Capsulectomy of the metacarpophalangeal and proximal interphalangeal joints. From July 1975, through March, 1978, 105 metacarpophalangeal joint capsulectomies in 37 patients, 47 dorsal proximal interphalangeal (PIP) capsulectomies in 26 patients, and 65 volar PIP capsulectomies in 41 patients were performed. The procedures were done after conservative methods had failed, and the results were tabulated in each area by diagnostic category. In the major group in which stiffness resulted from fracture and crushing injuries, average gains of 13 degrees to 18 degrees of active motion were achieved, with a change in arc of motion, while nerve paralysis patients did substantially better. The study should clarify expectations from the procedure; functional gains may still be significant.


Congenital anomaly of the thumb: absent flexor pollicis longus tendon. A 6-year-old girl with congenital loss of flexion in the right thumb was found to have no flexor pollicis longus tendon. All other median-nerve-innervated structures were present and normal.


Simultaneous dislocation of the interphalangeal and metacarpophalangeal joints of the thumb: a case report. A complete dislocation of the proximal phalanx of the thumb is presented. At 6 months after the injury, the patient had a painless right thumb with a full range of motion. No such similar case has been found in the literature.


Some consequences of the new UK 'recommended daily amounts of food energy and nutrients' for evaluating food consumption surveys. The amounts of energy and nutrients which have been recommended by the Department of Health and Social Security in 1969 and 1979 are compared. For the nation as a whole, the recommendations for energy and protein have decreased by 6 per cent; thus, the adequacy of an otherwise unchanged diet will appear 6 per cent higher than before. The changes for iron, calcium and the three main B-vitamins are smaller, while vitamin C and vitamin A are virtually unaffected. Nevertheless, as the method of calculating retinol equivalents in the diet should now conform to worldwide practice, the amount of vitamin A in the average diet will appear to be about 6 per cent less than before. It will be no longer practical to evaluate dietary intakes of vitamin D, but there are now official recommendations for folic acid.


Rationalisation of therapeutic diets in the Netherlands. In 1976 a project was started by The Netherlands Education Bureau for Food and Nutrition to rationalise therapeutic diets and reorganise hospital catering departments. The first part of the work--rationalisation of therapeutic diets, including the introduction of a 'national dietetic instruction form'--is reported here.


Assessment of nutritional status: the biochemical approach and its problems in liver disease. The role of laboratory tests in the assessment of nutritional status is reviewed. Methods commonly used in evaluating adequacy of energy, protein, mineral and vitamin intake are discussed, and a protocol for nutritional assessment and monitoring is presented. The specific nutritional problems found in liver disease, and the effects these have on this protocol are considered.


Alcohol and the liver.

Alcohol and the liver. This paper discusses briefly the definition of alcoholism, the extent of the problem in Britain, the roles of ethanol toxicity and malnutrition in causing liver damage, and the types of liver damage seen in alcoholics.


Has cimetidine revolutionised the medical treatment of peptic ulceration? Cimetidine inhibits gastric-acid secretion, whatever the stimulant. It is superior to placebo for the healing of both duodenal and gastric ulceration, but it does not cure peptic ulceration. When the drug is stopped ulcers often return. Maintenance treatment with cimetidine taken only at bedtime eliminates duodenal ulcer recurrence in most patients.


Getting the message across.

Getting the message across. Messages do not simple 'get across'. The author points out the importance (1) of seeing good communication as an effort of understanding between human beings and (2) of dismissing any idea of a single communicator's skill. There are many and different skills to be applied.


Look after yourself: monitoring the effects of a campaign. A long-term campaign for better health was launched by the Health Education Council in the UK in January 1978. Preliminary findings from surveys evaluating the campaign are presented.


Role of the nurse specialist in stoma care. The work of the stoma-care nurse is discussed. Emphasis is laid on the need to liaise with other health professionals also concerned with the patient in hospital and in the community. Some specific problems of stoma patients are also discussed.


Modulation of platelet activation by native DNA. Native DNA (dsDNA) was found to induce the aggregation of isolated human platelets and the release of platelet 5HT; this activation was inhibited by both theophylline and TYA, suggesting a role for cAMP and metabolic products formed from arachidonate. By contrast, nonaggregating amounts of dsDNA inhibited platelet activation induced by collagen or thrombin. This inhibition, which could be overcome by use of greater amounts of the stimulatory agents, was not associated with the loss of platelet viability. Activation of platelets by dsDNA was not observed in plasma or in isolated platelet systems to which small amounts of cell-free plasma were added. However, dsDNA maintained in plasma its ability to inhibit platelet aggregation induced by collagen and thrombin. RNA and single-stranded DNA failed to induce platelet aggregation or release of 5HT and to block the platelet activation stimulated by dsDNA. Further, dsDNA did not significantly inhibit platelet aggregation in platelet-rich plasma stimulated by ADP or epinephrine. These data implicate dsDNA as a selective and potentially important activator and modulator of platelet responsiveness.


Biosynthesis and post-synthetic modification of a precursor (pro-C5) of the fifth component of mouse complement (C5). Mouse peritoneal macrophages synthesized and secreted a precursor (pro-C5) of the fifth component of serum complement (C5) in short-term tissue culture. Approximately 0.2% of the newly synthesized intracellular protein and 0.8% of secreted protein were precipitable with antiserum to mouse C5. The precursor is similar in size to the native serum protein (210,000 daltons), but consists of a single polypeptide chain. In contrast, serum C5 consists of two polypeptide chains (m.w. 125,000 and 83,000) linked by disulfide bridges. An electrophoretic variant of pro-C5 distinct from intracellular C5 was detected in medium from macrophage cultures.


Prevention of transplantable tumors by adoptive transfer of spleen cells from immunized rats. Fischer 344 rats were specifically hyperimmunized with allogeneic, nonvirus-producing [Kirsten murine sarcoma virus (KiMSV)] or syngeneic, virus-producing [KiMSV (Rasheed)] rat tumors. Spleen cells taken from these rats adoptively transferred protection against a 100 to 1,000 X rat tumor dose50 cell challenge with several different transplantable rat tumors. Protection was obtained with spleen cells after removal of adherent cells and macrophages but not peritoneal cells. The spleen cells were not directly cytotoxic but required more than 3 days residence in the recipient before protecting the recipient against challenge. No protection against tumor cell challenge was observed when spleen cells were lethally x-ray irradiated before injection into nontreated rats. Spleen cells taken from rats immunized with normal histocompatibility antigens did not protect in this test system.


Generation of biologic activity from the purified alpha-chain of C5. The alpha- and beta-chains of the fifth component of human complement (C5) have been isolated and their physical and biologic properties have been characterized. After electrophoresis in SDS-poly-acrylamide slab gels, the chains were eluted from the gels and extensively dialyzed. The amino acid composition of each chain was then determined. On SDS-gels, the alpha-chain was PAS positive whereas the beta-chain revealed little or no staining. As a measure of chemotactic activity, the lysosomal enzyme-releasing activity of both chains was examined before and after trypsinization; activity resided almost entirely in the alpha subunit. Reconstitution of C5 from its isolated chains, followed by trypsinization resulted in slightly less activity than that from the individual alpha subunit. These findings provide direct support for previous evidence suggesting the biologic activity for neutrophils derives entirely from the alpha subunit of C5. This represents the first report of direct recovery of biologic activity from a purified subunit chain of C5.


Structural studies on the murine Ia alloantigens. VI. Evidence that both subunits of the I-A alloantigen are encoded by the I-A subregion. The alpha and beta subunits of the murine I-A alloantigens from several H-2 haplotypes were examined by comparative tryptic peptide mapping by using double label (3H and 14C) techniques. Significant structural variation between alleles was detected in both subunits. Tryptic digests of the alpha polypeptides from s, b, and d showed only 65% co-elution with k; beta-chains from s, b, d, and r were about 50% similar to the k beta subunit. Peptide analysis of the Ak subunits from intra-H-2 recombinant strains indicated that both the alpha and beta polypeptides are encoded within the I-A subregion.


Plasminogen activator production by human monocytes. I. Enhancement by activated lymphocytes and lymphocyte products. Human monocytes, but not nylon wool column-nonadherent lymphocytes, produce plasminogen activator. The activity is found only in association with intact cells. Exposure of monocytes to activated lymphocytes or to lymphokine-rich supernatants enhances monocyte plasminogen activator production. The assay allows assessment of baseline and activated human monocyte function.


Autoimmune reactions against liver cells by syngeneic neuraminidase-treated lymphocytes. We have found that the entrapment of neuraminidase-treated lymphocytes in the liver leads to the induction of autoimmune cellular cytotoxic reactions. Lymphocytes from mouse spleen and thymus were incubated with neuraminidase in vitro and injected i.v. into syngeneic recipients. Lymphocytic infiltrations into the liver were seen 7 days later with both types of cells. After repeated weekly injections of asialo-lymphocytes, destruction of liver tissue became apparent. Electron-microscopic studies showed that hepatocytes, fat storage cells, and endothelial cells were affected, mainly at the hepatic periphery. It is concluded that the adhesion of asialo-lymphocytes to liver cells induces their cytotoxic activity. Similar reactions may occur after paramyxovirus infection due to the action of viral neuraminidase.


Immunopotentiating effects of amphotericin B. I. Enhanced contact sensitivity in mice. Contact sensitivity responses to dinitrofluorobenzene (DNFB) or oxazolone were enhanced by amphotericin B (AmB) administration. This adjuvant effect of AmB was documented in mice by ear thickness measurements, ear histology, and the 5-iodo-2'-deoxyuridine-125I ear assay. The optimum immunopotentiating effect of AmB required its simultaneous administration at the time of skin sensitization. AmB-induced adjuvant effects were also observed in adoptive transfer experiments in which syngeneic recipients of lymph node cells from animals sensitized with DNFB plus AmB gave stronger contact sensitivity responses than recipients of cells from mice sensitized with DNFB alone. AmB also interfered with tolerance induction by i.v. dinitrobenzene sulfonic acid, suggesting that its adjuvant effects involve inhibition of suppressor cells or their precursors.


Immunopotentiating effects of amphotericin B. II. Enhanced in vitro proliferative responses of murine lymphocytes. Enhanced in vitro proliferative responses to DNBSO3 were seen in lymph node cells and spleen cells after in vivo sensitization of mice with DNFB plus AmB compared with mice primed with DNFB alone. The T cell proliferation in the nylon column nonadherent fraction for both groups was highly similar, and the enhanced lymph node cell proliferation with AmB was demonstrated to be in the nylon adherent population consisting of both T and B cells. These and earlier studies of immunopotentiation by AmB are consistent with a mechanism that depends on selective interaction of the polyene with a subset of T cells and a resultant impairment of the normally induced suppressor regulation that limits the magnitude and duration of immune responses.


Inhibition of human lymphocyte E-rosette formation by oxygenated sterols. 25-Hydroxycholesterol, 20 alpha-hydroxycholesterol, 7 alpha-hydroxycholesterol, and 5 alpha-hydroxy-6-ketocholestanol, when added to cultures of human lymphocytes in lipoprotein-depleted medium (LPDM) at a concentration of 2.5 x 10(-6) M, inhibit E-rosette formation with sheep red blood cells. 20 alpha-Hydroxycholesterol, 7 alpha-hydroxycholesterol, and 5 alpha-hydroxy-6-ketocholestanol are more potent inhibitors than 25-hydroxycholesterol. The inhibitory effect of 5 alpha-hydroxy-6-ketocholestanol on E-rosette formation appears after 15 min of exposure; with the other three compounds, an exposure time of 18 hr is necessary. The inhibitory effect of E-rosette formation can be abolished by addition of free cholesterol, low-density lipoprotein, or high-density lipoprotein to the LPDM or by incubation of the cells in normal AB serum, but not by the addition of mevalonic acid to the LPDM. These observations suggest that the capacity of oxygenated sterol compounds (OSC) to inhibit E-rosette formation is independent of their inhibitory effect on sterol synthesis. It is possible that OSC inhibit E-rosette formation as a consequence of their insertion into the lymphocyte membrane as cholesterol analogues.


Interaction between components of the human classical complement pathway and immobilized Cibacron Blue F3GA. The interaction between the complement components in human serum and the dye, Cibacron Blue F3GA, immobilized on cross-linked agarose (Affi-Gel Blue) has been studied. All nine components of the classical complement pathway bound to the dye and could be recovered using a linear salt gradient. With the exception of C5 and C8, all the components were eluted over a narrow NaCl concentration range, with the following yields: C1, 17%; C2, 69%; C3, 92%; C4, 87%; C6, 105%; C7, 109%; C9, 128%. C5 and C8 eluted throughout the NaCl gradient with yields of 103% and 14%, respectively. Since all components could be eluted without substantial contamination by albumin or IgG, this procedure may prove valuable as an initial step in the purification of complement components. In addition, the ability of immobilized Cibacron Blue F3GA to physicallly remove complement components may prove useful for both the decomplementation of serum and in elucidating the role of complement in immunological reactions.


The influence of antigen properties on the conditions required to elute antibodies from immunoadsorbents. Immunoadsorbents were used to purify a number of antibodies. Using pH 2.0 acid conditions alone it was possible to elute antibodies raised against human growth hormone (HGH) and Fc fragments from such immunoadsorbents with 50% or better recovery of antibody activity. However, to elute antibodies raised against triiodothyronine and cortisol required 6 M guanidine HCl, pH 2.0. The avidities of the purified antibodies were similar to those of the non-purified antibodies. The purified antibodies were stable in solution at 4 degrees C for at least six months.


A polyethylene glycol radioimmunoprecipitation assay for human immunoglobulin G. A polyethylene glycol (PEG) radioimmunoprecipitation assay for human IgG is described that is sufficiently sensitive to detect 0.5 ng of IgG. This model antibody-antigen system was also used to study the stoichiometries of PEG-precipitated complexes. Our results suggest that the presence of PEG may affect the stoichiometry of the complexes which precipitate from solution.


Cell separation with staphylococcal protein A immunoadsorbent surfaces. An 'indirect" cell separation technique was developed using protein A, in which each specific antiserum was first reacted with the cells to be processed. Separation was then achieved on antibody/protein A immunoadsorbent surfaces, where the final antibody coupled to the protein A was one which was directed against the specific antisera previously reacted with the cells. Only small volumes of specific antisera were required to effect separation either on a small or large scale with excellent specificity, cell viability and total cell recoveries. Theoretically, the technique would function with any Ig class of specific antisera from any species and would not be restricted to mammalian protein A binding IgG specific antisera, allowing considerable expansion in the number of antisera able to be utilized.


A simple method of evaluating delayed type hypersensitivity in mice. Delayed type hypersensitivity reactions in mice sensitized to dinitrofluorobenzene were assessed by measuring the increase in weight of the ears challenged with antigen. The method has proved reliable for assessment of DTH reactions when results are expressed as weight ratios between challenged and unchallenged ears.


Use of C6- and C7-deficient human sera in quantitative hemolytic assays for C6 and C7. Quantitative hemolytic assays for C6 and C7 using as R reagents sera from patients deficient in these components are described. The assays gave linear results. Normal range for serum C6 was found to be 21,400--41,700 C6 hemolytic units/ml; for serum C7 the normal range was 5540--9860 C7 hemolytic units/ml.


Metrizamide gradient purification of mouse tumor cells. A one-step isopycnic density gradient centrifugation procedure is described for purification of highly viable and homogeneous tumor cells from a variety of solid mouse tumors. Mechanically suspended cells are layered onto preformed continuous gradients of medium 199-buffered 7--33% metrizamide (density range 1.05--1.20 g/cu. cm) isoosmotic with mouse plasma and centrifuged for 30 min. Large numbers of tumor cells, generally 85--95% viable and free from 80--95% of contaminating host lymphoid and phagocytic cells and erythrocytes, were consistently recovered from fractionated thymomas, melanomas, and fibrosarcomas. By a variety of criteria, cell surface and other biological properties of gradient-purified tumor cells were normal.


Immunoglobulin allotypes and response to tetanus toxoid in Papua, New Guinea. The data indicates that Papuans with the Austronesian haplotype Gmf,a;b produce significantly higher levels of tetanus specific antibody, than Papuans without this haplotype, suggesting that response to tetanus toxoid in man is associated with the Gm phenotype of the individual. Km (1) status was not related to response.


Discrimination of Antibodies to Hepatitis B surface antigen from antibodies to inherited serum protein variants in immune sera of human and animal origin. Antisera to hepatitis B surface (HBs) antigen, both of human and rabbit origin, have been examined. The anti-HBs serum derived from a multiply transfused patient did, in addition, contain antibodies directed against the inherited beta-lipoprotein antigen Ag(x), whereas one of the rabbit immune sera also contained antibody to the inherited Lp(a) antigen. Thus, in human, as well as in animal anti-HBs sera, antibodies to inherited normal serum antigens may cause false positive reactions. This problem may be overcome by absorption procedures if appropriate control systems are available. False positive reactions caused by the Ag(x) antigen may also be avoided by use of agarose as supporting medium for the test, as Ag(x) precipitin lines do not appear in this medium. An undialysable high molecular weight component may be obtained from Oxoid "Ionagar" by washing. When this is added to agarose the Ag(x) antigen reaction appears also in this medium.


The specialized junctions between Merkel cell and neurite: an electron microscopic study. Longitudinal serial sections of one half of the entire sinus hair of a mouse were examined by the electron microscope. Three neurites entering the outer root sheath from the perifollicular blood sinus were encountered. These were separate nerve trunks from those connected with perifollicular tactile nerve endings and exclusively innervated intrafollicular Merkel cells. Two types of specialized junctions were observed at the contact regions between Merkel cell plasma membrane and neurite plasma membrane: (i) desmosome-like structures in which small clear vesicles and/or the large cored vesicles of neurite and thicker membrane (post-synaptic?) of apposed Merkel cell were found ant (ii) synapse-like structures in which Merkel cell granules were concentrated near the plasma membrane and the membrane of the apposed neurite was usually thicker (post-synaptic?). In some of the synapse-like junctions the limiting membrane of Merkel cell granules fused with the Merkel cell plasma membrane and its content seemed to be discharged into the intercellular space. This suggested actual exocytotic secretion of Merkel cell granules. Juxtaposition of 2 types of junctions, i.e. (i) and (ii) above, was also found. This suggested the possibility that the reciprocal synapse would be present between Merkel cells and afferent neurites.


The effect on rhino mouse skin of agents which influence keratinization and exfoliation. The skin of the rhino mouse, an allelic variant of the hariless mouse, contains deep dermal cysts and huge numbers of hornfilled utriculi which resemble comedones. Chemicals which influence either differentiation or desquamation of horny cells were applied topically twice daily for up to 6 weeks. Except for the dermal cysts, the gross epithelial abnormalities were almost completely corrected by retinoic acid in a dose-dependent fashion. Salicylic acid caused partial emptying of the horny masses, but the utriculi did not regress. Lactic acid, propylene glycol and benzoyl peroxide had minor effects on keratinization and exfoliation. The rhino mouse is a suitable model for assessing chemicals which affect epithelial differentiation (retinoids)or which promote loss of cohesion between horny cells (descaling agents).


Changes in carbohydrate expression of lichen planus affected oral epithelial cell membranes. The purpose of the present study was to examine possible epithelial cell membrane changes in lichen planus affected oral mucosa by the use of the lectins concanavalin-A and ricinus communis agglutinin fraction I. It was shown that these lectins in contrast to what was found in normal epithelium do not bind to the cell membranes of basal cells of lichen planus affected epithelium. The lack of binding of lectins suggests changes in exposed carbohydrates at the surfaces of the cells. The findings may support the hypothesis of antigenic changes in the lichen planus affected epithelium.


Specific binding of prostaglandin E2 to membrane preparations from human skin: receptor modulation by UVB-irradiation and chemical agents. Human skin membranes bind prostaglandin E2 (PGE2) with high affinity (with an apparent dissociation constant, Kd, of 3.14 X 10(-9) M) and specificity. This binding is inhibited by trypsin or heat treatment suggesting that PGE2 receptors have protein components. Exposure of the membranes to ultraviolet irradiation (UVB) resulted in the loss of the membrane binding capacity for PGE2. This UVB-ihibitory effect could be prevented by 5,5'-dithiobis-(2-nitrobenzoic acid), a known protein sulfhydryl-oxidizing agent and alpha-tocopherol, a known lipid anti-oxidant. These results suggest that UVB-irradiation possibly initiate the reduction of critical protein disulfide groups and the peroxication of lipids in the membranes, which are essential for the receptor-PGE2 interaction.


Excretion pathways of amphotericin B.

Excretion pathways of amphotericin B. The role of the biliary system in excretion of amphotericin B was explored in a dog model that allowed either external diversion of all bile or complete biliary obstruction. In dogs with biliary diversion, which were given a single dose of amphotericin B intravenously, excretion of amphotericin B in the bile lasted for seven to 10 days and accounted for only 3% +/- 2% (mean +/- SD) of the dose, whereas excretion in the urine was prolonged (23--35 days) and greater (21% +/- 5% of the dose); the stool contained no amphotericin B. However, bile salt depletion may have depressed biliary excretion of amphotericin B: in a dog with an intact biliary system, 19% of the dose was excreted in the stool over 11 days. In dogs given amphotericin B daily, serum levels were 19% +/- 3% higher during periods of biliary obstruction than during periods of free bile flow (P less than 0.05). Thus, excretion of amphotericin B in the bile (less than or equal to 19% of the dose) and in the urine (21% of the dose) accounted for a minority of total drug clearance. Nevertheless, prolonged excretion of amphotericin B by these routes after a single dose suggests that infrequent doses of amphotericin B may provide effective treatment for certain forms of fungal infection.


Furosemide enhancement of experimental gentamicin nephrotoxicity: comparison of functional and morphological changes with activities of urinary enzymes. Dogs were given gentamicin (10 mg/kg) intramuscularly every 8 hr for 10 days. Levels of serum creatinine rose by day 6 (0.91 +/- 0.08 vs. 0.75 +/- 0.02 mg/dl for controls, P less than 0.05) and of blood urea nitrogen by day 8 (24.3 +/- 4.80 vs. 16.1 +/- 0.90 mg/dl for controls, P less than 0.05). Gentamicin nephrotoxicity occurred earlier and was more marked when furosemide (2 mg/kg) was added: the level of serum creatinine by day 6 was 1.62 +/- 0.25 mg/dl (P less than 0.05), and the level of blood urea nitrogen by day 8 was 181 +/- 23.5 mg/dl (P less than 0.01). Elevations in the activities of the urinary enzymes beta-glucuronidase, N-acetyl-beta-glucosaminidase, and muramidase preceded rises in levels of serum creatinine and blood urea nitrogen. Examination of serial percutaneous renal biopsy specimens showed that gentamicin administration was associated with hyaline droplet degeneration, lysosomal changes, and, later, cell necrosis (primarily of the proximal tubules). Changes in renal morphology were more severe and occurred earlier when furosemide was administered concomitantly. In summary, furosemide enhanced gentamicin nephrotoxicity. Enzymuria was an early sign of gentamicin nephrotoxicity.


Microbial colonization and hepatic abnormalities in jejunoileal bypass with resection, ileal mucous fistula, and ileocolostomy. A role for bacterial colonization of the intestines in the pathogenesis of hepatic abnormalities after jejunoileal bypass was sought. Dogs were divided into groups according to the disposition of the bypassed segment; resection (group I), exteriorization as an ileal mucous fistula (group II), and drainage via an ileocolonic anastomosis (group III). Weight loss, abnormalities in liver function, and hepatic steatosis were significantly greater in groups II and III than in group I. Concomitantly, there was a significant increase in the total number of bacterial colony-forming units in groups II and III. Moreover, a greater number of specific anaerobic genera was isolated in group III than in group II. It is concluded that: (1) retention of the bypassed intestinal segment is associated with greater changes in liver function and structure than when the segment is resected; (2) the changes in the liver correlated with bacterial proliferation in the bypassed intestinal segment; and (3) despite a greater number of anaerobic genera in the ileocolostomy than in the mucous fistula, both procedures were associated with hepatic abnormalities.


Extracellular factors, blood group antigens, and bacteriophage of nephritogenic and nonnephritogenic strains of M-type 12 streptococci. Strains of M-type 12 streptococci from 18 patients with acute glomerulonephritis and 18 patients with uncomplicated pharyngitis were analyzed for in vitro production of streptolysin O, diphosphopyridine nucleotidase, hyaluronidase, streptokinase, streptolysin S, proteinase, hyaluronic acid, and fibrinogen-precipitating factor. In addition, relations to blood group antigens, lysogeny, and susceptibility to bacteriophage were determined. No significant differences were found between strains from nephritic and nonnephritic patients. By not indicating a role in the pathogenesis of poststreptococcal acute glomerulonephritis for any of the factors studied, these observations diminish the probability that these factors are of specific importance in this disease and thus direct our attention elsewhere.


Failure to detect hepatitis B surface antigen (HBsAg) in feces of HBsAg-positive persons. Since previous studies did not provide conclusive data regarding the presence of hepatitis B surface antigen (HBsAg) in the feces of HBsAg carriers, the feces of 20 HBsAg carriers and six patients with HBsAg-positive active liver disease were examined with use of a reproducible method for concentrating and detecting HBsAg in feces in which bovine serum, which has been shown to protect HBsAg destruction by fecal components, was added to the fecal specimens. HBsAg was not detected in 66 fecal specimens from the 26 HBs-Ag-positive persons. Furthermore, HBsAg could not be detected in the feces of a carrier who had ingested 4 ml of his own serum. This finding suggests that there are factors in the gastrointestinal tract that interfere with the immunoreactivity of the HBsAg and possibly destroy the hepatitis B virus. These findings explain why oral/fecal spread does not play a major role in the transmission of hepatitis B virus.


Persistence of hepatitis B surface antigen in the bedbug Cimex hemipterus (Fabr.). The fate of hepatitis B virus in the bedbug was investigated to assess this insect's potential as a vector. Colony-reared Cimex hemipterus (Fabr.) were fed once on blood positive for hepatitis B surface antigen (HBsAg). The insects were sampled at intervals thereafter and tested for HBsAg by radioimmunoassay. HBsAg persisted for up to six weeks in the bedbug's body after a single HBsAg-positive meal, during which time several further HBsAg-negative blood meals were taken. This result explains the high rates of field infection in bedbugs and further supports the hypothesis that bedbugs may play a role in transmission of hepatitis B virus.


[Experimental and clinical studies on the Jikei total knee prosthesis (author's transl)]. Total knee prostheses for a dog were developed and were inserted into knee joints of seven adult dogs to study the mobility of the artificial joint, the degree of fixation to bone and the durability of the prosthesis, to examine the reactions in the surrounding tissues and the internal organs in the JIKEI total knee prosthesis in clinical use. As a result of surgery, it was proved that the length in the hinged part of the prosthesis needed to be seventy percent of the length of the excised area of the joint to gain good mobility and supportability of the knee joint after surgery. As a result of findings based on necropsy, wear at the hinged part of the prosthesis was mild and debris caused in wear was not harmful for the living body if breakage does not occur there. Histopathological findings revealed that collagenous fibrous tissue with new bone formation was observed on the inner wall of the bone around the stem of the prosthesis four months after surgery. Then it was proved that the fixation of the prosthesis to the bone was sufficient. Therefore the author would like to mention that bone cement is not necessary for the fixation between bone and prostheses. The JIKEI total knee prosthesis (universal type) based on the results of these experimental studies has been developed and was used to the clinical cases. The clinical results until four years after surgery have been most satisfactory with loss of pain, good supportability and movement of joints in almost all of the cases.


[A study on experimental pyogenic osteomyelitis. 1. The preferential site of hematogenous osteomyelitis. 2. The role of foreign body in hematogenous infection (author's transl)]. The author undertook the experiments as described below in order to determine the preferential site of hematogenous osteomyelitis and possible effects of foreign bodies in the bone marrow on development of infection. I) In the first experiment, mice were inoculated with 10(7) cells of staphylococcus aureus intravenously into the tail vein and examined for the distribution and proliferation of the organisms in the bones, all over the body. It was found that the orgaisms in the blood stream were distributed to the bones all over the body almost evenly and that after prolonged observation, remarkable proliferation was noted in both femur and tibia. This bacteriological fact was supported by roentgenographic examination of all the bones. II) In the second experiment, inoculation of staphylococcus aureus into the tail vein was made after a foreign body (No. 5 sterile silk thread of 3 mm in lenght) had been inserted into the proximal metaphysis of a tibia of mice. The tibia of both legs was divided into 3 parts: proximal metaphysis, diaphysis and distal metaphysis. Observation were made for the distribution of organisms and state of proliferation in each of these three parts. In order to elucidate how the inserted foreign body promoted the establishment of infection, the group inserted with silk thread was compared with the control group (un-inserted group). The following results were obtained: 1. Although after 2 weeks of inoculation, the foreign body exerted no influences on the distribution and proliferation of organisms, but it was experimentally demonstrated to play a very important role for retention of organisms and maintenance of infection for a long period. 2. When the inoculum size was 10(6) to 10(7), the organisms were distributed evenly throughout the tibia. However, in the diaphysis the organisms tended to disappear, while in the metaphyseal area the organisms were easy to proliferate. It was also shown that at least 10(5) of organisms were needed for the establishment of infection and that the success or failure of establishment of infection is determined usually within 24 hours after inoculation of the organisms. As described above, from this study it was demonstrated that the femur and tibia were the preferential site of hematogenous osteomyelitis and that metaphyseal areas of the long bones were commonly affected. Once hematogenous invasion of organisms occurred, administration of antibiotics should be started as soon as possible, no later than 24 hours. According to our common experience, removal of foreign body is necessary for the complete cure of the injection.


Histochemical study of the human intervertebral disc. The human intervertebral discs which were obtained by cadavers and anterior discectomy are investigated histochemically. Chondroitin-4S, chondroitin-6S, dermatan sulfate, hyaluronic acid and keratan sulfate were detected in the human intervertebral disc by various histochemical methods. pH2.5, pH1.1 toluidin blue metachromasia and 0.4M MgCl2 alcianophilia became weaker with increasing age, and the herniated disc were weaker than controlled discs in the same age group. Chondroitin-4S and chondroitin-6S were distributed throughout the discs. There was no clear localization of the various glycosaminoglycans in the human intervertebral disc, with the exception of keratan sulfate. There was no histologically and histochemically observable difference in the cervical and lumbar discs.


Gait patterns of the diseased knee joint. Biomechanical gait analyses were carried out on 21 patients in whom 11 had bilateral and 10 unilateral disease of the knee joint. Similar studies, for use as controls were made on 7 normal volunteers. The data, on which the analyses were based, were obtained using a force plate, a xylophone plate, electromyography and electrogoniometry. The equipment and methods used are described. The results suggest that the locomotion mechanism in patients with disease of the knee joints is affected adversely by severity of pain and instability. Alterations observed in the gait characteristics of such patients comprise decreased sagittal plane knee motion during stance phase, increased electromyographic activities of the quadriceps and the hamstrings during stance phase and increase in the lateral component of ground reaction force. Functional requirements of the knee joint include stable support of body weight and a wide range of movement. These features make the knee particularly susceptible to the deleterious effects of traumatic injury, inflammatory and degenerative disease. The purposes of this study are to characterise the gait patterns of patients with such disease of the knee joints and to identify the mechanisms of these abnormal patterns. With these intentions, biomechanical analyses were carried out on information obtained from the use of force plates, electrogoniometry and electromyography.


Effect of molecular structure on bile acid-induced alterations in absorptive function, permeability, and morphology in the perfused rabbit colon. An in vivo intestinal perfusion system was used to study the effects of different bile acids on fluid secretion, mucosal permeability, and mucosal morphology in the rabbit colon. To define the structure-activity relationships of the bile acids, nine unconjugated bile acids were used, varying only in the number (two or three) or position (3, 7, or 12 or various combinations) of hydroxy or keto nuclear substituents. Results showed that bile acids with two hydroxy groups in the alpha configuration at the 3,7 position, 3,12 position, or 7,12 position induced fluid secretion, increased mucosal permeability, and produced mucosal damage as assessed by light and scanning electron microscopy and quantitated by DNA loss during perfusion. Replacement of hydroxy groups by keto groups or a change from alpha to beta configuration for the hydroxylic substituent in the 7 position abolished all three activities. Trisubstituted derivatives, whether hydroxy or keto, did not affect fluid secretion permeability or cause mucosal damage. These studies indicate that of the major primary and secondary bile acids in man, only deoxycholic and chenodeoxycholic acids alter colonic structure and function in the rabbit. They show further that the cathartic effects of bile acids have specific structural requirements; and they show that bile acid-induced secretion was invariably associated with increased mucosal permeability and epitheliolysis.


Factor VIII-related antigen in human blood platelets: localization and release by thrombin and collagen. Human platelets contain 0.61 +/- 0.06 (S.E.) U of VIIIR:Ag per 10(9) platelets or about 25% of the circulating antigen in whole blood. Assay of subcellular fractions obtained by sucrose density-gradient centrifugation of disrupted platelets indicates that over 85% of the VIIIR:Ag is in the alpha granule fraction with other nonenzymatic proteins which can be released by treating platelets with collagen or thrombin. Seven percent or less is in the membrane fraction. After exposure of platelet suspensions to an optimal concentration of collagen, the supernatants contain, at most, 30% of the total amount of VIIIR:Ag in the platelets but a higher percentage of other releasable substances. After exposure to thrombin, the supernatants contain virtually no VIIIR:Ag. These low values are attributed to release of a limited amount of VIIIR:Ag by collagen or thrombin rather than to adsorption or destruction of antigen, for several reasons. (1) VIIIR:Ag can be recovered in the platelet residue as effectively as in control samples. (2) The antigen cannot be detected on the surface of collagen- or thrombin-stimulated platelets. (3) Relatively little released antigen is adsorbed by collagen or destroyed by thrombin. Release of VIIR:Ag by collagen is markedly reduced by aspirin, and release of the antigen as well as fibrinogen from platelets is much slower than release of 14C-serotonin and beta-TG.


Zinc metabolism in uremia.

Zinc metabolism in uremia. Subnormal plasma zinc levels have been reported in uremic patients. However, detailed studies regarding zinc status in uremia are not available. Twenty-five patients with chronic renal failure (10 undergoing maintenance hemodialysis, five receiving chronic peritoneal dialysis, and 10 nondialyzed azotemic patients) had lower concentration of zinc in plasma, leukocytes, and hair as well as increased plasma ammonia and ribonuclease activity compared to age- and sex-matched controls (p less than 0.001). Similar biochemical changes have been reported in experimentally induced zinc deficiency in both animals and man, except that erythrocyte zinc concentration was elevated in these patients. High erythrocyte zinc concentration may be related to ineffective erythropoiesis in uremia. The results of this study suggest that abnormality in zinc metabolism occurs commonly in patients with chronic renal failure and that it develops prior to initiation of dialysis treatment.


Endogenous erythroid colony-forming cells in fetal and newborn sheep. In plasma clot cultures fetal, newborn, and adult sheep bone marrow cells respond to Ep, giving rise to large numbers of colonies of erythroid cells. In fetal and newborn, but not adult, sheep the marrow produced EEC in the absence of added Ep. The EEC-forming cells represent a transitory population of erythroid progenitors, persisting for approximately 2 month post-partum. The activity of these cells in the normal newborn sheep displayed a cyclic pattern, which was abolished in the presence of high circulating Ep levels. In both fetal and newborn sheep, EEC formation was inhibited by prior neutralization of Ep present in culture media with anti-Ep, suggesting that as in patients with PV the proliferation and/or differentiation of EEC-forming cells of sheep is subject to Ep control.


Augmentation of peritoneal mass transport by dopamine: comparison with norepinephrine and evaluation of pharmacologic mechanisms. The effect of catecholamines on transport during peritoneal dialysis was studied in unanesthetized rabbits. Intravenous I-norepinephrine consistently decreased peritoneal clearances of urea and creatinine to 84% of control values or less but did not affect osmotically induced water flux. Comparable pressor doses of dopamine increased clearances of urea and creatinine to 145% of control values, whereas osmotic fluid flux increased only slightly. Dopamine also increased urea transport when administered intraperitoneally. The augmentation of solute transport by dopamine was unaffected by simultaneous administration of propranalol, was decreased by phentolamine, and was abolished by haloperidol. Dopamine may be preferable toI-norepinephrine when vasopressor therapy is required during peritoneal dialysis. The augmented transport with dopamine appears to depend on the action of dopamine receptors causing mesenteric vasodilation and in part on alpha-adrenergic receptors simultaneously increasing blood pressure while mesenteric blood flow is maintained.


Improved gas chromatographic quantitation of breath hydrogen by normalization to respiratory carbon dioxide. A gas-solid chromatographic system using tandem silica gel and molecular sieve columns is described for the measurement of hydrogen, carbon dioxide, oxygen, and nitrogen in samples of respiratory gases. This system has a detection limit of 2 ppm of hydrogen in a 1 ml sample and can measure 120 ppm H2 and 5% CO2 with relative standard deviations of 1.3% and 1.7%, respectively. Improved sample storage and withdrawal techniques are described that give reproducible values for up to 6 weeks after collection. Finally we show that normalization of breath hydrogen values to an alveolar concentration, using the observed carbon dioxide concentrations, substantially reduces the range and variance of apparent H2 concentrations in human subjects. Normalization eliminates the need for rebreathing or end-expiratory collection techniques and substantially increases the reliability and clinical utility of hydrogen breath measurements in noninvasive tests of carbohydrate malabsorption.


Effects of sodium loading and depletion in normotensive first-degree relatives of essential hypertensives. To examine potential mechanisms responsible for the greater prevalence of hypertension among relatives of hypertensives than among relatives of normotensives, we subjected 43 normotensive first-degree relatives of known essential hypertensives and 43 age-, race-, and sex-matched normal subjects with no family history of hypertension to sodium loading and depletion. The data show that the relatives had higher blood pressures than did controls. They also had higher PRA values before and after a 2 L intravenous saline infusion over 4 hr. In addition, the relatives excreted less (p less than 0.05) sodium on the day of the infusion than did the controls. PA values and UNe excretion in the two groups did not differ. In the relatives, age correlated with systolic and diastolic blood pressure. Age correlated inversely with the 24 h urine sodium excretion on the sodium loading day. These correlations were not observed in controls. Normotensive first-degree relatives of hypertensives differ from relatives of nonhypertensives in that they have higher PRA values and a blunted natriuretic response to a saline load. These features may predispose them to the development of hypertension.


The forms and transport of plasma cobalamins in normal man and in myeloproliferative states. Seven patients with MP and four controls were injected with 0.04 to 0.05 microgram of [57Co] cyanocobalamin i.m. or i.v. in order to study the shift of binding with time and the conversion of one Cbl to another. The initial pattern of binding reflected the proportion of the apo forms of TC II and R binders. In MP there was more initial binding of the injected Cbl to R-type binders and less to TC II. During the first 48 hr after intake, the injected Cbl remained mostly as cyanocobalamin. Some adenosylcobalamin appeared transiently in both control and MP subjects. Radioactive methylcobalamin did not appear in the circulation until after 48 hr, and the conversion of cyanocobalamin to methylcobalamin within the circulation was greater in MP subjects. Serum from some subjects converted small amounts of cyanocobalamin to all other forms in vitro by a heat-stable, extracellular property which was abolished by dialysis of the serum. This property of serum could have accounted for the early conversion to adenosylcobalamin but not, at least as an in vitro phenomenon, to the late appearance of methylcobalamin. Although the expected increases and abnormal patterns of R binders were observed in MP, these abnormalities could not be correlated with the increased conversion to plasma methylcobalamin. During the first 6 hr after injection the R binder-Cbl designated as alpha 2-R-Cbl disappeared from the circulation at a rate faster than that of alpha 1-R-Cbl. Subsequently the alpha 2 and alpha 1 components of R binder-Clb cleared at the same rate, and this rate was the same for both control and MP subjects.


Characteristics of the dextran-coated charcoal assay for estradiol receptor in breast cancer preparations. The measurement of E2 receptor (E2R) in human breast cancer cytosol is significantly influenced by conditions usually employed in the dextran-coated charcoal assay. The incubation time and temperature have an influence on the rate of binding and stability of the receptor. Since lower temperatures preserve the integrity of the receptor, a 2 hr incubation at 4 degrees was selected as the standard incubation procedure. These conditions allow for the detection of at least 80% of the E2R. With supernatants from high-speed centrifugation of HBT biopsies or the human breast cancer cell line MCF-7, reducing agents increased the apparent E2R binding in the order: DTT greater than G-SH greater than MTG. The maximum enhancement of specific E2R binding by a given thiol agent was dependent on its concentration in the incubation medium. The optimum DTT level (7.5 mM) for MCF-7 cell homogenization and cytosol equilibration with tritiated E2 increased E2R to two times control (no DTT). For the HBT 150,000 g supernatant, 1 mM DTT was required to optimize the E2R quantitation. The duration of the dextran-coated charcoal extraction of the cytosol-[3H]E2 incubation had no effect on the level of E2R up to 21 hr. Minimum levels of nonspecific binding of [3H]E2 could be obtained after 4 hr extraction. Maximum depletion of specific [3H]E2 binding could be obtained by adding between 200- and 1000-fold molar excess of unlabeled E2. Greater amounts of unlabeled steroid displaced the radioactive E2 from the dextran-coated charcoal, thereby artifactually increasing the apparent nonspecific binding. This phenomenon may be overcome by utilizing more dextran-coated charcoal in the extraction. However, there was a 9% loss of specifically bound [3H]E2 per milligram of dextran-coated charcoal (1:10 dextran to charcoal by weight) when the cytosol protein was below 90 microgram per incubation. Supplementation with 200 microgram or more albumin per incubation prevented this loss. The dextran:charcoal ratio also prevented E2R loss in the order: 1:1 greater than 1:10 greater than 1:100. One milligram of dextran-coated charcoal (1:10) has the capacity to adsorb 0.3 to 0.4 microgram of free E2. Other unlabeled competitors are capable of displacing [3H]E2 on the receptor. Although DES was as effective as E2, U11,100A and estrone were inefficient competitors. It appeared that the levels of these two estrogen analogues required to maximally displace [3H]E2 on receptor also eluted labeled E2 from the dextran-coated charcoal. DES, however, was unable to displace significant quantities of the [3H] E2 from dextran-coated charcoal even at a molar excess of 50,000:1.


Tyrosine metabolism in cirrhosis.

Tyrosine metabolism in cirrhosis. The aim of this study was to define the enzyme defect responsible for tyrosinemia in cirrhotic patients. The principal hepatic degradation pathway for tyrosine, tyrosine leads to p-hydroxyphenylpyruvic acid equilibrium homogentisic acid leads to CO2 was studied in 18 cirrhotic patients and eight controls. The classic method employed in elucidation of hereditary tyrosinosis was sued. Metabolic intermediates on the pathway were measured in the basal state, and following oral loading doses (50 mg/kg BW) of tyrosine, PHPA, and homogentisic acid. Cirrhotic patients showed a significant increase (p = 0.005) in fasting plasma tyrosine and in basal PHPA excretion and impaired tolerance to all three metabolites when compared to normals. Fifteen of the 18 cirrhotic patients showed tyrosine intolerance which was not accompanied by change in distal metabolites compared to their basal levels. Nevertheless 13 of the 18 did exhibit intolerance of either PHPA or homogentisic acid. We conclude that in contrast to the single complete defect in hereditary disorders of tyrosine metabolism, cirrhotic patients have partial defects at tyrosine transaminase, PHPA oxidase, and homogentisic acid oxidase, the initial step being rate-limiting.


Catabolism of low-molecular-weight hydroxyethylated amylopectin in man. I. Changes in the circulating molecular composition. Intravascular persistence concomitant with changes in the circulating molecular composition were determined in six fasted normal men dosed with 400 ml of 14% LMW-HES (a new plasma expander). The concentration of LMW-HES in serum fell to half its peak value in 3.9 +/- 1.1 (S.D.) hr, whereas serum levels of glucose remained elevated throughout the 12 hr postinjection fasting period. The LMW-HES recovered from the intravascular space was shown by gel filtration on a column of Sepharose CL-4B to be a narrower molecular size distribution (less polydispersion) than the injected material. The ratio of Kav . urine/Kav . injected solution was 1.34. At 30 min after injection, however, the ratio of Kav . urine/Kav . serum was 1.20, and by 24 hr, the value was 1.15. Overall, changes in the molecular distribution in the bloodstream between the end of the infusion period and 24 hr later were small. The results suggest that the intravascular catabolism of LMW-HES may occur in two distinct phases: a rapid initial degradation, followed by a more gradual elimination influenced by the MS of the injected material.


Studies on the absorption of desmosine and isodesmosine. Intestinal absorption of desmosine and isodesmosine was investigated in several animal species. Hamsters given desmosine injections intraperitoneally excreted 70% of the injected dose in the urine. When desmosines or elastin were given via gastric intubation, desmosines failed to appear in the urine, indicating a lack of desmosine absorption. Additional studies with everted hamster intestine in vitro showed that norleucine was actively transported from the mucosal to the serosal side but that desmosines and lysinonorleucine were not transported. Intestinal slice experiments with hamster, rabbit, and dog intestine indicated desmosines only in the extracellular space and not absorbed by the mucosal cells. These studies strongly suggest that desmosines are not absorbed via the gastrointestinal tract. The importance of this study rests with the conclusion that desmosines recovered in the urine must represent the degradation products of elastin destruction in the tissue. Newly synthesized elastin cannot interfre, since elastin precursor molecules do not contain desmosines. Thus quantitation of urinary desmosines may be of clinical value as a method of evaluating elastin catabolism.


Bronchoalveolar cell populations in acute sarcoidosis. Observations in smoking and nonsmoking patients. Bronchoalveolar cells were obtained by segmental lung lavage from 14 patients with sarcoidosis and studied to determine changes in composition associated with acute disease and smoking history. In nonsmoking patients, both the proportion and number of lymphocytes recovered were significantly increased compared to nonsmoking controls (p less than 0.001 in both cases). Furthermore, the number of T cells was increased (4.19 +/- 0.72 vs. 0.896 +/- 0.107 x 10(6) cells/100 ml, p less than 0.001), and activated T cells were also increased (1.04 +/- 0.478 vs. 0.148 +/- 0.032 x 10(6) cells/100 ml, p less than 0.01). Less striking differences in these values were found when smoking sarcoid patients were compared to smoking controls, but there were no significant differences in these values for smoking and nonsmoking patients (p greater than 0.05). Thus acute sarcoidosis in both smoking and nonsmoking patients appears to be associated with a lymphocytosis of the bronchoalveolar space which results from increased numbers of T cells and activated T cells. The potential importance of this finding with regard to diagnosis and pathogenesis is discussed.


The effect of chronic uremia on portal and systemic ammonemia in normal and portal-strictured rats. It is commonly assumed that the uremia occurring in cirrhotic patients leads to hepatic encephalopathy by means of hyperammonemia produced by bacterial hydrolysis of urea in the digestive tract. Therefore the effect of an increase in circulating urea on portal and systemic blood ammonia levels was investigated in normal rats and in rats with a strictured portal vein. It was shown that moderate chronic uremia, produced by subtotal nephrectomy, significantly increases both portal blood ammonia levels in normal rats and the systemic hyperammonemia resulting from portosystemic shunts in portal-strictured rats. In the latter group, despite a prolonged systemic hyperammonemia of around 400 micrograms/dl, no obvious behavioral modification could be observed, but a significant slowing of the growth rate occurred. This slowing, however, was not correlated with the systemic blood ammonia levels.


Decreased hepatic microsomal reserve in patients with cirrhosis. Studies using aminopyrine as model drug. It is unclear whether hepatic drug metabolism which is decreased in patients with liver disease, can be stimulated by enzyme-inducing drugs. Hepatic microsomal reserve, defined as the difference between the basal and phenobarbital-stimulated ABT, was therefore studied in eight healthy control subjects and 12 patients with stable alcoholic cirrhosis. The ABT increased significantly (p less than 0.01) from a basal value of 6.1% +/- 0.8 (mean +/- S.D.) to 8.9% +/- 0.8 in the eight control subjects after phenobarbital ingestion. In the 12 patients with alcoholic cirrhosis the basal ABT was 2.9% +/- 1.5 and did not change significantly after phenobarbital ingestion, when the value was 3.0% +/- 1.6. A small increase in the ABT occurred after phenobarbital ingestion in five of the patients with alcoholic cirrhosis, but in no patient did this increase bring the ABT within normal limits. We conclude that in many patients with stable alcoholic cirrhosis aminopyrine metabolism is decreased and cannot be corrected by treatment with phenobarbital.


Epidemiologic interference of virus populations. There are a few simulation studies for interference models in the literature but the present paper discusses an analytical model for the competition of two interfering virus populations in a community. The mathematical model consist of eight coupled differential equations which have up to four equilibrium points. Criteria for local stability are given.


Serial learning in the pigeon.

Serial learning in the pigeon. Three pigeons learned to peck four colors in a particular sequence, regardless of how these colors were positioned on four response keys and without feedback following each response. This demonstrates that serial learning is possible in subprimate animals.


Separating the reinforcing and discriminative properties of brief-stimulus presentations in second-order schedules. Pigeons' responses were maintained under multiple schedules to study properties of briefly presented stimuli. Responses in one component produced food according to a second-order schedule with fixed-interval components in which food or a brief stimulus occurred with equal probability. In the second component responses produced only the brief stimulus under a fixed-ratio schedule. Under various conditions the brief stimulus in the first component was (a) paired with food, (b) not paired with food, (c) partially omitted, or (d) scheduled simultaneously with the second-order schedule under an independent variable-interval schedule. Paired and nonpaired brief stimuli maintained similar response patterning in the second-order schedule. However, only paired stimuli maintained responses in the second component. The data suggest that nonpaired brief stimuli engender response patterning in second-order schedules as a result of their discriminative properties. When the stimulus is paired with food, these discriminative properties sometime mask a reinforcement effect, and no change in response patterning is observed. When the discriminative properties of the brief stimulus are absent, the reinforcing effects of pairing the brief stimulus with food may be observed.


Auditory discrimination: role of time and intensity in the precedence effect. Rats were trained to respond on a lever adjacent to a sounding speaker (the sound source) when a single click was emitted. A second click (the artificial echo) was presented through a second speaker on the opposite side. In Condition I, the echo (equal in intensity to the source) was delayed from .015 to 32 milliseconds; greater than 75% correct responses were given for delay times between about .040 milliseconds (lower threshold) and 8 milliseconds (upper threshold). In Condition II, the echo (simultaneous with the source) was reduced in intensity relative to the source over a range from 2.5 decibels to 40 decibels; greater than 75% correct responses occurred for intensity reductions greater than 5 decibels. In Condition III, both the intensity and the delay time of the echo were manipulated in a manner analogous to that which would occur under natural conditions; greater than 95% correct responses were given for delay times from 1 to 32 milliseconds. These data indicate that both time and intensity differences are necessary for localization of primary sources, with delay time contributing more at short echo path distances, and intensity differences at long distances.


The control of responding by sounds: unusual effect of reinforcement. Naive rats were trained to respond on one lever in the presence of noise bursts from one speaker and on a second lever in the presence of noise bursts from a second speaker. The speakers were mounted behind the levers. When responding on the lever adjacent to the sounding speaker was reinforced, control developed within fewer than five trials. When responding on the nonadjacent lever was selectively reinforced, responding on the lever adjacent to the sounding speaker increased in probability for several sessions. Naive rats were trained to respond on the nonadjacent lever following preexposure to the sound. Responding on the lever adjacent to the sounding speaker increased in probability, showing that novelty was not responsible for the effect. Naive rats were run on automaintenance procedures in which there was no explicit pairing of sound and magazine operation, 100% pairing of sound and magazine operation, or magazine operation following 40% of sound presentations. None of the rats acquired the response of approaching and sniffing the sounding speaker, indicating that sound-magazine pairing was not responsible for the effect.


Matching-to-sample accuracy on fixed-ratio schedules. Pigeons performing on a matching-to-sample procedure were exposed to six fixed-ratio (FR) schedules (FR 1, 5, 10, 20, 40, and 60) of food reinforcement for correct matching responses. During both a correction and a noncorrection procedure without an intertrial interval (ITI), matching accuracy was lower on FR 1 and FR 60 than at intermediate ratios. With the FR 1 schedule, both a 5-sec and a 25-sec ITI resulted in higher matching accuracy than without an ITI; accuracy, with an ITI, was fairly constant for ratios of 1 to 20 but declined at higher ratios. The results suggest that the presence or absence of an ITI in matching to sample may account for inconsistencies obtained in earlier studies of the relationship of matching accuracy to ratio size.


The allocation of time to temporally defined behaviors: responding during stimulus generalization. In one stimulus condition, reinforcement depended on rats holding a lever for a duration having both minimum and maximum boundaries. During a second light intensity, reinforcement was not available for some rats; for others, reinforcement depended on a second response duration requirement. Generalization test stimuli controlled the same response durations found during training, and the amount of time allocated to a given response duration depended on the proximity of the test stimulus to the training stimulus which controlled that particular duration. The results indicated that a gradient of stimulus control does not reflect an underlying continuous change in responding, but is a result of the mixing of responses previously controlled by stimuli present during conditioning.


Concurrent schedules: undermatching and control by previous experimental conditions. Five pigeons were trained on concurrent variable-interval schedules. A series of conditions in which the ratio of reinforcement rates on two keys was progressively increased and then decreased was arranged twice. The birds were then exposed to an irregular sequence of conditions. Each condition in which reinforcement was available on both keys lasted six sessions. Performance in the first, third, and sixth sessions after a condition change was analyzed. Following a condition change, preference was biased toward the preference in the last condition, but this effect largely disappeared before the sixth session of training. The birds' preferences also appeared less sensitive to reinforcement rates in early sessions after a transition. Preference in a session was a function of both the reinforcements in that session and the reinforcements obtained in as many as four or five previous sessions. The effects of reinforcements in previous sessions could be summarized by the performance in the immediately preceding session, giving a relatively simple relation between present performance and a combination of present reinforcement and prior session performance. While such hysteresis could cause undermatching when only a small number of sessions are arranged in a condition, undermatching in a stable-state performance probably arises elsewhere.


The relationship of the central motor pattern to the feeding cycle of Lymnaea stagnalis. Electromyographic recordings from the buccal muscles of Lymnaea during feeding has shown that there are 4 component phases in the feeding cycle. Cinephotography of feeding cycles has confirmed that these correspond to protraction, 2 phases of retraction, and an inactive phase. The 4 phases of muscle activity can also be related to the cycle of neural activity described previously (Benjamin & Rose, 1979). Thus types 6, 4 group, and type 8 cells are motoneurones involved in protraction and the two retraction phases, while the type 5 cell fires in the inactive period. The combination of physiological and anatomical approaches has led to the suggestion that the single and double input cells described by Benjamin & Rose (1979) are involved with the control of buccal and oesophageal activity respectively.


Matching, undermatching, and overmatching in studies of choice. Almost all of 103 sets of data from 23 different studies of choice conformed closely to the equation: log (B(1)/B(2)) = a log (r(1)/r(2)) + log b, where B(1) and B(2) are either numbers of responses or times spent at Alternatives 1 and 2, r(1) and r(2) are the rates of reinforcement obtained from Alternatives 1 and 2, and a and b are empirical constants. Although the matching relation requires the slope a to equal 1.0, the best-fitting values of a frequently deviated from this. For B(1) and B(2) measured as numbers of responses, a tended to fall short of 1.0 (undermatching). For B(1) and B(2) measured as times, a fell to both sides of 1.0, with the largest mode at about 1.0. Those experiments that produced values of a for both responses and time revealed only a rough correspondence between the two values; a was often noticeably larger for time. Statistical techniques for assessing significance of a deviation of a from 1.0 suggested that values of a between .90 and 1.11 can be considered good approximations to matching. Of the two experimenters who contributed the most data, one generally found undermatching, while the other generally found matching. The difference in results probably arises from differences in procedure. The procedural variations that lead to undermatching appear to be those that produce (a) asymmetrical pausing that favors the poorer alternative; (b) systematic temporal variation in preference that favors the poorer alternative; and (c) patterns of responding that involve changing over between alternatives or brief bouts at the alternatives.


Thermoacclimatory changes in the ionic microenvironment of haemoglobin in the stenothermal rainbow trout (Salmo gairdneri) and eurythermal carp (Cyprinus carpio). 1. Haematological characteristics (erythrocyte number, haematocrit, haemoglobin, mean erythrocyte volume and haemoglobin content) and plasma and packed red blood cell water and electrolyte (Na+, K+, Ca2+, Mg2+, Cl-) levels were determined in summer and winter populations of rainbow trout acclimated to 2, 10 and 18 degrees C, and for carp held at 2, 16 and 30 degrees C. Erythrocyte electrolyte concentrations and ion:haemoglobin ratios were calculated from these data. 2. Modest increases in red cell abundance and reductions in mean erythrocytic volume were the most obvious haematological changes accompanying acclimation to higher temperatures. Haemoglobin levels in carp also tended to increase with temperature. 3. In winter trout plasma sodium and potassium were elevated following acclimation to increased temperature. No significant cges in plasma composition were observed in summer fish. Carp held at higher temperatures were characterized by increases in plasma chloride and calcium and reductions in sodium and magnesium levels. 4. Red cell potassium and magnesium and K+:Hb and Mg2+:Hb ratios tended to be higher in winter than in summer trout, with the converse being true of chloride and calcium and Cl-:Hb and Ca2+:Hb. Only potassium and K+:Hb were significantly altered following acclimation; rising at higher temperatures. In carp, potassium and K+:Hb were relatively thermostable, but sodium and chloride and Na+:Hb and Cl-:Hb increased with temperature while magnesium and Mg2+:Hb decreased. Changes in the ionic composition of carp red cells support the suggestion that cellular pH is reduced in the warm-acclimated animal. 5. These variations may be of adaptive value. Increases in chloride and hydrogen ion commonly reduce haemoglobin=oxygen affinity, and should facilitate oxygen unloading at the tissue level. Reductions in cellular magnesium, by maximizing organophosphate modulator levels, should produce much the same effect. 6. In both species reductions in mean erythrocytic volume took place at higher temperatures despite increases in cellular ion content which exceeded those of plasma. It is probable that reductions in cellular volume, which should favour branchial oxygen loading, were achieved by export of some as yet unidentified solute or solutes.


On the endogenous bursting properties of 'light yellow' neurosecretory cells in the freshwater snail Lymnaea stagnalis (L.). Intracellular microelectrodes were used to study a cluster of neurosecretory 'Light Yellow' Cells (LYC) in the central nervous system of Lymnaea stagnalis. LYC usually have a spontaneous firing pattern of bursts, lasting 10--600 s, alternating with periods of silence. Experiments on isolated single cells showed that the bursting activity has an endogenous origin. Each action potential is followed by a depolarizing afterpotential (DAP), with an amplitude of about 10 mV, lasting several seconds. Bursts end with a subthreshold DAP. It is concluded that two pace-maker mechanisms are responsible for the bursting pr-perties, one initiating and the other (the DAP) maintaining the burst. The relationship between the electrical and the neurosecretory properties of the cells is discussed.


Major histocompatibility complex-restricted antibody reactivity mainly, but not exclusively, directed against cells from male donors. An IgM antibody, present in the serum of a female patient with aplastic anemia, is described that reacted in a modified complement-dependent cytotoxicity test with a subset of the B cells from HLA-A2-positive, but not HLA-A2-negative males. With the exception of two HLA-A2 positive females, the antibody did not react with other cells from either HLA-A2-positive or HLA-2-negative females. The cells of one of these and from HLA-A2-positive males were able to absorb the antibody from the serum. Cells from other donors were unable to absorb the antibodies. The mononuclear cells of the same patient were cytolytic in cell-mediated lympholysis (CML) for phytohemagglutinin blasts from all HLA-A2-positive males and one of the females reacting with antibody, but not with blasts from HLA-A2-negative males and all other females. Thus, the results obtained with the antibody in the complement-dependent cytotoxicity test showed an almost perfect correlation with cytolysis in CML tests. These results suggest that the IgM antibody may be the first example with major histocompatibility complex restriction. Because the antibody reacted with the cells from two female donors, the restricting determinant is not, in all probability, the H-Y determinant.


Nonenzymatic glycosylation, sulfhydryl oxidation, and aggregation of lens proteins in experimental sugar cataracts. The formation of sugar-cataracts has been hypothesized to involve the nonenzymatic glycosylation, sulfhydryl oxidation, and aggregation of lens proteins. Cataractous lenses of diabetic and galactosemic rats were analyzed for glycosylated lysine residues in crystallins. A five- and a ten-fold increase in glycosylated lysine residues was measured in galactose and diabetic cataracts, respectively. The modification was predominant in the insoluble fraction of the lens homogenate. The proteins were further examined for the presence of disulfide bonds and high molecular weight aggregates. After careful disruption of the lens in a nitrogen environment, a cloudy solution was obtained from cataractous lenses whereas a clear solution was obtained from normal lenses. The absorbance at 550 nm of the solution of both the galactosemic and the diabetic cataracts could be decreased by approximately 50% with the addition of dithioerythritol (50 mM). The presence of high molecular weight aggregates was ascertained by sucrose gradient centrifugation and gel filtration chromatography. The proteins were heterogenous in size and showed a mol wt range of 36 to greater than 176 million daltons. Treatment with dithioerythritol induced a marked decrease in the amount of high molecular weight proteins. These data suggest that sugar cataracts of experimental animals have, in common with human cataracts, the presence of high molecular weight aggregates which are in part linked by disulfide bonds.


The autoactivation of rabbit Hageman factor. Proteolytic cleavage and activation of isolated, single chain, zymogen Hageman factor was observed in the presence of kaolin alone. The rate of cleavage of kaolin-bound Hageman factor was enhanced 50-fold by the presence of prekallikrein and high molecular weight kininogen. The two-chain 82,000 dalton form of activated Hageman factor (alpha-HF(a)) also cleaved kaolin- bound single-chain Hageman factor in a dose-dependent manner, yielding fragments of 28,000 and, 50,000 dahons under reducing conditions. Cleavage of kaolin-bound single-chain Hageman factor was not inhibited by preincubation with diisopropylfluorophosphate (12 mM) for 10 min, but long-term incubation of Hageman factor with diisopropylfluorophosphate (up to 48 h) resulted in inhibition of cleavage of kaolin-bound Hageman factor to an extent proportional to the inhibition of procoagulant Hageman factor activity. Hageman factor cleavage was maximal when the kaolin concentration was {approximately} 10-fold greater than the Hageman factor concentration (wt:wt), and was partially inhibited by high molecular weight kininogen. Kaolin-bound Hageman factor cleaved clotting factor XI in an amount which correlated with the extent of cleavage of the Hageman factor. These findings are compatible with the concept that single-chain Hageman factor and alpha- HF(a), are both capable of cleaving and activating kaolin-bound Hageman factor and that a close molecular association of kaolin-bound Hageman factor molecules is required for this reaction.


Tumorigenicity of Indian muntjac diploid cells by the proviral integration of sarcoma gene of a mouse retrovirus. The transformed clonal isolates of Indian muntjac diploid cells by a mouse sarcoma virus, 43-2XV, were tested for tumorigenicity in athymic nude mice. In spite of the indistinguishable transformed morphology, the tumorigenicity exhibited four different patterns: (a) no tumor formation; (b) slowly growing regressive tumor formation; (c) rapidly growing regressive tumor formation; and (d) rapidly growing progressive tumor formation. This demonstrates that the same diploid host cells transformed by the same virus reveal variable patterns of tumorigenic expression and some transformed host cells lack the tumorigenicity entirely. The findings that there are at least two chromosomes and four recombinant sites assigned for the proviral integrations of the sarcoma gene into the Indian muntjac gene (M. Hatanaka, R. Klein, R. Kominami, T. Oikawa, H. Okabe, N. Tsuchida, E. C. Connors, and A. Carrano. Transformation of Indian muntjac diploid cells by the proviral integration of sarcoma gene of a mouse retrovirus. Manuscript in preparation.) lead us to propose a hypothesis that variable expressions of tumorigenicity under the neutral background of immune responses, may arise from variable integrations of the sarcoma gene into the host chromosome.


[The calculation of hemoglobin concentration from blood and plasma densities, measured by the mechanical oscillator technique (author's transl)]. Blood samples from patients were taken for the measurement of blood and plasma densities by the mechanical oscillator technique, and the hematocrit value and the hemoglobin content of the blood. The values for blood and plasma density permit the precise calculation of blood hemoglobin concentration and hematocrit. Thus, in addition to the calculation of total protein concentration of plasma (Holzer et al (1978) J. Clin. Chem, Clin. Biochem. 16, 391--395), measurement of the blood density yields two further relevant parameters. In total, only 3 ml of blood are needed for the measurements.


Surface-associated heparin inhibits zymosan-induced activation of the human alternative complement pathway by augmenting the regulatory action of the control proteins on particle-bound C3b. Discrimination by the human alternative pathway between activating and nonactivating particles occurs after deposition of C3b by the continuous low-grade interaction of the alternative pathway components in biologic fluids and is dependent on the modulation by surface constituents of the interaction of bound C3b with the control proteins, beta 1H, and C3b inactivator (C3bINA). When heparin glycosaminoglycan was coupled to activating particles, such as zymosan or Sepharose, by cyanogen bromide activation, their capacity to activate the human alternative pathway was inhibited. The loss of alternative pathway-activating capacity was directly correlated to the number of heparin molecules bound/zymosan particle, whether the ratio was varied by increasing the amounts of heparin in the initial coupling reactions or by treating a fully inhibited particle with incremental concentrations of heparinase. Analysis by linear regression of the inhibitory effect of each procedure (r = 0.97, r = 0.98, respectively) for adjusting the number of heparin molecules/particle revealed that the dose-response relationships were identical and that complete inhibition occurred with greater than 12 X 10(8) molecules of heparin/zymosan particle. The coupling of heparin to zymosan did not impair the uptake of C3b from the fluid-phase interaction of C3, B, and D, and did not alter the capacity of bound C3b to associate with B so as to permit its inactivation by D. Although the regulatory proteins present in normal serum chelated with EDTA or presented as a combination of purified C3bINA and beta 1H were relatively inefficient in inactivating C3b function on an activating particle of the alternative pathway such as zymosan or zymosan-cyanogen bromide, the control proteins rapidly inactivated C3b on a nonactivating particle wuch as a sheep erythrocyte or zymosan with coupled heparin. The increased numbers of C3b sites susceptible to inactivation by C3bINA in the presence of beta 1H were significantly correlated to the number of molecules of heparin/particle. By linear regression analysis of the correlation (r = 0.99) the number of heparin molecules/particle required to promote total inactivation of bound C3b by purified control proteins was 13.8 X 10(6). This molecular analysis suggests that the action of heparin coupled to an activating particle of the alternative pathway is to promote the interaction between particle-bound C3b and the regulatory proteins, thereby preventing particle-associated amplified C3 cleavage. It is noteworthy that both surface constituents known to maintain a particle as a nonactivator of the alternative pathway, sialic acid and N-sulfated mucopolysaccharide, act by facilitating the inactivation by regulatory proteins of the function of particle-bound C3b.


Bromocresol purple, a non-specific colour reagent for the determination of serum albumin. The determination of serum albumin with the dye reagent bromocresol purple was investigated. We found that bromocresol purple is not a specific reagent for albumin, but that serum proteins in the alpha-, beta-, and gamma-globulin fractions also react with this dye. Similar results were obtained for bromocresol green. Furthermore there is a great difference in absorbance between human and bovine albumin solutions (having the same concentration) when using the bromocresol purple reagent. Most probably this is a result of the difference in interaction of the reagent with the substrate. Although the standard curves with the bromocresol purple method were more linear (especially at higher albumin concentrations) than those obtained with the bromocresol green method, we still prefer the bromocresol green technique to the bromocresol purple technique, because its results are more comparable with those of other methods. Moreover in quality control and calibration procedures, strong deviations from the recommended values are obtained when non-human sera are analyzed with the bromocresol purple method. For reasons of accuracy more specific methods are preferable.


Evaluation of a new diagnostic kit for the enzymatic determination of creatinine. We have evaluated a new diagnostic kit for the enzymatic determination of creatinine in serum, plasma and urine. The method was found to be linear up to 1000 mumol/l. Within-run precision had a coefficient of variation of 3.1--5.6% for normal to slightly elevated values and of 2.3--3.3% for values of about 320 mumol/l. Day-to-day precision was found to be 7.8 and 5.7% for values of 94 and 141 mumol/l, respectively. Mean analytic recovery was 98%. Bilirubin and hemoglobin did not interfere in concentrations up to 490 and 240 mumol/l, respectively. The influence of lipids was studied. Correlation studies were done with two other methods, the Technicon SMA 12/60 method and the Dutch standard manual method.


[Comparison of ion-selective electrodes and flame photometry for the determination of serum Na+ and K+ for clinical purposes (author's transl)]. The ORION SPACE-STAT (SS-30) and the TECHNICON STAT/ION were used to investigate quality control of Na+- and K+-determinations in test sera (n=8) and in the plasma of 100 patients. The flame photometer IL 543 was used as a reference apparatus. The ion-selective electrode instruments, SS-30 and STAT/ION, gave results very similar to those of flame photometry. The values from the SS-30 were higher by an average of 3% to 5% compared with the values from the other instruments. An explanation for this bias may be the fact that the SS-30 (unlike the STAT/ION and the IL 543) measures electrolyte activities or concentrations in undiluted plasma water. On the basis of these results the routine use of ion-selective electrodes for Na+- and K+-determination can be recommended for clinical laboratories.


Comparative evaluation of different spectrophometric methods for the determination of small amounts of carboxyhaemoglobin. Four spectrophotometric methods measuring HbCO were investigated in comparison with an analyzer (CO-Oximeter) in order to find a method determining HbCO fractions) below 0.10 with sufficient reliability. The CO-Oximeter was chosen as "reference" method because results of other authors with this instrument yielded good agreement with other methods, e.g. IR-spectrometry, GLC, van Slyke. Only the five wavelength method of Commins & Lawther (1965) Brit. J. Industr. Med. 22, 139--143) can be recommended, for it yielded CV's and sensitivities comparable to the CO-Oximeter. The within-run precision had CVs of 2.3% to 4.7%, the sensitivity was 0.0021 to 0.0028. The method is suited even for HbCO measurements of shorter series.


[Evaluation of the aca lipase (DuPont) determination and of the lipase determination according to Myrick (Calbiochem-Behring) (author's transl)]. Two lipase methods, the method for the aca (DuPont) and the method according to Myrick (1976) Thesis, Birmingham Al.) (Calbiochem-Behring), were compared with the lipase method of Rick (1969) Z. Klin. Chem. Klin. Biochem. 7, 530--539) and with the amylase determination. The diagnostic value of the aca method, the amylase and the lipase method of Rick were in good agreement, whereas the method of Myrick gave conflicting results.


[Falsely positive values in multi-channel analysis: An iniquiry into reference and patient groups (author's transl)]. The number of falsely positive values occurring in 12-channel analysis was determined in two groups of patients and reference individuals. It revealed that the portion of falsely positive values actually found was statistically significant beyond that calculated on the assumption of a binomial distribution. Partly distinct correlations of the parameters combined to a profile as well as clear deviations from the normal distribution have to be taken into consideration as reasons for this discrepancy between theory and reality. The results show that the application of the binomial distribution leads to statements which significantly differ from the conditions actually present.


A semi-automated Bratton & Marshall micromethod for determining acetylator phenotype of rabbits using the Abbott Bichromatic Analyzer-100. A micromethod for the determination of acetylator phenotype using the method of Bratton & Marshall (1939) J. Biol. Chem. 128, 537--550) was adapted to the Abbott Bichromatic Analyzer (ABA-100). Advantages of this procedure include: (a) the requirement for small blood samples (20 microliter) making it a suitable pediatric method and eliminating the need for venipunctures; (b) elimination of tedious manual reading of absorbances of samples and mathematical manipulation of data by the automated capabilities of the ABA-100; (c) an increase in the accuracy of the assay by reading samples at two wavelengths thereby correcting for differences in background.


Platelet counting with a laser nephelometer. A method for platelet counting is described, based on the Laser nephelometric principle. Experimental results are reported, together with the practical considerations for the standardisation and correlation of the method, and for application of the method in the routine biochemical laboratory. Venous blood, taken with EDTA-NaCL solution according to Schulz et al (1071, Z. Klin. Chem. Klin. Biochem. 9, 329-333) is used. The blood is centrifuged at 100 g for 10 min and 10 microliter of the supernatant is added to 3000 microliter of a suspending medium (dilution 1:80); 300 microliter platelet suspension are read in a nephelometer cuvet or tube against blank. The number of platelets per liter blood are determined with the aid of a standard curve. The sensitivity and reproducibility of the method, and the correlation with the "electronic coulter counting" method are satisfactory.


Two-dimensional electrophoretic analysis of ribosomal proteins from chronically injured liver. Proteins of the small and large ribosomal subunit, isolated at various times from long-term thioacetamide-damaged rat livers, were analysed by two-dimensional polyacrylamide gel electrophoresis and compared with those from normal liver. Chronic hepatic injury induced a number of time-dependent changes of the structural proteins of the small subunit, whereas the proteins of the large subparticle were essentially unaffected. The most significant alterations were an anodical dislocation of protein S6, a strong diminution in the amounts of proteins S9 and S10 and the occurrence of 3 to 4 additional small subunit proteins. By autoradiographic studies it was established that the modifications of S6 were brought about by an enhanced phosphorylation of this protein, which was the earliest sign of a ribosomal alteration in injured liver tissue.


Enzymic assay of plasma cholesterol: a comparison of analytical variations found using the Greiner GSA II and the Technicon SMA 12/60 and SMA II. Cholesterol was assayed on the Greiner GSA II and the Technicon SMA 12/60 and SMA II autoanalyzers, with Allain's entirely enzymic method (Allain et al. (1974), Clin. Chem. 20, 470--475). The coefficients of variation in the results over a period of one month were less than 6% with the GSA II and the SMA II, but up to 9% on the SMA 12/60. Although the accuracy of the enzymic method depends on the specificity of the cholesterol oxidase and cholesterol esterase, the results were in good agreement with those obtained by Abell's method (Abell et al. (1952), J. Biol. Chem. 195, 357--366) on fresh or freeze dried human plasma. The cholesterol concentration was stable at 4 degrees C, or in frozen or freeze dried plasma. None of the drugs tested interfered with the enzymic assay.


Comparison of 9 methods for the determination of cholesterol. Seven enzymatic procedures for the determination of cholesterol in serum were compared with the Liebermann-Burchard- and a gas-chromatographic method. Using a decision matrix all methods could be ranked according to reliability and practicability . With the exception of the cholesterol oxidase-coupled Kageyama principle and the Liebermann-Burchard procedure, all the other methods showed similar reliability.


[Falsely positive values in multi-channel analysis: an estimation of their frequency with Sylvester's formula and their reduction by a multivariate test quantity (author's transl)]. The application of the binomial distribution for determining the number of "falsely positive values" is not suitable because of the partly strong correlations between the clinical chemical parameters. We have therefore tried to obtain more precise estimations for the number of falsely positive values by using a generalisation from Sylvester's formula and taking into account the correlations. We use multivariate statistics for testing a patient's value" when several laboratory parameters are simultaneously considered. The multivariate technique leads firstly to a significant reduction of the erroneously interpreted "falsely positive values" and secondly allows the detection of hidden implausible constellations of values.


[Continuous intraoperative measurement of Na+, K+ and Ca++ with carrier membrane disc electrodes (author's transl)]. The application of a two-channel flow-through analyzer with carrier membrane disc electrodes for the electrochemical analysis of heparinized blood, plasma or serum is described, its capabilities discussed and results of direct patient measurements presented.


[Calculation of the IgG fraction of cerebrospinal fluid locally synthesized in the central nervous system (author's transl)]. The cerebrospinal fluid (CSF)/serum concentration ratios of immunoglobulin G and albumin were determined for a group of n = 334 control patients. The correlation coefficient relating the two quotients was r = 0.71. The regression line was found to be y = 0.41 x + 0.00014. For statistical and functional reasons it is plausible that the theoretical line should go through the origin. The ocrresponding function would than be y = 0.43 x. The confidence range of the IgG quotient (y) for a given albumin quotient (x) is characterised by +/- 2 Sy.x = +/- 0.001 (Sy.x is the standard deviation of the y values from the regression line). If the IgG quotient is evaluated for the individual albumin ratio of the patient, the following formula can be used for calculation of the locally synthesized concentration of pathological IgG (IgGp) in CSF: IgGp = IgG(CSF) - (0.43 Alb(Serum)-Alb(CSF) + 0.001) . IgG(Serum). This method of determination is independent of the variables which are known to influence the individe, sex, individual blood brain barrier condition, extraction volume of CSF and the method used for protein determination. In comparison with other methods, particularly with respect to the statistics and biochemistry, the reported formula allows an optimal evaluation of pathological IgG values in CSF.


A simple, automatic means of designing a conversion table for expressing laboratory results in the International System of Units (SI). A simple, automatic means of designing a conversion table for expressing laboratory results in the international system of units (SI) is described. The increasing use of the international system of units for expressing the results of clinical chemistry and haematology sets doctors, biologists and paramedical workers the problem of converting the results from the previous confused system of units into the SI system. We propose a simple method for establishing a conversion scale from one system to the other, irrespective of the parameter. This is based on a Hewlett-Packard 9830A calculator equipped with a 9862A plotter. The BASIC language is used. Accordingly we have prepared conversion tables for the most commonly used parameters into the international system of units, on behalf of the Swiss Academy of Medical Sciences.


[Influence of storing conditions on the amino acid concentration in human serum (author's transl)]. Whole blood, serum and deproteinized serum were stored at room temperature up to 24 h and at - 30 degrees C up to one month. The amino acid content was then determined with an automatic amino acid analyser. When whole blood is left at room temperature the concentration of citrulline, alpha-aminobutyric acid, cysteine and tryptophan remains unchanged, whereas the other amino acids show a remarkable increase or decrease. In serum stored for 24 h at room temperature, only the concentrations of aspartic and glutamic acid, serine, cysteine and phenylalanine were altered. With the exception of aspartic and glutamic acid it was possible to leave deproteinized serum up to 24 h at room temperature and up to one month at - 30 degrees C without observing a change in the concentration of the other amino acids. No change occurred, when serum was stored at - 30 degrees C for 24 h.


[Measuring the activity of the placental isoenzyme of alkaline phosphatase: 3 methods compared (author's transl)]. Three methods were compared in human serum for determining the activity of the placental isoenzyme of alkaline phosphatase: 1. Heat inactivation at 65 degrees C for 10 minutes, 2. Differential inactivation with L-p-Bromotetramisole, and 3. an immunological precipation test. A good comparison between the three methods was found (correlation coefficient between 0.973 and 0.982). For long series or screening determinations for "Regan-isoenzyme" the differential inactivation with L-p-Bromotetramisole is preferred because of the short analysis time and the possibility of simple mechanisation.


Laboratory production of ergot alkaloids by species of balansia. Four species of Balansia (clavicipitaceous systemic grass pathogens) isolated from pastures where cattle showed signs of ergot toxicity were grown in culture. Balansia epichloë, one isolate of B. claviceps, B. henningsiana and two isolates of B. strangulans produced conidia in submerged culture during the first stage of a two-stage fermentation procedure. When tranferred to a glucose/sorbitol/inorganic salts medium during the second stage, these four species produced ergot alkaloids in stationary cultures. The transfer of fungi cultured in the first medium to the second medium was necessary for alkaloid biosynthesis. One isolate of B. claviceps did not produce alkaloids. Balansia epichloë produced chanoclavine (I), agroclavine, penniclavine, elymoclavine, ergonovine and ergonovinine. Balansia claviceps produced chanoclavine (I), ergonovine and ergonovinine. This is the first report of isolating ergonovine and ergonovinine, two lysergic acid derivatives, from fungi outside the genus Claviceps. Only chanoclavine (I) was identified from extracts of B. strangulans and B. henningsiana. Chanoclavine (I) and ergonovine were identified from smut grass (Sporobolus poiretii) parasitized by B. epichloë, indicating that this endophyte produces alkaloids both in vivo and in vitro.


Aflatoxins and anthraquinones from diploids of Aspergillus parasiticus. Three spore colour, two mycelial colour and 12 auxotrophic mutants were isolated from an aflatoxigenic strain of Aspergillus parasiticus. These mutants and heterozygous diploids formed by pairwise combinations of auxotrophs were assayed for aflatoxin production; norsolorinic acid and versicolorin A production were also assayed in the diploids. In general, introduction of an auxotrophic marker lowered aflatoxin production in haploids. The green-spored, prototrophic diploids resembled haploid wild-type strains in that they produced high levels of aflatoxin, low levels of versicolorin A, and no detectable norsolorinic acid. Parasexual analysis of segregants from four heterozygous diploids was hampered by the uniform conidiospore diameter of haploids and diploids and by the non-random recovery of genotypes among somatic segregants with and without treatment with p-fluorophenylalanine. Nevertheless, this technique is useful for recombinational analysis of mutants blocked in aflatoxin synthesis. The fortuitous association of mycelial pigmentation with certain blocked aflatoxin mutants should prove useful in future analyses of the genetics and biosynthesis of these economically important secondary metabolites.


Taxonomic significance of yeast sphaeroplast release after enzymatic treatment of intact cells. Treatment of whole yeast cells with a mixture of a reducing agent and 1,3-beta-glucanase isolated from Basidiomycete QM806 led to the production of sphaeroplasts from ascomycetes, from some fungi imperfecti, but not from basidiomycetes. Association of 1,3-beta-glucanase with a second enzyme, 1,4-alpha-glucanase, from Trichoderma viride, was required for sphaeroplast release from some, but not all, basidiomycetes and fungi imperfecti. The ability of yeast cells to liberate sphaeroplasts following appropriate enzymic treatment is proposed as a taxonomic criterion for differentiating basidiomycetous from ascomycetous yeasts and for classifying fungi imperfecti yeasts.