and pfkA2 and occurred in the pfkB region; none occurred at pfkC. Neither the pfkB nor the pfkC loci have assigned functions. It is likely that they are somehow involved in expression of phosphofructokinase activity 2 (Fraenkel, Kotlarz, and Buc, 1973).


PfkA locus of Escherichia coli.

PfkA locus of Escherichia coli. pfkA was know, on the basis of three mutants, as the likely locus of phosphofructokinase in Escherichia coli, and the unlinked pfkB1 mutation suppressed these mutations by restoring some enzyme activity (Morrissey and Fraenkel, 1972). We now report a new search for the complete inactivation of pfkA (e.g., by deletion or amber mutation), done to assess whether the pfkB1 suppression is by an independent enzyme, phosphofructokinase activity 2 (Fraenkel, Kotlarz, and Buc, 1973). Ten new phosphofructokinase mutants all were at pfkA, rather than at pfkB or pfkC. One of them (pfkA9) gave temperature-sensitive reverants with heat-labile enzyme. Another (pfkA11) proved genetically to be a nonsense mutation, but showed no restored activity when suppressed by supF. However, even unsuppressed it was found to contain an enzyme related to phosphofructokinase activity 1 kinetically (more allosteric), physically (almot identical subunit), and antigenically. All the pfkA mutants apparently contained cross-reacting material to activity 1. All (including pfkA11) were suppressed by the pfkB1 mutation. Several results support the idea that pfkA is the structural gene for the main phosphofructokinase of E. coli (activity 1), but that there is some restriction to its complete inactivation.


Scanning electron microscopy of surface and internal features of developing perithecia of Neurospora crassa. Stages in the development of perithecia of Neurospora crassa, designated by the time elapsed after crossing, were investigated with the scanning electron microscope, from protoperithecia through perithecia. The usual examination of external features of whole specimens with this instrument was augmented by a freeze-fracture technique which allowed the viewing of development internally as well. Rapid increases in perithecial size soon after crossing were followed by the appearance, in section, of a centrum, at first undifferentiated but subsequently developing ascogenous hyphae. The perithecial beak appeared as a compact mass easily distinguishable in whole specimens from the surrounding hyphae by means of texture as well as shape. Two ascospores were photographed during emergence from an ostiole, but ostioles were found more frequently closed than open.


Microbial transformation of artificial estrogens of the allenolic group. When 2,2-dimethyl 3-(2'-naphthyl 6'-hydroxy) pentanoic acid, an artificial estrogen of the allenolic acid group, was added to an exponential-phase growth culture of Neurospora crassa (in Horowitz medium), it was transformed into its hydroxylated derivative, 2,2-dimethyl 3-(2'-naphthyl 4',6'-dihydroxy)pentaoic acid. To study this transformation, radioactive 2-methyl-[2-14-C=A1methyl 3-(2'-naphthyl 6'-hydroxy) pentanoic acid has been prepared. The rate of metabolism of allenolic acids varies in the same way as their estrogenic activity.


Anacystis nidulans mutants resistant to aromatic amino acid analogues. Three classes of mutants of Anacystis nidulans were selected on the basis of resistance to fluorophenylalanine and 2-amino-3-phenylbutanoic acid. The most frequent type exhibited DAHP synthetase (7-phospho-2-keto-3-deoxy-D-arabino-heptonate-D-erythrose-4-phosphate-lyase [pyruvate phosphorylating], EC 4.1.2.15) activity identical to that of the parental strain. The second type was characterized by extremely low levels of the activity. The third type had a DAHP synthetase showing decreased sensitivity to inhibition by L-tyrosine. The enzyme was purified 140-fold from wild-type and feedback-insensitive strains, and the kinetics of the reaction was examined. The activity of the wild-type enzyme was inhibited 75% in the presence of 2.0 X 10-3 M tyrosine, and the altered enzyme was inhibited 10%. The following apparent constants were obtained from kinetic studies with partially purified wild-type enzyme: S0.5 for D-erythrose-4-phophate equal to 7.1 X 10-4 M; S0.5 for phosphoenolpyruvate equal to 1.4 X 10-4 M. Inhibition by tyrosine was mixed with respect to binding of both D-erythrose-4-phosphate and phosphoenolpyruvate. In addition, tyrosine promoted cooperative interactions in the binding of phosphoenolpyruvate. For the altered enzyme the following apparent constants were obtained: S0.5 for D-erythrose-4-phosphate equal to 7.1 X 10-4 M; S0.5 for phosphoenolpyruvate equal to 2.9 X 10-4 M. Inhibition by tyrosine was mixed with respect to D-erythrose-4-phosphate and competitive with respect to phosphoenolpyruvate. Tyrosine did not promote cooperative effects in the binding of phosphoenolpyruvate to the altered enzyme.


Size restriction on utilization of peptides by amino acid auxotrophs of Neurospora crassa. Growth of an amino acid auxotroph of Neurospora crassa on oligopeptides is shown to occur by extracellular hydrolysis, with subsequent utilization of monomer amino acid residues, and by transport of peptides. Peptides with a hydrodynamic volume greater than that of trileucine are not transported, and this lack of transport is shown to be due to restriction by the oligopeptide transport system rather than the cell wall.


Enzymatic and non-enzymatic reduction of nitrite by extracts of Neurospora crassa. Two activites causing nitrite disappearance are found in extracts of Neurospora; one, inducible by nitrate or nitrite and present only in nitrite-utilizing strains, catalyze the stoichiometric reduction of nitrite to ammonia; the other, present in all strains under all conditions, causes the disappearance of nitrite to something other than ammonia. The latter activity has a molecular weight of about 600 and may contain an oligopeptide, a metal, and an SH group(s). It has no known physiological function.


Effects of sodium and potassium ions on the elementary steps in the reaction of Na+-K+-dependent ATPase1. The effects of Na+ and K+ ions on the elementary steps in the reaction of Na+-K+-dependent ATPase (EC 3.6.1.3) were investigated in 0.5-600mM NaCL and 0-10mM KCL, at a fixed concentration (1mM) OF MgCL2, AT PH 8.5 and at 15 degrees. The data were analyzed on the basis of the reaction mechanism in which a phosphorylated intermediate, E ADP P (abbreviated as EP), is formed via two kinds of enzyme-substrate comples, E1ATP and E2ATP, and EP is in equilibrium with E2ATP, and is hydrolyzed to produce P1 and ADP. The following results were obtained: 1. The rate od E2ATP-formation, vf, increased with increase in the Na+ concentration, reached a maximum level, and then decreased with further increase in the Na+ concentration at various K+ concentrations. The value of vf was given as (see article). 2. The reciprocal of the equilibrium constants, K2, of the step E1ATPEQUILIBRIUM E ADP P in the presence of low concentrations of Na+ was larger than that in the presence of high concrntrations of Na+, indicating that the equilibrium shifted markedly toward E2ATP at low concentrations of Na+. The relation of K3 with Na concentration was rather complicated on varying the concentration of K+. However, generally speaking, it increased with increase in the K+ concentration. 3. The decomposition of EP was markedly activated by even low concentrations of K+, and inhibited by high concentrations of Na+. The inhibition by Na+ was partially suppressed by K+. The rate constant of EP-decomposition, vo/(EP), was given by (see article) where (vo/(EP) K+EQUALS0  was the value of vo/[EP] in the absence of K+.


Properties of the conversion of an enzyme-ATP complex to a phosphorylated intermediate in the reaction of Na+-K+-dependent ATPase1. Previously, we proposed the following reaction machanism for the transport ATPase (EC 3.6.1.3) reaction in the presence of high concentrations of Mg2+ and Na+:(see article). Some kinetic and thermodynamic properties of steps 3 and 4 were investigated, and the following results were obtained. 1. When the reaction was started by adding ATP to the enzyme in the presence of 50 mM Na+ and 0.5 mM K+ or in the presence of 50mM Na+ and 0.5mM Rb+, the amount of E ADP P increased with time and maintained a constant level after reaching a maximum. We could not observe the initial burst of EP formation, which was observed by Post er al. in the presence of 8 mM Na+ and 0.01 mM Rb+. 2. The existence of quasi-equilibrium between E2ATP and E ADP P in the presence of low concentrations of Na+ was suggested by the fact that the values of the reciprocal of the equilibrium constant, K3 of step 3 obtained by the following three methods were almost the same. a) The value of 1+K3 was estimated from the ratio of vo/[EP] to kd, where vo is the rate of ATP hydrolysis in the steady state, [EP] the concentration of EP, and kd the first-order rate constant of EP disappearance after stopping EP formation. b) This value was also calculated from the ratio of the amount of P1 liberated to that of decrease in EP after stopping EP formation. c) The value of K3 was also calculated from the initial rapid decrease in EP on adding K+ and EDTA, assuming that the rapid decrease was due to a shift of the equilibrium toward E2ATP on adding K+. For example, the value of K3 with 10mM NaCL and 0.5mM KCL was 7--11. Although ATP formation due to a shift of the equilibrium toward E2ATP by a K+ jump in the presence of a low concentration of Na+ was observed at 0 degrees, the amount of ATP formed by a K+ jump at 15 degrees was less than the value expected from the shift of the equilibrium. 3. The values of delta H degrees  and delta S degrees of step 3 were estimated in the presence of a sufficient amount of Na+ and in the absence of K+. They were +4--+5 kcal mole minus 1 and +15--+16 entropy units mole minus1, respectively. On the basis of kinetic studies of the elementary steps and the overall reaction of Na+-K+-dependent ATPase [EC 3.6.1.3], we (1--4) showed that a phosphorylated intermediate, EP, is formed via two kinds of enzyme-substrate complex, E1ATP and E2ATP, that the EP is in K+-dependent quasi-equilibrium with E2ATP, and that in the presence of high concentration of Mg2+, EP is in a high-energy state and contains bound ADP, E ADP P.(see article).


Desensitization of substrate inhibition of acto-H-meromyosin ATPase by treatment of H-meromyosin with rho-chloromercuribenzoate. Relation between the extent of desensitization and the amount of bound rho-chloromercuribenzoate1. H-Meromyosin (CMB leads to betaME-H-meromyosin) was prepared by tryptic digestion of myosin, which had been treated with CMB bound to H-meromyosin and the extent of desensitization of the substrate inhibition of acto-H-meromyosin ATPase [EC 3.6.1.3.] was investigated. Both the dissociation of acto-H-meromyosin induced by ATP and substrate inhibition decreased with increase in the amount of bound CMB to a minimum value at about 1 mole of CMB bound per mole of H-meromyosin. The substrate inhibition of acto-H-meromyosin ATPase was restored to the original level by complete removal of the bound CMB by further treatment of CMB leads to beta ME-H-meromyosin with a large excess of beta-mercaptoethanol. The dissociation constant of acto-H-meromyosin in the presence of ATP decreased markedly on modification with CMB, while the maximum ATPase activity ar a sufficiently high concentration of F-actin remained essentially unchanged. Acto-H-meromyosin was reconstituted from F-actin and CMB LEADS TO beta ME-H-meromyosin, containing less than the stoichiometric amount of bound CMB. Its ATPase activity and the extent of dissociation of acto-H-meromyosin induced by ATP were explained as those of a mixture of unmodified H-meromyosin and CMB leads to beta ME-H-meromyosin containing 1 mole of CMB per mole of H-meromyosin. Half of the light chains (g2), with a molecular weight of 18,000, were removed from myosin by treatment with CMB and beta-mercaptoethanol. After this treatment, on further incubation of the myosin with a large excess of beta-mercaptoethanol, the myosin contained only half of the g2, but the substrate inhibition of acto-H-meromyosin ATPase was restored completely. The initial burst of P1 liberation and the EDTA-ATPase activity decreased to almost zero on specific modification of the SH1-groups with NEM, while the initial burst decreased to some extent and the EDTA-ATPase activity to 50% of the original value on binding of 1 mole CMB per mole of H-meromyosin. The actomyosin-type of ATPase activity was strongly inhibited by modification with CMB. The extent of the dissociation of acto-H-meromyosin induced by ATP was unaffected by modification with NEM, while it decreased on further treatment of NEM-myosin with CMB FOLLOWED BY BETA-MERCAPTOETHANOL.


Cytoplasmic membrane vesicles of Escherichia coli.  A simple method for preparing the cytoplasmic and outer membranes. A simple preparative method is described for isolation of the cytoplasmic and outer membranes from E. coli. The characteristics of both membrane fractions were studied chemically, biologically, and morphologically. Spheroplasts of E. coli K-12 strain W3092, prepared by treating cells with EDTA-lysozyme [EC 3.2.1.17], were disrupted in a French press. The crude membrane fraction was washed with 3 mM EDTA-10% (w/v) sucrose, pH 7.2, and the cytoplasmic membranes and outer membranes were separated by sucrose isopycnic density gradient centrifugation. The crude membrane fraction contained approximately 10% of the protein of the whole cells, 0.3% of the DNA, 0.7% of the RNA, 0.3% of the peptidoglycan, and about 30% of the lipopolysaccharide. The cytoplasmic membrane fraction was rich in phospholipid, while the outer membrane fraction contained much lipopolysaccharide and carbohydrate; the relative contents of lipopolysaccharide and carbohydrate per mg protein in the cytoplasmic membrane fraction were 12 and 40%, respectively, of the contents in the outer membrane fraction. Cytochrome b1, NADH oxidase, D-lactate dehydrogenase [EC 1.1.1.28], succinate dehydrogenase [EC 1.3.99.1], ATPase [EC 3.5.1.3], and activity for concentrative uptake of proline were found to be localized mainly in the cytoplasmic membranes; their specific activities in the outer membrane fraction were 1.5 to 3% of those in the cytoplasmic membrane fraction. In contrast, a phospholipase A appeared to be localized mainly in the outer membranes and its specific activity in the cytoplasmic membrane fraction was only 5% of that in the outer membrane fraction. The cytoplasmic and outer membrane fractions both appeared homogeneous in size and shape and show vesicular structures by electron microscopy. The advantages of this method for large scale preparation of the cytoplasmic and outer membrane fractions are discussed.


Prevention of freeze denaturation of carp actomyosin by sodium glutamate. 1) Denaturation of carp actomyosin during storage at -20 degrees was studied with particular interest in the cryoprotective effect of sodium glutamate, the most cryoprotective of the compounds tested previously.  2) Storage with glutamate prevented the rapid decrease in solubility, viscosity, and ATPase (EC 3.6.1.3)activity of actomyosin during storage.  Ultracentrifugal studies suggested that aggregation occurred in the frozen state without glutamate, but that added glutamate prevented aggregation or denaturation. 3) Electron  microscopy showed that the original actomyosin consisted of long filaments with typical "arrowhead" structures, and that these decomposed into small fragments and sticked with globular portions, forming loosely packed aggregates during storage without glutamate.  On storage with glutamate, the filaments were well preserved, and their fine structure was clearer than that of the original sample.  4) Preparations of actomyosin extracted with 10 mM glutamate were of better quality and their ultrastructure and physicochemical and biochemical properties showed increased stability on freezing.  5) Freeze-denaturation seems to involve complicated aggregation with transconformation of proteins besides the side-to-side aggregation discussed previously.


Standard free energy changes for formation of various intermediates in the reaction of H-meromyosin ATPase. Two reaction intermediates of H-meromyosin (HMM) ATPase [EC 3.6.1.3], E2AT32P, and (see article), were formed by mixing excess HMM with AT32P. Then a large excess of unlabelled ATP was added, and the amount of AT32P liberated from E2AT32P was measured as the difference between the total amount of AT32P in the reaction mixture and the amount of AT32P bound to HMM, obtained by filtering the mixture after adding charcoal to adsorb nucleotides (charcoal-filtration method). The amount of free AT32P was also measured as the amount of glucose-6-32P formed within 15 sec after adding large excesses of hexokinase [EC 2.7.1.1] and glucose to the reaction mixture. The rate constant, k-2, for the step E2ATP yields E plus ATP was calculated at various KCl concentrations from the time-course of liberation of AT32P. The intermediate, (see article), was formed by mixing HMM with AT32P in a molar ratio of 1:2, and the rate constant, k-6, for the step (see article) was also determined by the same procedures used for k-2. In 0.5 M KCl and 2 mM MgCl2 at pH 7.8 and 0 degrees, k-2 and k-6 were 0.002 sec-1 and 0.1 sec-1 or more, respectively. From the rate constants determined in this work and the rate and equilibrium constants which we reported previously, the standard free energy changes (kcal/mole) for formation of various reaction intermediates in the reaction of HMM ATPase in 0.5 M KCl and 2 mM MgCl2 at pH 7.8 and 0 degrees were calculated to be as follows: (see article).


Influence of aurovertin on mitochondrial ATPase activity. Investigations have been made of the kinetic effects of the antibiotic aurovertin on the ATPase and ITPase activity of isolated rat liver mitochondrial ATPase. Unusual patterns of inhibition, decreasing slope, and increasing y-intercept values of double reciprocal plots, were observed with Mg-ATP as the substrate under various conditions. Under specified conditions, aurovertin stimulated hydrolysis of MgATP. The inhibition of MgITP hydrolysis was uncompetitive. Aurovertin eliminated the HCO3-minus stimulation of MgATP hydrolysis. The implications of these findings for the mechanism of mitochondrial ATPase are briefly discussed.


Native (Na-+ + K-+)-dependent adenosine triphosphatase has two trypsin-sensitive sites. Sodium and potassium adenosine triphosphatase ((Na + K)-ATPase) consists of two polypeptides, a large molecular weight polypeptide (MW 84,000 to 102,000) and a sialoglycoprotein (MW 35,000 to 57,000). Trypsin treatment of this complex selectively cleaves the large polypeptide into two fragments with molecular weights of 62,000 and 43,000. Simultaneously with the appearance of these fragments, (Na + K)-APTase activity is destroyed. Trypsin treatment of phosphorylated enzyme shows that he 43,000 molecular weight fragment is phosphorylated. If (Na + K)-ATPase is digested with trypsin in the presence of ATP, a 90,000 molecular weight fragment is produced. Disappearance of the large polypeptide, and loss of ATPase activity parallel the production of this fragment. Addition of strophanthidin to this mixture significantly lowers the amount of the 90,000 molecular weight fragment produced. Experiments on (Na + K)-ATPase of the red cell membrane suggest that trypsin is cleaving (Na + K)-ATPase at the interior surface of the plasma membrane.


Primary structure of human fibrinogen and fibrin. Isolation and partial characterization of chains of fragment D. Fragment D has been isolated as an apparently single molecular weight species (molecular weight about 100,000) from plasmin digests of humman fibrinogen, using a combination of affinity chromatography on insolubilized "fibrin monomer" and gel filtration. This fragment consists of three chains with molecular weights of 15,000 (Dbeta), 42,500 (Dgamma1) or 39,500 (Dgamma2), and 14,000 (Dalpha) held together by disulfide bonds. The S-carboxymethyl derivatives of the chains have been separated by gel filtration and ion exchange chromatography, and their identity has been confirmed by peptide mapping and immunological analysis. The chain with a molecular weight of 45,000 is a fragment of the Bbeta chain of fibrinogen. The chain derived from the gamma chain of fibrinogen occurred in two molecular forms having molecular weight 42,500 and 39,500. The chain derivative with molecular weight 14,000 is most likely derived from the Aalpha chain of fibrinogen. The chains were characterized by NH2-terminal sequence analysis, amino acid composition, and carbohydrate staining. The two molecular analysis, amino acid composition, and carbohydrate staining. The two molecular forms of the gamma chain appeared to be identical except for an NH2-terminal peptide extension of 23 amino acid residues in the longer chain. The latter has sequences in common with the COOH-terminal part of the gamma chain of the NH2-terminal disulfide knot (BROMBACK, B., BRONDAHL, N. J., HESSEL, B., IWANAGA, S., and WALLEN, P. (1973) J. Biol. Chem. 248, 5806-5820); its NH2-terminal residue being Ala-63 of the gamma chain of fibrinogen.


Purification and characterization of catabolic dehydroquinase, an enzyme in the inducible quinic acid catabolic pathway of Neurospora crassa. Catabolic dehydroquinase which functions in the inducible quinic acid catabolic pathway in Neurospora crassa has been purified 8000-fold. The enzyme was purified by two methods. One used heat denaturation of contaminating proteins; the other used antibody affinity chromatography. The preparations obtained by these two methods were identical by all criteria. The purified enzyme is extremely resistant to thermal denaturation as well as denaturation 0y urea and guanidine hydrochloride at 25 degrees. It is irreversibly inactivated, although not efficiently dissociated, by sodium dodecyl sulfate and guanidine hydrochloride at 55 degrees. At pH 3.0, the enzyme is reversibly dissociated into inactive subunits. At high concentrations catabolic dehydroquinase aggregates into an inactive, high molecular weight complex. The native enzyme, which has a very high specific activity, has a molecular weight of approximately 220,000 and is composed of identical subunits of 8,000 to 12,000 molecular weight each. The native enzyme and the subunit are both asymmetric.


Na+ATPase of the mammalian erythrocyte membrane. Reversibility of phosphorylation at 0 degrees. When human erythrocyte membranes are phosphorylated with a very low concentration of [gamma-32P]ATP (0.02 muM) at 0 degrees, and then EDTA is added, rapid disappearance of the phosphoenzyme intermediate of Na+ATPase is observed. The initial rapid phase of phosphoenzyme disappearance is, for the most part, not associated with P1 release and its rate constant, kD, is severalfold greater than the ratio of Na+ATPase activity to phosphoenzyme intermediate, v:EP, at steady state. It is concluded that this rapid disappearance of phosphoenzyme is due to resynthesis of ATP via reversal of phosphorylation. In contrast, rapid reversal is not observed when excess nonradioactive ATP is added to reduce E32P formation, provided Mg2+ is present; however, K+ added with the ATP stimulates reversal. Rapid reversal following EDTA addition is unlikely also when higher ATP concentrations (greater than or equal to 10(-6) M) are used to phosphorylate the enzyme since, at higher ATP, kD congruent to v:EP. The results are compatible with the concept that the Na+ATPase enzyme is composed of two or more catalytic subunits, in which ATP at one catalytic site can regulate the reactivity at another site.


Isolation and characterization of proteoglycans from the swarm rat chondrosarcoma. Proteoglycan monomer (D1) and aggregate (A1) preparations were isolated from 4 M guanidinium chloride extracts of the Swarm rat chondrosarcoma. When EDTA, 6-aminohexanoic acid, and benzamidine were present in the solutions, the D1 preparation contained a single component (SO = 23 S), and the A1 preparation contained 30% monomer (SO = 23 S) and 70 percent aggregate (SO = 111 S). In the absence of EDTA, 6-aminohexanoic acid, and benzamidine, the A1 preparations contained only small proteoglycan fragments, indicating that extensive enzymatic degradation had occurred. The composition of the proteoglycan monomer was different from that of proteoglycan monomer preparations from normal hyaline cartilages in that it did not contain keratan sulfate and chondroitin 6-sulfate; only chondroitin 4-sulfate was found. The A1 preparation from the chondrosarcoma contained only one link protein, which was like the smaller (molecular weight of 40,000) of the two link proteins present in A1 preparations from bovine nasal cartilage. When the A1 preparation from the chondrosarcoma was treated with chondroitinase ABC and trypsin and the digest was chromatographed on Sepharose 2B, a complex was isolated which contained the link protein and the segments of the protein core from the hyaluronic acid-binding region of the proteoglycan molecules.


The substructure of heavy meromyosin. The effect of Ca2+ and Mg2+ on the tryptic fragmentation of heavy meromyosin. Heavy meromyosin, obtained by tryptic digestion of myosin, containing two main polypeptides whose masses were estimated as 81,000 and 74,000 dlatons from Na dodecyl-SO4 polyacrylamide gel electrophoresis, was further digested with trypsin. The Ca2+-activated ATPase activity remainded unchanged and the K+-EDTA activity increased while various smaller fragments were formed. The formation of some of these fragments is affected by Ca2+ or Mg2+ as first shown by Bálint et al. (Bálint, M., Schaefer, A., Biro, N. A., Menczel, L., AND Fejes, E. (1971) J. Physiol. Chem. Phys. 3, 455). On the basis of the time course of the appearance of fragments the following relationship emerges: see article. The 64K leads to 60K step is inhibited by divalent cations, while the breakdown of the 74K fragment is accelerated. The effect of Ca2+ was maximal at 0 similar to 0.1 muM, that of Mg2+ at 10 muM. The original light chains of myosin are not present in the heavy meromyosin serving as the starting material, but peptide material appears on electrophoresis in positions starting material, but peptide material appears on electrophoresis in positions where the light chains would be found. The fragments marked by an asterisk are considered to ba alpha-helical on the basis of their solubility at low ionic strength after precipitation with ethanol (Bálint et al.). The fact that alpha helical fragments are derived from the 60,000-dalton fragment indicateds that it is adjacent to the light meromyosin in the intact myosin while the 74,000- dalton fragment would be part of heavy meromysoin subfragment 1. Chromatography of Sephadex G-200 separates fractions with ATPase activity corresponding to heavy meromyosin and heavy meromyosin subfragment 1. Electrophoresis of these Sephadex fractions suggests that the main peptide constituting heavy meromysoin subfragment 1 is connected by noncobalent forces to a portion of the rod that is not immediately adjacent to it in the primary sequence. The significance of this finding is discussed in terms of the flexibility of the myosin head.


[Rupture of the spleen in patients with multiple injuries. Current diagnostic methods. Apropos of 60 cases]. The authors report 60 cases of severe multiple injuries and study present methods permitting a rapid diagnosis of latent rupture of the spleen. Purely clinical methods are today supplemented by paraclinical investigations. e.g. --intra-abdominal catheterisation; --peritoneal wash-out; --laparoscopy; --and, above all, arteriography. The authors use these methods and weigh the advantages and disadvantages of each.


Exaggerated growth hormone response to arginine infusion in Huntington's disease. Growth hormone regulation was studied in 10 patients with Huntington's disease after intravenous administration of arginine. In 20 control subjects arginine infusion resulted in a rise of plasma growth hormone levels from a mean baseline value of 3.2+/-0.6 ng/ml to a peak level of 17.6+/-2.7 ng/ml at 60 min. Growth hormone rise in the majority of patients with Huntington's disease was clearly intact and significantly greater than normal in magnitude, increasing from the baseline level of 2.6+/-0.5 ng/ml to a peak level of 28.3+/-3.7 ng/ml at 60 min (P = less than 0.05). Carbohydrate tolerance of these patients was previously examined, and 4 with normal glucose tolerance and normal insulin responses to arginine infusion had growth hormone levels significantly higher than controls at 30 min. Six patients with impaired carbohydrate tolerance and exaggerated insulin responses to arginine had significantly higher growth hormone responses at 30 min and also at 60 min. Neuronal degeneration of several hypothalamic nuclei has been reported in Huntington's disease. The observations that growth hormone responds in an exaggerated fashion to stimulation by arginine infusion or falling glucose levels as previously described may be explained by intrahypothalamic dysfunction such as impairment of somatostatin secretion.


Early failure of Dacron prosthetic grafts. Crimped knitted Dacron grafts are commonly employed as arterial substitutes and in most instances have proven to be highly effective and relatively durable. Graft failures have occurred and have usually been associated with serious hemorrhage. Fabrication flows, intraoperative damage to the graft, or biodegradation have been suggested as causes of graft failure. In this study three aortic grafts failed, each apparently from a different cause and each presented in a different fashion. Although the exact cause of graft failure in these three cases could not be ascertained it appeared that one was due to a fabrication flow, one due to generalized loss of tensile strength, and the third due to deterioration of only a portion of the fraft. These data suggest that continued search for more suitable plastic prostheses is urgently needed;


Deoxyribonucleic acid strandedness. Partial characterization of the antigenic regions binding antibodies in lupus erythematosus serum. This study shows that tritiated thymidine labeled DNA prepared from mammalian cells by the Marmur technique is a pure preparation of nucleic acid that is composed essentially of two populations of molecules. One molecular population consists of primarily double-standed nucleic acid, while the other population is of double-stranded nucleic acid with significant single-stranded regions. The double-stranded DNA with single-stranded regions can, depending upon the length of the single strand, behave as "native" DNA or "denatured" DNA on methylated albumin kieselguhr (MAK) column chromatography, Using MAK chromatography we have separated the DNA into a saltelutable fraction composed of primarily double-stranded molecules and an alkaline-elutable fraction containing double-stranded nucleic acid with variable length, single-stranded regions. Endonuclease enzyme removal of the single-stranded regions from the alkaline fraction DNA yield nucleic acid that behaves identically to the salt elutable DNA. Exonuclease removal of the single-stranded regions suggests they are located primarily at the ends of the molecules. Our data show that the alkaline-elutable DNA differs from salt-elutable DNA only in that the former has significant single-stranded regions. Sera of patients with systemic lupus erythematosus (SLE) selected for anti-DNA by hemagglutination bind significantly less to the alkaline fraction DNA than the sale fraction DNA. This difference in binding clearly does not represent simply an affinity for double-stranded vs. single-stranded nucleic acid since the alkaline fraction DNA contains predominately double-stranded nucleic acid. A model for antibody-DNA binding is suggested from the present data and information contained in the literature.


Immune complexes in sera and synovial fluids of patients with rheumatoid arthritis. Radioimmunoassay with monocylonal rheumatoid factor. Evidence for the presence of immune complexes in blood, synovial fluid, and tisues of patients with rheumatoid arthritis (RA) includes low complement levels in blood and effusions, deposition of immunoreactants in tissues and vessel walls, precipitate formation after addition of monoclonal rheumatoid factor (mRF) to serum or synovial fluid. To quantitate immune complex-like material in RA patients, we developed a radioimmunoassay based on inhibition by test samples of the interaction of (125I)aggregated IgG (agg IgG) and mRF coupled to cellulose. This method could measure immune complexes of human antibody with hemocyanine prepared in vitro. The assay was not influenced by presence of polyclonal RF in test samples, nor by freezing and thawing. Normal levels of immune complex-like material in serum were less than 25 mug agg IgG EQ/ML. 12 of 51 RA sera examined (26%) contained more than 25 mug/ml. The presence of this material in RA sera was found to correlate with severity of disease, as measured by anatomical stage and functional class. There was an inverse correlation of the material with serum C4 level. Rheumatoid synovial fluids generally contained higher levels than serum, and five of 23 contained very much higher levels. The frequency of elevated levels of immune complex-like material in sera of patients with systemic lupus erythematosus (2 of 29) and with miscellaneous vasculitides (2 of 21 was much lower than in RA, suggesting that mRF exhibits a specificity for only certain kinds of immune complexes. The reason for this apparent specificity may explain such distinctive features of RA as the high frequency of polyclonal RF, the lack of immune complex nephritis, and the generally normal levels of serum complement.


Hodgkin's disease. An immunodepleting and immunosuppressive disorder. Irradiated leukocytes or mononuclear leukocytes, from 16 out of 30 patients with Hodgkin's disease and from one patient with the Sézary syndrome, stimulated in culture subnormal (3H)thymidine incorporation by allogeneic lymphocytes from normal individuals. This abnormality was not demonstrated in any of 30 other patients with non-Hodgkin's lymphomas. Subnormal mixed leukocyte culture reaction activation was caused by suppression of the mixed leukocyte reaction by patients' cells. Inhibition of the reaction by patient mononuclear leukocytes was corrected when adherent cells were removed or when protein synthesis was inhibited with cycloheximide. The inhibitory cells were probably lymphocytes since selective removal of phagocytic cells did not remove the inhibition by other patient mononuclear leukocytes. The presence in culture of as few as 2,500 granulocytes per mm3 also reduced responses when target cells were from patients with Hodgkin's disease. Patient cells no longer suppressed the mixed leukocyte reaction after patients entered clinical remission which suggests that suppression is a reversible, disease-related abnormality. Thus, the immune deficiency with advance Hodgkin's disease caused by ly lymphocyte depletion may be compounded by a relative excess of suppressor lymphocytes. The overall immunodeficiency may be further compounded by suppression of immune response by granulocytes at even physiologic concentrations.


Histochemical fiber typing and staining intensity in cat and rat muscles. In the gastrocnemius muscle of cat and rat, staining for oxidative enzymes differentiated three fiber types (A,B,C) and staining for adenosine triphosphate at pH 9.4 differentiated two fiber types (I, II) with a reliability of 90% and 98%, respectively. In cat 96% and in rat 90% of the fibers were typed identically after staining for nicotinamide adenine dinucleotidelinked lactic dehydrogenase (LDH) and succinic dehydrogenase (SDH). When differentiated by staining for LDH, A and B fibers were of type I. IN RAT, 80-90% OF ALL FIBERS WERE OF TYPE 22, COMPPRISING A, B and C fibers. Type I fibers stained for LDH intensely as did C fibers of type II, but stained intermediately for SDH. The degree of staining was measured by photometry. When fibers were stained for LDH, histograms of density showed three peaks corresponding to A, B and C fibers in cat, but only two peaks corresponding to A and C fibers in rat, In cat and rat, the densities of A, B and C fibers belonged to different populations. In soleus muscle of cat and rat stained for LDH, menadione-linked alpha-glycerophosphate dehydrogenase and adenosine triphosphatase at pH 9.4, the degree of staining differed from thatin any type of fiber in gastrocnemius muscle


Antibody-dependent cell-mediated cytotoxicity in humans. I. Characterization of the effector cell. Mononuclear cells from human peripheral blood are able to destroy alloantibody-coated target cells of a human lymphoblastoid line. The effector cell is a nonadherent, non-phagocytic, medium sized lymphoid cell. The cell is not detectable in early human fetuses. Its activity is destroyed by x-ray treatment in vitro but is not inhibited by anti-immunoglobulin sera. Its efficiency in antibody-dependent cell-mediated cytotoxicity is rapidly lost in culture and this loss is prevented by stimulation with allogeneic cells.


3H-uridine incorporation by small lymphocytes of tolerant rats: relationship to T and B lymphocytes. Tissue tolerance was induced in neonatal rats by the intravenous injection of bone marrow cells from adult allogeneic rat donors. After 6 to 8 weeks, lymphoid cells from rats in which tolerance had been induced were tested for mixed lymphocyte reactivity (MLR), 3H-uridine uptake, and the relationship of uridine incorporation to B and T lymphocytes. Lymph node (LN) and spleen (SPL) cells from the adult inoculated rats showed no reactivity in the MLR or normal lymphocyte transfer reaction (NLTRx), indicating that the animals were tolerant. After in vitro exposure to 3H-uridine, an abundance of small lymphocytes (SL) from these same tolerant rats were heavily labeled, in contrast to nontolerant controls, where relatively few SL were heavily labeled. In order to determine whether the heavily uridine-labeled cells were T cells or B cells, lymphoid cells from the LN and SPL of tolerant animals were exposed to either rabbit anti-AKR brain serum or rabbit anti-rat Ig conjugated with ferritin. The results showed that the heavily uridine-labeled SL of the tolerant rats were mainly Ig-positive cells.


Immunogeneic analysis of H-2 mutations. II. Cellular immunity to the H-2DA mutation. The H-2da haplotype was derived from the H-2d haplotype by a mutation localized to the D end of the H-2 complex. Coculture of H-2d and H-2da spleen cells gives rise to bidirectional MLR. However, the H-2d anti-H-2da response is much stronger than that of H-2da anti-H-2d. Both haplotypes give rise to reciprocal CML. B10.D2(R103) strain spleen cells, which differ only at the D end of the H-2 complex from the H-2d haplotype, kill H-2da target cells in CML when sensitized to H-2d stimulators and vice versa. Therefore, both the mutant and strain of origin share a D end CML specificity. H-2d and H-2da reject skin grafts in both directions, although some H-2d grafts show prolonged acceptance on H-2da recipients. These data are consistent with a mutation in the D end of the H-2d haplotype resulting in gain-loss of an antigen(s) that gives rise to reciprocal MLR, CML, and skin graft rejection. Further, the mutant can be distinguished from the strain of origin on the basis of the strength of immune response in MLR.


Immunogenetic analysis of H-2 mutations. III. Genetic mapping and involvement in immune reactions of the H-2ka mutation. Mutation M523 (H-2ka) occurred spontaneously in strain CBA/CaLacSto and was discovered during routine skin graft testing for genetic homogeneity. By linkage and complementation tests, the mutation was previously mapped in the K end of the H-2 complex. We demonstrate that the mutation occurred in the K region, without affecting the I region in the K end of the complex. The mutant antigens cause rejection of skin grafts, stimulate cells in mixed lymphocyte culture, and function as stimulators as well as targets in cell-mediated lymphocytotoxicity. Yet, they are serologically indistinguishable from the antigens of the original strain and do not induce formation of humoral antibodies upon immunization of the CBA strain. Together with the results obtained on testing of other H-2 mutants, the data strongly support the notion that classical H-2 antigens (i.e., products of the H-2K and H-2D loci) can function as lymphocyte-stimulating determinants, and that I-region differences are not required for the induction of strong cell-mediated lymphocytotoxicity.


The circulating life span, immunocompetence and 3H-uridine uptake of small lymphocytes from thymus-grafted, neonatally thymectomized rats. By 7 weeks post-grafting, the number of small lymphocytes in the thoracic duct lymph (TDL) and blood of the thymus-grafted neonatally thymectomized adult rats had increased to 60% of the number of cells in sham controls, or 2-1/2 times thymectomized control values. This increasing consisted almost exclusively of long-lived, recirculating small lymphocytes and corresponded to a 60% recovery of cellular immunocompetence as measured by the mixed lymphocyte reaction (MLR). Associated with the return of cellular immunocompetence was an increased incorporation of 3H-uridine by the small lymphocytes. Cells from thymectomized animals grafted with lymph node fragments demonstrated no significant increase in lymphocyte numbers nor was there a return of immunocompetence as compared to thymectomized controls.


Modulation of HL-A antigens by anti-HLA antiserum: effects on the cytotoxicity assay and mixed leukocyte reaction. Modulation of human leukocyte antigen (HLA) was attempted by treating leukocytes with specific anti-HLA antiserum or by their passage through columns coated with anti-HLA or a double layer of HLA-anti-HLA. The modulated cells were resistant to the cytotoxic effects of the anti-HLA, and they were poor stimulators and good responders to allogeneic cells in the unidirectional mixed leukocyte reaction. Modulated cells regained their HLA 16 hr after modulation if kept in cell suspension alone. The proliferative responses of modulated cells to mitogens were as good as non-modulated cells, indicating that modulation was probably not caused by depletion of lymphoid cells. Supernatants of modulated cells that were incubated overnight or preformed HLA-anti-HLA complexes were capable of suppressing and enhancing the MLR of specific cells depending on the dose used. The similarities of modulation of HLA to other lymphocyte receptors and the limitation of application of the modulation phenomenon to transplantation of allogeneic cells are discussed.


Subpopulations of human thymus cells differing in their capacity to form stable E-rosettes and in their immunologic reactivity. The majority of human thymus cells from young donors form stable E-rosettes with sheep red blood cells (SRBC) that do not distintegrate after prolonged incubation at 37 degrees C. With advancing age the proportion of thymus cells forming such rosettes decreases gradually. The thymus of a patient receiving prednisone treatment was found to contain only a few cells that formed stable E-rosettes. The minor population of thymus cells that fails to form stable E-rosettes (non-rosetting or NR cells) was isolated and tested for its cell surface markers and immunologic reactivity in vitro. Most of the NR-cells were capable of forming regular E-rosettes with SRBC at room temperature. Like the majority of human thymus cells they were sensitive to the cytotoxic effect of normal constituted less than 0.2% of the original thymus cell suspensions, but about 1 to 3% of the NR-population. Thymus cells from donors over the age of 36 and from a prednisone-treated child responded in vitro to stimulation with either phytohemagglutinin (PHA) or concanavalin A (Con A). Unfractionated thymus cells from children up to the age of 14 failed to react to either PHA or Con A, but their NR-population responded vigorously to both lectins. In contrast to unfractionated thymus cell suspensions from children, the NR fraction showed a significant reactivity in mixed lymphocyte cultures with mitomycin-C treated allogeneic lymphocytes. It is concluded that like the thymus of other species, the human thymus contains a minor population of cortisone-resistant cells endowed with many of the immunologic properties characteristic for periperal T lymphocytes.


A passive agglutination method using collagen-coated tanned sheep erythrocytes to demonstrate collagen-glycosaminoglycan interaction. A haemagglutination method originally designed to detect and measure antibody has been shown to be capable of monitoring interactions of glycosaminoglycans and collagen in vitro under physiological conditions of pH and ionic strength. In this system, dermatan sulphate and chondroitin 4-sulphate interact at higher dilutions ('higher titres') with collagen than do chondroitin 6-sulphate or keratan sulphate. The results are relevant to studies of anti-collagen antibodies in connective tissue fluids and extracts, as well as to connective tissue physiology.


Epidermal chalone--past to present concept. How the constantly renewing epidermis maintains its homeostatic state remains a mystery. A "negative feedback" mechanism of control with chalone, a tissue-specific, species-nonspecific inhibitor of mitosis, has been suggested by Bullough and Laurence. Progress in isolating, purifying, and characterizing epidermal chalone has been slow and difficult because of the lack of a fast, assurate, and meaningful assay system. A G2 assay (depending upon mitotic counts) was used to obtain the data which served as the basis for postulating the existence of chalone. Since a control point with the G1 phase of the cell cycle is considered to be the one with physiologic implications, in vivo and in vitro assays with pure primary epidermal cell cultures have been used to investigate a G1 block. The incorporation of tritiated thymidine into DNA is often used as a measure of the rate of prokiferation. However, inhibition of thymidine transport and phosphorylation have been encountered in several systems and negate this method as an accurate measure of cell proliferation.


Steroid entry into rete testis fluid and the blood-testis barrier. Evidence that steroids enter rete testis fluid (RTF) from the blood at varying rates was obtained during i.v. infusions into rats. Testosterone and dehydroepiandrosterone were readily transferred into the fluid, whereas cholesterol was excluded. Between these extremes, the appearance of radioactivity in the RTF suggested the following order of entry rate: progesterone greater than pregnenolone greater than 5-alpha-reduced androgens greater than oestrogens greater than corticosteroids. Preliminary identification of metabolites in RTF and blood suggested that testosterone and dehydroepiandrosterone were transferred largely unchanged. Androstenedione and progesterone, however, were largely metabolized during transfer into the RTF, the former being transformed to testosterone. The results are used to discuss the nature of the blood-testis barrier to steroids and the source of androgens in the RTF.


The production of cardiac hypertrophy by tri-iodothyroacetic acid. Severe cardiac hypertrophy has been produced experimentally in rats by long-term, low-dose treatment with tri-iodothyroacetic acid. The dose used was insufficient to cause any apparent systemic or metabolic effect. It is suggested that similar iodinated substances in the blood in man, resulting from normal or abnormal thyroid hormone catabolism, may be causally related to some forms of cardiomyopathy.


Steroidogenesis in cerebral metastatic chorionepithelioma tissue in vitro. Homogenates of cerebral metastatic chorionepithelioma tissue were incubated with labelled dehydroepiandrosterone, pregnenolone or 20alpha-hydroxypregn-4-en-3-one. The metabolites of dehydroepiandrosterone which were isolated and identified were androstenedione, testosterone, oestrone, and oestradiol; no oestriol was detected. The only metabolite of pregnenolone and 20alpha-hydroxypregn-4-en-3-one isolated and identified was progesterone. No conversion of C-21 to C-19 steroids occurred in the metastatic chorionepithelioma tissue.


The mechanical properties of the abdominal cuticle of Rhodnius larvae. 1. The mechanical properties of loops of cuticle cut from the abdomens of 5th instar Rhodnius have been investigated. The cuticle shows pronounced viscoelastic behaviour. 2. Stress-relaxation tests show a continuously falling modulus over a wide range of times after the imposition of a strain. 3. Plasticized samples of cuticle show stress-relaxation curves which are shifted along the time axis towards earlier times by up to times 10-3. The modulus at any particular time after the imposition of strain is about 10 times lower than that of the unplasticized cuticle. 4. It is concluded that the mechanical properties of this cuticle are determined, at least for maintained stresses, largely by the matrix material. Chitin microfibrils may act as a reinforcing filler for short-term, rapid stresses. The cuticular macromolecules are probably not extensively cross-linked by primary bonds, though secondary interactions between them are probably important in the viscoelastic properties of the cuticle. 5. Plasticization probably involves a change in either the number or the strength of secondary interactions between the cuticle macromolecules, or both.


Proliferation in vivo of T lymphocytes reactive to histocompatibility alloantigens: a correction. Rat lymphocytes in mixed cultures can reutilize tritiated thymidine from labeled granulocytes. Shortly after thymidine injections in vivo, major effects on the frequency of labeled lymphocyte mitoses in peripheral blood cultures are introduced by 10-20% polymorph contamination, even though transfer of label via supernates is not demonstrable. Cold thymidine in the cultures prevents reutilization, and has permitted reevaluation of several previous conclusions concerning the life history of lymphocytes reactive to major histocompatibility alloantigens (HARC). Rather than being predominantly recently divided cells, HARC do not appear to have an age distribution, in blood or lymph, significantly different from the general recirculating lymphocyte population. However, the ability of immunization across strong allogeneic differences to increase markedly the proportion of young HARC among the specifically responsive population has been confirmed.


Effects of corticosteroid treatment and inflammation on the cellular content of blood and exudate in mice. Inflammatory exudates have been produced in mice by intraperitoneal injection of thioglycollate broth 24 hr and again 3 hr before collecting of the exudate. The first injection of broth exerts a "priming" effect leading to an enhanced response to the second injection. By this procedure more than 30 x 106 leucocytes of which 78 per cent. were polymorphs were obtained from each mouse. Administration of paramethasone reduced the number of cells in the exudate when given at the same time as broth but not when given 3 hr before collection of the exudate. In contrast paramethasone was equally effective when given either 24 hr or 3 hr before harvest in suppressing the appearance of intravenously injected pontamine sky blue in the exudate. It was striking that treatment with paramethasone which had reduced the number of polymorphs in the exudate had actually increased the number in blood.


Comparison of the effect of various antisera and cobra venom factor on inflammatory reactions in guinea-pig skin. II. The Arthus reaction and the local Shwartzman reaction. The ability of antisera to guinea-pig C3 to inhibit the Arthus and local Shwartzman reactions was studied. They were found to reduce the non-haemorrhagic component of the active and reversed passive Arthus reactions and to delay the appearance of the haemorrhage in the active Arthus reaction. Cobra venom factor, however, had no effect on the non-haemorrhagic components of these reactions and only delayed the appearance of the haemorrhage of the active Arthus reaction. There appeared to be a correlation between the serum complement level and the time taken for the haemorrhage to appear, and between the circulating platelet count and the extent of the non-haemorrhagic, oedematous component of the reaction. The haemorrhagic component of the local Shwartzman reaction was not affected by decomplementation with cobra venom factor. The ability of the antisera to inhibit the haemorrhage of the Shwartzman reaction was not dependent on lowering the serum complement titre. However, the haemorrhage was inhibited if the circulating platelet count was also reduced to very low numbers. Antiserum to zymosan alone had the same effect as anti-beta1C/beta1A globulin (zymosan) in blocking the reaction, although it did not alter the complement levels or the platelet counts. The possibility of an immunological cross-reactivity between zymosan and endotoxin in this action is discussed.


A histochemical study of the liver lesion induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) in rats. A histochemical study of plasma-membrane associated enzymes in rat liver demonstrated a significant lesion 3 days after a single oral dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin). The complete loss of canalicular ATPase reaction in the parenchymal cells of the centrilobular zone remained the prominent feature of the liver throughout the 6-wk period studied. Involvement of the periportal and midzonal regions occurred in moribund animals and improvement in the health of two surviving animals at 9 mth was associated with a normal distribution of ATPase in the liver. Qualitative changes in 5-nucleotidase and acid phosphatase were secondary to the parenchymal cell damage. This lesion supports the morphological evidence, reported previously, that the parenchymal cell plasma-membrane is a specific subcellular site of the toxic action of dioxin.


Biochemical characteristics of "young" and "old" erythrocytes of the newborn infant. Cord blood samples from ten term infants were fractionated into populations of young and old erythrocytes and compared with cells prepared in a similar fashion from eight normal adults. The old cell fraction from the newborn infants had very low phosphofructokinase activity and did not display the usual decline of activity of the enzymes phosphoglycerate kinase and enolase. In addition, the old cells from the newborn infants demonstrated impaired glucose consumption, which, upon analysis of the pattern of glycolytic intermediates, appeared to be a result of the phosphofructokinase deficiency. These findings suggest that cells produced earlier in gestation possess the developmental characteristics of fetal blood to a more significant degree and that these biochemical alterations may produce functional impairment.


Associated coarctation of the aorta and mitral valve disease: nine cases with surgical correction of both lesions in three. Nine cases of the combination of coarctation of the aorta and mitral stenosis were evaluated over a seven-year period. Symptoms did not usually cause distress in infancy, but began subtly with pneumonia or cardiac failure at about 2 years of age. Important clues were differences in blood pressure between the arms and legs, paroxysmal dyspnea, congestive heart failure, right ventricular hypertrophy, and left atrial enlargement. Cardiac catheterization studies showed elevated right ventricular and main pulmonary artery wedge pressures. These features in patients with coarctation of the aorta should suggest associated mitral valve disease. The importance of demonstrating associated valvular lesions, particularly mitral stenosis, is emphasized. Two of our children had successful repair of the coarctation of the aorta and mitral stenosis simultaneously. In a third child, resection of the coarctation was followed in six years by mitral valve replacement.


Cardiotonic steroids II: 3-deoxycardenolides and 3-deoxycardanolides. A series of compounds related to 3-deoxydigitoxigenin was prepared and assayed for inhibition of myocardial Na+,K+- adenosine triphosphatase. Although the relatively high activity of 3-deoxydigitoxin was confirmed, the corresponding 3beta,4beta-epoxide and a mixture of 2,3-olefins and 3,4-olefins were less active. 3-Deoxy compounds with variations at the 14-position and the butenolide ring were much less active than the corresponding 3beta-hydroxy analogs. Thus the activity of 3-deoxydigitoxigenin appears to be particularly susceptible to structural changes elsewhere in the molecule.


Mechanism of action of colchicine. I. Effect of colchicine and its analogs on the reversed passive Arthus reaction and the carrageenan-induced hindpaw edema in the rat. Colchicine and N-desacetyl-N-methylcolchicine suppressed both the reversed passive Arthus reaction and the carrageenan-induced edema in the rat. Colchicine, 2-desmethyl-colchicine glucoside and trimethylcolchicine acid had no effect on either model of inflammation. The ability or inability of these compounds to suppress the development of experimental inflammation correlated with their antimitotic activities. The findings lend support to the hypothesis that the anti-inflammatory and the antimitotic effects of colchicine may depend on the same basic, biophysical mechanism of action, i.e., the disruption of the microtubules.


Studies on the mechanism of inhibition of the red cell metabolism by cardiac glycosides. The purpose of this work was to test the previously suggested hypothesis that the inhibitory effect of ouabain on lactate production in human red cells is due to an interaction between phosphoglycerate kinase and (Na+ + k+)-activated adenosine triphosphatase (Na+,K+ATPase). An antibody to red cell phosphoglycerate kingase caused complete inhibition of the purified enzyme, whereas a portion of the phophoglycerate kinase activity of the red cell membranes was resistant to the antibody. When increasing amounts of the purified enzyme were added to the membranes, the antibody-resistant portion of the activity increased. The effects of the antibody and ouabain on lactate production from fructose-6,6-diphosphate in red cell hemolysates were studied. Ouabain, at a maximally effective concentration, produced about 30% inhibition of lactate formation. This value was doubled in the presence of the antibody. Red cell membranes, and rat brain Na+,K+-ATPase, did not catalyze the hydrolysis of 1,3-diphosphoglycerate. Ouabain did not affect the reactions of the Rapport-Luebering pathway of the red cells. These findings provide further support for the view that in red cells a membrane pool of phosphoglycerate kinase is oriented in the vicinity of Na+,K+-ATPase in a way that the product of each enzyme may be used as the immediate substrate of the other and that ouabain inhibits glycolysis by removing the regulatory effect of Na+,K+-ATPase on that portion of glycolysis which is channeled through this pool of phosphoglycerate kinase.


Skeleto-fusimotor axons in the hind-limb muscles of the cat. 1. Motor axons supplying various hind-limb muscles of the cat (flexor hallucis lingus, peroneus brevis, peroneus digiti quinti, tibialis anterior, soleus and tenuissimus) were identified as skeleto-fusimotor or beta axons because their repetitive stimulation elicited both the contraction of extrafusal muscle fibres and an increase in the rate of discharge of spindle primary endings which perisited after selective blockade of extrafusal neuromuscular junctions. 2. The conduction velocity of these axons ranged from 39 to 92 m/sec. 3. Of seventy-six beta axons, seventy-two had a dynamic action on the sensitivity to velocity of stretching of primary endings, four had a static action. 4. The dynamic action of six beta axons was observed only after the contraction of extrafusal muscle fibres was selectively suppressed. 5. Tendon organs can be activated by beta motor units.


Rate of flow of digesta and electrical activity of the small intestine in dogs and sheep. 1. Spiking activity of the small intestine in the conscious dog and sheep was recorded continuously from electrodes chronically implanted on the jejunum and summed at intervals of 20 sec. The activity was related to the transit time and flow rate of intestinal contents as estimated by phenol red and by dilution of continuous marker infusions respectively. Also in some sheep the flow of digesta was measured directly from a cannula in the proximal part of the jejunum, and also by use of an electromagnetic flow meter. 2. In the fasted dog and in sheep on a normal diet the intestinal activity was characterized by a migrating myo-electric complex comprising an irregular phase followed by a regular phase. These migrating myo-electric complexes occurred regularly after a period of inactivity at a frequency of 15-20/24 hr. In dogs after feeding, a continuous spiking activity appeared and persisted for periods of 7-8 hr. This was associated with much higher rates of flow and shorter transit times than were observed during fasting. In sheep, continuous spiking activity could be induced by intravenous injection of 5-hydroxytryptophan and this, similarly, was accompanied by a more rapid flow and a shorter transit time than recorded during the control period. 3. In both species the longest transit time occurred when a phenol red bolus was injected during the period of electrical inactivity. Relatively short transit times were observed when the bolus was administered just before the period of regular spiking activity. 4. When relaxation of the bowel was induced by intraperitoneal injection of hypertonic saline there was no spiking activity and the transit time for the infused solution was greatly lengthened, especially in the sheep. A noticeable flow of digestive contents persisted in the dog. 5. In the sheep the intestinal contents flowed intermittently during periods of 10-15 min and at the same frequency as the migrating myo-electric complex. Two thirds of this flow took place in the 4-6 min immediately preceding the periods of irregular spiking activity. 6. It is concluded that in the fasted dog and in the sheep the migrating myo-electric complex controls the pressure gradients on which the flow of intestinal contents depends. This is accomplished in the main by the prolonged phase of irregular spiking activity, and it is suggested that the regular spiking activity which follows it, though not in itself propulsive, serves as a barrier to prevent backflow of digesta into the quiescent part of the intestine. When continuous spiking activity is induced, by feeding in the dog and by injection of 5-hydroxytryptophan in the sheep, no part of the intestine is quiescent and the transit time is shortened by the incessant irregular spiking activity.


Permeability and porosity of dental casting investments. Permeability of various gypsum- and phosphate-bonded investments was measured during conventional burn-out procedures. Porosity determinations were made on specimens cooled to room temperature after burn-out. As a group, the gypsum-bonded investments were found to be more permeable than the phosphate-bonded investments. Two phosphate-bonded investments were determined to be relatively impermeable to gas flow, while another exhibited permeability comparable to that of the gypsum-bonded investments. In spite of differences in permeability, the porosity of each type of investment was nearly constant. The porosity of the phosphate-bonded investment was approximately three-fourths that of the gypsum investments. These investments were modified by the addition of varying amounts of acrylic polymer for the purpose of altering permeability. The addition of acrylic polymer increased porosity and permeability of all of the materials included in this investigation. The acrylic additives, however, had no effect on the permeability of relatively impermeable investments unless used in high concentration. The results of this investigation would tend to substantiate the need for special spruing and venting procedures.


[Disturbances of pulmonary ventilation in association with cardiac disease in infants (author's transl)]. Numerous causes exist for disturbances in pulmonary ventilation in the newborn and infant. Amongst the exobronchic causes, those of cardiovascular origin are not rare. The difficulty in radiological diagnosis varies in relation with the anatomo-radiological appearance of the lesions and their aetiology. Ventilation disturbances are not always obvious. Simple techniques (high penetration films, oblique views, films in inspiration and expiration) make possible the topographical localisation of the lesions and the detection of any obstructive factor. These films should also be adequate as far as the aetiological diagnosis is concerned: severe left-right shunt, pulmonary valve agenesis, fibro-elastosis, retrotracheal left pulmonary artery. Obviously, angiocardiography is often necessary to confirm and accurately determine the nature of the "cardiovascular" lesion responsible. Such disturbances of pulmonary ventilation may either reveal or complicate a cardiac or vascular disorder in the newborn or infant. They often present marked therapeutic problems.


The electrical dynamics of laparoscopic sterilization. The minimum voltage needed to perform laparoscopic sterilization was studied to learn more about prevention of bowel burns. Using a "CUT" waveform at very low levels, it is practical to coagulate fallopian tubes with as little as 100 volts peak. If electrosurgical cutting of fulguration (charring) is desired, the nimimum voltage is about 700 volts peak. The changes in electrical resistance of tissue were measured during coagulation of sample tissues and actual sterilizations. A discussion of basic electrosurgical principles is included.


A preliminary and intermediate report on a new laparoscopic tubal ring procedure. A preliminary report is given of a new laparoscopic sterilization technique utilizing silicone rubber rings. A description of the laparoscopic silicone ring applicator and its operative utilization is provided. This technique eliminates the need for thermoenergy to insure tubal destruction and thus removes the cause of intestinal burns. The potential for successful future recannulization procedures should be greater with this procedure. This operation can be performed with a single or double puncture technique; in addition, it can be performed with a suprapubic or vaginal approach. Three hundred one cases have been completed without incurring major problems. No statement can be made at this time about the incidence of failure.


Unusual uses of laparoscopy.

Unusual uses of laparoscopy. The marked enthusiasum recently seen in gynecologic endoscopy has brought forth numerous anecdotes of ingenuity. Without ingenuity, medicine and surgical skills would advance slowly. This anecdotal report is presented not only as information but as a stimulus to the endoscopist to consider new approaches to today's surgical problems. The laparoscope has been used to: 1. remove ectopic pregnancies, excise paraovarian cysts and retrieve foreign objects from the abdomen. 2. explore the abdomen for suspected trauma, 3. evaluate the abdomen and pleural cavity for metastatic disease, and 4. perform lumbar sympathectomy. These and other unusual uses of the laparoscope are presented for the "spelunkers" of modern-day endoscopy.


Antibodies to nucleic acids in human and murine lupus. Antibodies to nucleic acid are present in the sera of patients with systemic lupus erythematosus and in the sera of NZB/NZW F1 mice, an animal model for human systemic lupus erythematosus. Sera can be fractionated by sucrose-density gradient ultra-centrifugation and individual fractions assayed for binding of radioactive DNA, RNA or DNA: RNA hydridsive DNA, RNA or DNA: RNA hybrids. Immunoglobulin determinations can also be performed on these fractions. Antinucleic acid antibody activity can be recovered both in the 19S IgM and 7S IgG fractions. Individual variations from patient to patient suggest that more detailed analysis of immunoglobulin classes may provide insights into the pathogenesis of lupus.


Studies on the nature of circulating DNA in systemic lupus erythematosus (SLE). The isolation and purification of DNA from the sera of patients with SLE is described. Buoyant density determinations of this DNA using CsCl gradients suggest in some cases that it may be of endogenous origin. DNA from the synovial fluid of an SLE patient with septic arthritis was also shown to have a buoyant density of human DNA as well as being of considerable size variation. In addition, this synovial fluid contained antibody to DNA and evidence of DNA:anti-DNA complexes, suggesting that immune complexes composed of human DNA and antibody may be involved in the pathogenesis of the arthritis. Also described are preliminary experiments characterizing the conformation of the DNA present in DNA:anti-DNA immune complexes.


Intraspinal epidermoid tumor of the sacral canal presenting with urinary retention. A case of epidermoid tumor of the sacral area with S3 root irritation resulting in bladder dysfunction is presented and its possible relationship to spinal puncture is discussed. Precautions should be taken to prevent this complication by using a stylet instead of a plain needle. The recognition of this complication should always be in the differential diagnosis of a child with back pain, gait abnormality and bladder dysfunction with a history of spinal puncture.


125-I-labeled fibrinogen scanning. Use in the diagnosis of venous thrombosis. Venous thrombosis is often asymptomatic in patients in whom major pulmonary embolism develops. When used expectantly, iodine 125-labeled fibrinogen scanning is a very sensitive method for detecting subclinical leg vein thrombi. Fibrinogen scanning is less useful for the diagnosis of established venous thrombosis, but is valuable for detecting extension of venographically diagnosed calf vein thrombosis. The technique is safe if fibrinogen is obtained from carefully screened donors. The limitations of the method include its inability to distinguish between superficial and deep venous thrombi, and its sensitivity to fibrin in hematoma and inflammatory exudates. Though the results agree closely with those of phlebography, scanning seems less reliable for detecting femoral vein than calf vein thrombi and is insensitive to thrombi above the inguinal ligament. Screening for these major thrombi may be improved by combining fibrinogen scanning with impedance plethysmography or ultrasonic examination.


Echocardiographic classification of hypertensive heart disease. A correlative study with clinical features. Forty patients with systemic hypertension were classified into 4 types based on the left ventricular echocardiographic findings. Patients with normal left ventricular echogram, type I, showed little clinical symptoms and signs of hypertensive involvement. Higher systolic pressure and marked hypertensive retinal and renal changes were observed in patients with symmetric hypertrophy of the left ventricle, type II. Congestive heart failure was dominantly present in those with dilatation of the left ventricle, type IV. High voltages and marked ST-T changes in electrocardiogram were usually found in patients with asymmetric septal hypertrophy, type III, while retinal and renal damages were mild. Left ventriculograms obtained from 6 cases in type III also revealed hypertrophy of the interventricular septum and one of them demonstrated left ventricular outflow tract obstruction. These cardiac features in type III, which are quite similar to those in hypertrophic cardiomyopathy, seemed to be a secondary change induced by systemic hypertension on the basis of some predisposition.


Effects of ruthenium red, quinacrine hydrochloride, ethacrynic acid and 2,4-dinitrophenol on the water receptor of the frog tongue. Effects of some chemicals, which are known as inhibitors of Ca2+-dependent ATPases, on the water receptor of the frog tongue were examined by using single fungiform papilla preparations. When a sufficient amount of ruthenium red, quinacrine hydrochloride, ethacrynic acid or 2,4-dinitrophenol was added to the standard stimulating solution (5mM CaCl2+100 mM NaCl), which has been shown to stimulate sufficiently the water receptor of the frog tongue, no neural response was elicited. The concentrations necessary for 50% inhibition were approximately 3 X 10(-6)M for ruthenium red, 1 X 10(-5) M for quinacrine hydrochloride, 1 X 10 (-3) M for ethacrynic acid and 2 X 10(-4) M for 2,4-dinitrophenol. Organic mercurials, mersalyl acid and p-chloromercuribenzoic acid, had no effect on the nueral response, but repeated application of these chemicals led to a permanent depression in receptor activity. Ouabain had no effect on either the neural response or receptor activity. These observations indicate that the receptor molecule of the frog water receptor has a similar property to that of the Ca2+-dependent ATPase of red-cell membrane in respect to the susceptibility to inhibitors.


[Clinical picture and variants in the course of chronic postinfarction aneurysms of the heart]. The study is based on the analysis of 268 patients with chronic postinfarction aneurysms of the heart, 190 of whom were operated. Clinical characteristics of postinfarction cardiac aneurysms are presented, and several symptoms typical for this pathology are singled out: pericardial pulsation, low pulse volume, tachycardia, outward shift of the left margin of the heart, galloping atrial rhythm. Proceeding from a careful analysis of the clinical material an original classification of the variants of the clinical course of chronic postinfarction carrdiac aneurysms is introduced: 1) with prevailing chronic coronary insufficiency; 2) with combined chronic coronary and cardiac insufficiency; 3) with prevailing cardiac insufficiency.