Local migration of myosin in F-actin plus ATP solution on the boundary of a diffusion cell. The diffusion phenomena of myosin (myosin A, H-meromyosin or subfragment-1) in F-actin plus ATP solutions were investigated. The upper part of the diffusion cell was filled with F-actin plus ATP, and the lower part was filled with F-actin, ATP, and myosin, then both parts were brought into contact so that a boundary of the two solutions was formed and the diffusion of myosin in F-actin plus ATP solutions started. The diffusion pattern was observed with a schlieren lens system. When almost all the ATP in the lower part of the cell had been consumed by actomyosin, a hyper-sharp schlieren pattern appeared near the boundary. On analyzing this pattern, it was found that a local fast migration of proteins was occurring. Simple Brownian motion of myosin molecules could not explain the hyper-sharp phenomenon. This phenomenon occurred in ther pesence of Mg2+ or Ca2+, but very little in the presence of EDTA. Although it is well known that the superprecipitation of myosin B suspension occurs only at physiological ionic strength, this phenomenon occurred over a relatively wide range of ionic strengths. The molecular mechanism of this phenomenon is discussed in relation to the basic mechanism of the interaction between myosin and F-actin.


Proteolysis of the major capsid protein T4 bacteriophage polyheads limited by quaternary structure. Bacteriophage T4 carrying an amber mutation in gene 22 plus an amber mutation in gene 21 form aberrant, tubular structures termed rough polyheads, instead of complete phage when they infect Escherichia coli B. These rough polyheads consist almost entirely of the major capsid protein in its uncleaved form (gp23). When rough polyheads are treated under mild conditions with any of the five proteases, trypsin, chymotrypsin, thermolysin, pronase, or the protease from Staphylococcus aureus V8, the gp23 is rapidly hydrolyzed at a limited number of peptide bonds. In contrast, cleaved capsid protein (gp23) in mature phage capsids is completely resistant to proteolysis under the same conditions. A major project in this laboratory requires determining the primary structure of gp23, a large protein (Mr = 58,000) quite rich in those amino acids at which cleavages are achieved by conventional means. Recovery of peptides from the complex mixtures resulting from such cleavages proved to be extremely difficult. The limited proteolysis of gp23 in rough polyheads had yielded a set of large, easily purified fragments which are greatly simplifying the task of determining the primary structure of this protein.


Phospholipid composition and membrane function in phosphatidylserine decarboxylase mutants of Escherichia coli. Temperature-sensitive conditional lethal mutants in phosphatidylserine decarboxylase (psd) accumulate large amounts of phosphatidylserine under nonpermissive conditions (42 degrees C) prior to cell death. In addition, the ratio of cardiolipin to phosphatidylglycerol is increased. At an intermediate temperature (37 degrees C), high levels of phosphatidylserine can be maintained with little effect on cell growth or viability. Under these conditions, both the rate of induction and the function of the lactose transport system are normal. At 42 degrees C addition of Mg2+ or Ca2+ to mutant cultures produces a partial phenotypic suppression. Growth is prolonged and the filaments normally present at 42 degrees C do not form. Upon transfer to the nonpermissive temperature, there is a considerable lag before accumulation of phosphatidylserine begins and the growth rate is affected. Based on the kinetics of heat inactivation of phosphatidylserine decarboxylase activity in extracts, in intact nongrowing cells, and in growing cells, it appears that the enzyme newly synthesized at 42 degrees C is more thermolabile in vivo than enzyme molecules previously inserted into the membrane at the lower temperature. Thus, the older, stable enzymatic activity must be diluted during growth before physiological effects are observed.


The peptide chain elongation factor genes tufA and fus of Escherichia coli are intimately related physically. Recent work from several laboratories has established the following points about the synthesis of the polypeptide chain elongation factors Tu and G in Escherichia coli. (i) Elongation factor Tu is the product of duplicate, highly conserved genes, tufA and tufB, which are widely separate parts of the chromosome. (ii) The molar  concentration of this factor is considerably higher than that of elongation factor G which is encoded by the fus gene. (iii) Although the tufA and fus genes are close together and can be co-transcribed in the direction from fus to tufA, the tufA gene product is synthesized at several times the rate of the fus gene product. In an attempt to understand what mechanism(s) could account for the differential expression of the tufA and fus genes, we sought to obtain more precise information on the physical relationship of these genes. By examining heteroduplexes between restriction endonuclease-generated fragments of DNA containing the tufA, fus, and tufB genes, we have demonstrated that the fus and tufA genes are intimately related physically in one of two possible arrangements. Either the NH2-terminal region of the tufA gene is contiguous with the COOH-terminal region of the fus gene or the beginning of the tufA gene overlaps part of the fus gene. These results mean that if the tufA gene is always co-transcribed with the fus gene, then some mechanism must allow the tufA portion of the transcript to be translated more often than the fus gene portion of the transcript.


Determination of the first half of the coat protein cistron of bacteriophage Qbeta as an application of a mapping procedure for RNA fragments. A method is described to classify, in regard to their location within the genome, fragments obtained by partial cleavage of 32P-labeled bacteriophage Qbeta RNA. The location of many fragments suitable for sequence analysis could be established using as markers 29 large RNase T1-resistant oligonucleotides with known map positions. Applying this method four fragments originating from the coat protein cistron were isolated and analyzed. The sequence of a segment of 239 nucleotides located immediately adjacent to the initiation triplet was determined to be G-C-A-A-A-A-U-U-A-G-A-G-A-C-U-G-U-U-A-C-U-U-U-A-G-G-U-A-A-C-A-U-C-G-G-G-A-A-A-G-A-U-G-G-A-A-A-A-C-A-A-A-C-U-C-U-G-G-U-C-C-U-C-A-A-U-C-C-G-C-G-U-G-G-G-G-U-A-A-A-U-C-C-C-A-C-U-A-A-C-G-G-C-G-U-U-G-C-C-U-C-G-C-U-U-U-C-A-C-A-A-G-C-G-G-G-U-G-C-A-G-U-U-C-C-U-G-C-G-C-U-G-G-A-G-A-A-G-C-G-U-G-U-U-A-C-C-G-U-U-U-C-G-G-U-A-U-C-U-C-A-G-C-C-U-U-C-U-C-G-C-A-A-U-C-G-U-A-A-G-A-A-C-U-A-C-A-A-G-G-U-C-C-A-G-G-U-U-A-A-G-A-U-C-C-A-G-A-A-C-C-C-G-A-C-C-G-C-U-U-G-C-A-C-U-G-C-A-A-A-C-G-G-U-U-C-U-U-Gp. The primary structure and the secondary structure model derived from it did not provide any evidence of homology with the corresponding RNA region of bacteriophage MS2.


Subunit conformation of yeast alcohol dehydrogenase. The primary structure of yeast alcohol dehydrogenase has been compared to the known tertiary structure of the corresponding horse liver enzyme after proper alignment of the two proteins. Possible influences on the subunit conformations of all amino acid exchanges, which affect 75% of the positions, were examined from interactions in the x-ray model of the horse enzyme. In spite of the differences, 90 of 93 strictly internal residues are similar, 18 space-restricted glycine residues are conserved, 16 structurally compensated exchanges occur, all functionally essential residues are similar or identical, and 41 gaps in either sequence may be accommodated in the model. These results show that the general subunit conformations and enzymatic mechanisms of the two enzymes are largely identical. Four surface areas are changed, affecting a region with differing charges, a noncommon loop, a structure around the second zinc atom, and residues at the main dimer interface. Although the subunit interactions in the yeast enzyme cannot be determined, the surface changes probably correlate with differences in quaternary structure between the proteins.


Differential scanning calorimetry of asparate transcarbamoylase and its isolate subunits. The thermal denaturation of aspartate transcarbamoylas of Escherichia coli was investigated by differential scanning calorimetry. Isolated regulatory and catalytic subunits were heat denatured at 55 and 80 degrees C, respectively. In contrast, the intact enzyme was denatured in two steps. A small endotherm near 73 degrees C was assoicated with denaturation of the regulatory subunits and the major endotherm at 82 degrees C with denaturation of the catalytic subunits. Thus regulatory subunits are stabilized against heat denaturation by more than 17 degrees C when incorporated in the enzyme. Similar conclusions were obtained from measurements of the enthalpy of heat denaturation. Regulatory subunits yielded a much lower value of the enthalpy of denaturation, 1.91 cal/g, than that found for the catalytic subunit, 3.94 cal/g, or typical globular proteins (4 to 6 cal/g). When the regulatory subunits were incorporated into aspartate transcarbamoylase their enthalpy of denaturation was increased 125% (to 4.3 cal/g). The enthalpy of the catalytic subunits in the intact enzyme was increased 38% (enthalpy of denaturation of 5.43 cal/g). Stabilization of the isolated catalytic subunit as well as the intact enzyme was achieved by the addition of the bisubstrate analog N-(phosphonacetyl)-L-aspartate. Similarly the allosteric effectors, CTP and ATP, stabilized the isolated regulatory subunits or those subunits within the intact enzyme. However, the addition of the bisubstrate analog caused a decrease in the enthalpy of denaturation of the regulatory subunits within the enzyme. These results are consistent with other studies of the ligand-promoted conformational changes in the native enzyme.


Spondylolysis. A skeletal investigation of two population groups. The lumbar spines of 485 skeletons of adult South African "Whites" and "Blacks" of both sexes were examined to determine the incidence and morphology of defects in the pars interarticularis. The overall incidence was 3.5%, without significant variation between races and sexes. The incidence of lumbar spina bifida in the whole sample was 1.9%, but was 11.8% in those skeletons with spondylolysis. Some instances of unilateral spondylolysis demonstrated obvious callus formation, suggesting a capability for normal repair. It is possible that the defects in bilateral cases represent established non-union of fractures of the pars interarticularis resulting from excessive mobility, and are not due to dysplasia of bone. It is noted, for the first time, that the superior facets of affected vertebrae are abnormally enlarged, and that the inferior facets of the separate neural arch are characteristically elliptical.


The site dependence of the articular cartilage transplant reaction. Allografts of immature joint cartilage from the knees of lambs were transferred heterotopically into an intramuscular site in animals which had been presensitised by two sets of skin grafts from the same donors. All of these grafts were found to be largely destroyed by the immune response as early as four weeks after transfer. Similar grafts transferred orthotopically into the knees of the recipients, on the other hand, were found to be thriving even after twelve weeks and evoked a minimal response. Heterotopic autografts also provoked a mild though non-specific inflammatory reaction which the orthotopic grafts did not. It is concluded that cartilage matrix is capable of protecting grafts to a remarkable degree even from a severe immunological assault but only when the nutrition is adequate. It is suggested that the conflicting results of similar previous experiments may be explained by variations in the nutritional state of the graft which may be affected by the technique of transplantation used.


Insulin binding sites on rat liver nuclear membranes: biochemical and immunofluorescent studies. Preliminary investigations (Horvat et al., '75) indicated the nucleus of rat liver as a site for specific binding of insulin. In this report these observations are confirmed. Nuclei from rat liver were isolated in a highly purified state as verified by interference contrast and electron microscopy and by chemical analysis. Extensive scanning of the preparations did not reveal the presence of structures resembling plasma membranes. The nuclear envelope was isolated by a modification of the method of Kay et al. ('72). Electron micrographs showed the presence of nuclear "ghosts" and few other recognizable nuclear elements, but no plasma membranes (60--80 A thick) were detected. The preparation was found to contain specific insulin binding activity. Specificity of the binding sites for insulin was demonstrated in competition studies with other polypeptide hormones and a synthetic insulin analog. Scatchard analysis of the binding data indicates the presence of a single class of high affinity receptors. In contrast to findings with plasma membranes the hormone-receptor complex is very stable and the kinetics of the dissociation of bound [125I]-insulin do not indicate negative cooperativity of the binding sites. Immunofluorescent labeling of intact, unfixed nuclei showed a specific fluorescent halo only around those nuclei that have been preincubated with insulin. All other controls were negative.


Specific inhibition of in vitro Candida-induced lymphocyte proliferation by polysaccharidic antigens present in the serum of patients with chronic mucocutaneous candidiasis. A specific inhibitory activity of in vitro proliferative responses of normal human lymphocytes to Candida metabolic antigen was found in the serum of 6 out of 23 children with chronic mucocutaneous candidiasis. In each of the six patients, the presence of an inhibitory activity was associated with Candida-specific cellular defects, characterized by a negative-skin test and a lack of in vitro lymphocyte proliferation. The presence of a circulating inhibitor was detected during relapses of the disease and disappeared under antifungal therapy. This inhibitory effect was not associated with any toxicity on tested lymphocytes. The factor was shown to be nondialysable, thermostable, nonprecipitable with ammonium sulfate and absorbable on anti-Candida antibodies or concanavalin A-coupled agarose columns. Altogether, these results suggest that the inhibitory factor is not an immunoglobulin, but rather a polysaccharidic antigen of Candida albicans. An inhibition of Candida-induced proliferative response of normal human lymphocytes was also obtained by addition of polysacharide antigens or purified mannans from C. albicans to cultures. Candida polysaccharidic antigens appeared, therefore, to be involved in specific depression of cellular functions observed in chronic candidiasis.


Reversal by methysergide of inhibition of insulin secretion by prostaglandin E in the dog. These studies were designed to examine whether interrelationships exist between serotonin and prostaglandin E (PGE) during regulation of insulin secretion in dogs in vivo. In our studies serotonin was found to inhibit insulin responses to intravenous glucose. This inhibition was not reversed by complete adrenergic blockade provided through combined phentolamine and propranolol pretreatment. This property of serotonin is similar to that of PGE which also inhibits glucose-induced insulin secretion in  vivo independently of adrenergic activity. To investigate whether these effects of serotonin and PGE are related, studies with methysergide (a serotonin antagonist) and indomethacin (a PGE synthesis inhibitor) were performed. Methysergide reversed the effects of both PGE and serotonin. In contrast, indomethacin did not diminish the inhibitory effect of serotonin upon insulin secretion. It is hypothesized that endogenous serotonin may play a role in the inhibitory effect of PGE upon insulin secretion in dogs in vivo.


Stimulation of endothelial cell prostacyclin production by thrombin, trypsin, and the ionophore A 23187. Prostacyclin (PGI(2)) is an unstable prostaglandin which inhibits platelet aggregation and serotonin release and causes vasodilation. The PGI(2) activity produced by monolayers of cultured human endothelial cells and fibroblasts was measured by the ability of their supernates to inhibit platelet aggregation in platelet-rich plasma, or to inhibit thrombin-induced [(14)C]serotonin release from aspirin-treated, washed platelet suspensions. Monolayers of cultured human endothelial cells, stimulated with sodium arachidonate, thrombin, the ionophore A 23187, or trypsin, secreted PGI(2) into the supernatant medium. Monolayers of fibroblasts produced PGI(2) activity only when stimulated by arachidonate. "Resting," intact monolayers did not produce detectable PGI(2), nor did monolayers treated with ADP or epinephrine. Production of PGI(2) activity was abolished by treatment of the monolayers with indomethacin, tranylcypromine, or 15-hydroperoxy arachidonic acid. The PGI(2) activity of the supernates was destroyed by boiling or acidification. Inhibition of thrombin with diisopropylfluoro-phosphate, and of trypsin with soybean trypsin inhibitor, abolished the stimulation of PGI(2) production by these enzymes. Production of thrombin at a site of vascular injury could, by stimulating PGI(2) synthesis by endothelial cells adjacent to the injured area, limit the number of platelets involved in the primary hemostatic response and help to localize thrombus formation.


Transient skin flora: their removal by cleansing or disinfection in relation to their mode of deposition. A suspension of Staphylococcus aureus deposited on the skin was much more effectively removed by soap-and-water washing when it had been spread and allowed to dry (mean survival 2%) than when it had been rubbed on to the skin (mean survival 29.9%); when antiseptics (70% ethyl alcohol, Hibiscrub without added water) were used, there was no difference between their action against bacteria dried on and bacteria rubbed on to the skin; both of these methods, and especially alcohol, were more effective than soap and water. When a detergent-disinfectant method (washing with Hibiscrub and water) was used, there was a significantly greater effect against rubbed-on than against dried-on bacteria; soap and water was slightly more effective than Hibiscrub and water against the latter. The need to reappraise methods of reducing transient skin flora in 'hygienic' hand cleansing and the tests used for this purpose are discussed.


Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections. A fluorescein isothiocyanate (FITC) technique and one based on peroxidase-antiperoxidase (PAP) were used to study the distribution of immunoglobulin (Ig) in cryostat and paraffin sections of human tonsil. Trypsin and other proteolytic enzymes were used to 'unmask' the antigen in paraffin sections. The effects of processing, and particularly of fixation, on the immunohistochemical response of tissues were studied. The FITC and PAP methods detected Ig in paraffin and cryostat sections equally well. The distribution of the antigen was the same with both methods but the PAP method was the more informative. Formaldehyde-sucrose solution proved more suitable for fixing tissues for immunohistochemistry than glutaraldehyde. Trypsin revealed antigen in parraffin sections more efficiently than pepsin, papain, or pronase. Surface Ig (s-Ig) could be demonstrated in trypsinised paraffin sections but less effectively than in cryostat sections. Trypsinised paraffin sections were, however, more suitable for intracellular Ig (c-Ig) than cryostat sections although the performance of cryostat sections could be improved by prior fixation with a coagulative fixative.


Assessment of a tissue transport-medium in preservation of tissue-fixed immunoglobulins and complement demonstrated by direct immunofluorescence. A pilot study with skin from SLE patients. The present work was undertaken in order to test the value of a tissue transport-medium (Histocon) for direct immunofluorescence studies. For this purpose one skin biopsy was performed on each forearm of 26 patients with systemic lupus erythematosus. One of the specimens was left in ice-cold Histocon solution for 4, 8, or 20 hours, and the other was immediately quick-frozen. The results of the immunofluorescence tests with the two methods yielded similar results. It is concluded that the solution allows the preservation of tissue-fixed immunoglobulins and complement during short periods of transport.


Long-term clinical effects, bioavailability, and kinetics of minoxidil in relation to renal function. Minoxidil was used to treat 26 patients (17 to 67 years old) with severe hypertension and varying degrees of renal function. Our object was to assess long-term clinical efficacy, kinetics (acute and chronic), and bioavailability of minoxidil in chronic renal insufficiency. Minoxidil, 27 to 30 mg per day, decreased systolic and diastolic blood pressure during the first three months of therapy. Between the third and 24th months (30 months in one patient) there was no further change. Propranolol or clonidine was needed to control heart rate, and furosemide or dialysis was needed to control edema induced by minoxidil. Renal function improved in some of the mildy azotemic patients. Minoxidil kinetics after the customary dose did not differ whether the drug was taken as tablet or solution. Kinetic parameters during chronic administration of minoxidil did not differ from those after acute administration. The kinetics in chronic renal insufficiency do not differ from these in subjects with normal renal function.


Insulin secretion in lactating cows: responses to glucose infused intravenously in normal, ketonemic, and starved animals. Changes in insulin concentrations of blood plasma were studied in lactating cows during and after a 1-h intravenous glucose infusion in 5 spontaneously ketonemic, 10 nonketonemic (normal), and 4 starved cows. A biphasic increase in insulin with a maximum 10 to 20 min after the start of the glucose infusion was followed by a sustained (1 to 2 h) elevation in the normal and ketonemic animals. In starved cows only the later phase was detectable. Insulin was higher in normal than in starved and ketonemic cows during both phases of the insulin secretion in spite of approximately identical glucose. Differences between starved and ketonemic cows were not major. Average rates of decline in sugar concentrations after the end of the infusion were higher in normal and ketonemic [half times 143 +/- 23 (standard deviation) and 166 +/- 22 min] than in starved cows (281 +/- 47 min). The low basal insulin of plasma and poor responses of insulin secretion following glucose infusions indicate sluggish beta-cell function in ketonemic cows.


Distribution of enterobacteriaceae in recycled manure bedding on California dairies. The distribution of coliform organisms on three California dairies which use a system of automatic flushing of alley ways and composted, recycled manure solids as free stall bedding for utilization of waste was studied. There was a wide distribution of Escherichia coli, Klebsiella, and Enterobacter and to less extent Citrobacter organisms. Klebsiella were not commonly in bovine feces. Composting manure solids effectively reduced coliform counts to few or to zero. However, if given proper conditions of moisture and temperature, coliforms could multiply to large numbers again in composted solids whether from survivors or from external contamination. Dried composted manure was satisfactory material for bedding of free stalls provided it was dried properly before application.


Micromorphologic features of cervical erosion after acid conditioning and its relation with composite resin. The dentin surface in cervical erosion appears relatively smooth with evidence of intratubular deposits. After acid conditioning, the tubules are made patent though some still contain relatively less soluble intratubular contents. Resin applied to acid-conditioned, eroded dentin penetrated approximately 30 micrometers compared to more than 100 micrometers into the tubules of similarly treated normal dentin.


Adhesive bonding of various materials to hard tooth tissues. XIV. Enamel mordant selection assisted by ESCA (XPS). Surface analysis by XPS (X-ray photoelection spectroscopy), also called ESCA (electron spectroscopy for chemical analysis), indicates that only certain cations are appreciably sorbed by enamel from an acid etching solution containing phosphoric acid and equimolar concentrations of candidate mordant salts.


Dental porcelain/Ni alloy interface reactions and their effective prevention. In a Be-bearing Ni alloy porcelainized without preoxidation, Be diffuses to the interface, oxidizes, and oxygen is supplied by the atmosphere at the porcelain surface and by reduction of SiO2 at the porcelain-alloy interface. On the alloy side of the interface, pits develop locally and near them Ni and Cr oxidize. In the oxide, Ni and Si are reduced to form nickel silicides. Preoxidation of the alloy prevents Be oxidation and Si reduction for several hours of porcelainization.


Properties of sealants containing bis-GMA and various diluents. Properties of commercial and a series of experimental pit and fissure sealants, based on Bis-GMA were determined using rapid test procedures. By proper selection of monomeric diluent and addition of a photocrosslinking agent, the characteristics of sealant formulations can be varied widely.


Synthesis of social surveys on noise annoyance. Since noise was first recognized as a serious environmental pollutant, a number of social surveys have been conducted in order to assess the magnitude of the problem and to develop suitable noise ratings, such that, from a measurement of certain physical characteristics of community noise, one could reliably predict the community's subjective response to the noise. Recently, the author has reviewed the data from social surveys concerning the noise of aircraft, street traffic, expressway traffic, and railroads. Going back to the original published data, the various survey noise ratings were translated to day-night average sound level, and an independent judgment was make, where choice was possible, as to which respondents should  be counted as "highly annoyed." The results of 11 of these surveys show a remarkable consistency. It is proposed that the average of these curves is the best currently available relationship for predicting community annoyance due to transportation noise of all kinds.


Comprehensive compilation of empirical ultrasonic properties of mammalian tissues. A detailed review of the literature on ultrasonic propagation properties of mammalian tissues and organs has revealed 144 papers containing compilable data. Over 1300 lines of parametric data are listed, including the tissue, species, age, specimen preparation, anatomical structure, pathology, temperature, measurement method, frequency, velocity, attenuation, acoustic impedance, and density.


Response to corticosteroids in chronic bronchitis. Corticosteroid drugs are often employed in the treatment of patients with chronic bronchitis. Although some patients respond favorably to such therapy, the characteristics of such patients are not known. Twenty-four patients with chronic bronchitis were treated with prednisone 30 mg daily or placebo for one week each in a double-blind crossover study. The following were monitored before and after each treatment period: physical examination, symptoms, peripheral blood eosinophil count, sputum cell exmination, forced vital capacity (FVC), before and after isoproterenol aerosol. Seven of 24 patients had an FEV1 increase greater than 30% of the control value on prednisone but not on placebo. Blood eosinophil count was elevated  (greater than or equal to 350/mm(3)) in 7 patients; 2 of these 7 improved on steroid. Sputum cell examination revealed preponderance of eosinophils in 1, and occasional clumps of eosinophils in 8. Seven of these 9 responded to steroid. Sputum but not blood eosinophilia is a good predictor of a favorable response to steroid therapy.


[The effects of heparin in underdevelopment of the fetus due to maternal vascular conditions (author's transl)]. The use of heparin to interfere with the appearance of fetal maldevelopment (or to correct it) is logical but the results are difficult to prove because it is difficult to be certain of early diagnosis at the time when therapy has the best chances of being efficacious. The authors compare two groups of pregnant women in whom the muscular vascular tone of the cervix is abolished. The second group was treated with heparin. The results are statistically significant and prove that treatment with heparin is well based in cases of underdevelopment of the fetus when it is due to abnormalities in the maternal microscopic circulation (acute atherosis). They confirm that the enzyme, hormone and ultrasound parameters return to normal from the beginning of treatment with heparin.


[Technique for bringing about haemostasis in surgery of extra-uterine pregnancy with conservation of the tube (author's transl)]. The authors describe a technique for haemostasis in the tubal ampulla after conservative surgery in ectopic pregnancy in the tube. This technique uses compression by a small balloon which is blown up inside the tube. It is placed there using a stilette which is introduced through the abdominal opening.


[The chemotherapy of advanced breast cancer. A report on the first 600 cases in a cooperative programme to carry out a systematic study (1974-1977) (author's transl)]. The authors explain the objectives and the structure of a programme evolved for the methodic study of advanced breast cancers which they have worked out and carried out in cooperation with the departments of medicine in eight French anticancer centres. They analyse the initial phases, which correspond to the three first years of work. They give the state of the results that have been obtained in 603 patients using three protocols for treatment. These consisted of the administration in interrupted doses of chemotherapy which consisted of vincristine associated for the one part with cyclophosphamide and 5-fluoro-uracil and on the other hand with doxorubicine or/and with methotrexate. From this study there emerged the value of combined therapy using doxorubicine as much because of the frequency with which results better than 50% were obtained (in a randomised trial) as by their quality, which was made clear in an important series of 240 cases. They point out the conditions required to initiate the place of such a scheme in all complex therapy which aims as much at palliation as at cure, and point out how important it is to be methodical in the application of chemotherapy in order to ensure its development as an anticancer therapy.


Quantation by flow microfluorometry of total cellular DNA in Acanthamoeba. The DNA content of five species of Acanthamoeba was determined by flow microfluorometry. Acanthamoeba castellanii (AC-30), acanthamoeba polyphaga (APG and P-23), acanthamoeba rhysodes, acanthamoeba culbertsoni (A-1), and acanthamoeba royreba were grown in a casitone based medium 24-48 HR. The trophozoites were harvested, and evaluated for DNA-bound fluorescence. All species tested has DNA values between 2.0-5.0 pg/cell. These results placed DNA/cell values of Acanthamoeba slightly lower than DNA/cell values of other eucaryotic cells and much lower than Amoeba proteus values. These results indicate that FMF may be a useful adjunct in distinguishing Acanthamoeba cells from either eucaryotic cells or some other amoeba. However, differences in DNA/cell between species of Acanthamoeba are small and would not be useful in identification of species.


Binding of deoxyribonuclease I to actin: a new way to visualize microfilament bundles in nonmuscle cells. Intracellular actin-containing fibers can be visualized by indirect immunofluorescence microscopy when they are stained with antibody directed against DNase I. The location of actin-containing fibers in cells appears to be similar to the staining pattern of antibody to actin. Actin fibers were also visualized by direct fluorescent microscopy with rhodamine-conjugated DNase I.


The accessory optic system and the retino-hypothalamic system. A review. The introduction of new techniques in neuroanatomy has helped to clarify a number of important issues on the nature of the accessory optic and retino-hypothalamic system. In this study, an effort was made, to collect all data obtainable, and to arrange them on the basis of some common parameter, so that the various results may be directly compared. These data were then interpreted on specific issues, although only limited coverage has been given to functional aspects, as described in older publications. The absence of a universally accepted concept of the function of both systems to date suggests the need for a thorough analysis of both contemporary and older literature dealing with a characterization of the structural components of the accessory optic and the retino-hypothalamic systems. On the basis of these anatomical studies it is concluded that the accessory optic system (AOS) is much more complex than formerly believed. Its development in various vertebrae classes is quite different. Among the mammals, the AOS is very well established in rodents but in primates and man its anatomical and functional role seems markedly reduced. In the rodents it consists of both superficially and deeply arranged axons which are in close topical relation to the cerebral peduncles. The axons not only supply the classic accessory otpic terminal nuclei but also the subthalamic nucleus and the substantia nigra. The secondary projections have not yet been established. The retino-hypothalamic system (RHS) mainly but not exclusively supplies the suprachismatic nucleus (sc). Crossed as well as uncrossed retinal fibres project to the ventro-lateral and to the posterior part of this nucleus. The cells within the sc combine both secretory and neuronal properties. The possible functions exerted are discussed. Both, the AOS and the RHS, are so closely related within the suprachiasmatic area that most methods fail in correlating experimental results with either system. The numerous problems which still remain to be clarified are also discussed.


Inhibition of the immune response by membrane-bound antibody. Cells from the spleens of "normal" swine, which were pretreated with pronase to remove surface membrane-bound immunoglobulin, gave an enhanced hemolytic plaque-forming cell response to sheep red blood cells in vitro in comparison with untreated controls. The enhancement could be abrogated by preincubating pronase-treated spleeen cells in preparations containing antibody to sheep red blood cells. This effect was demonstrated by autologous sera, immune sera, and all three known classes of porcine serum immunoglobulins, including IgM, IgA, and IgG and could be removed by absorption with sheep red blood cells. Surface membrane-bound antibody exerted its effect by binding to the nonadherent cell population. The response of normal spleen cells was unaffected by antibody treatment. Pronase-treatment was not mitogenic, did not function as a polyclonal B cell activator, and did not selectively eliminate T or B cells. The results indicate that removal of antibody from the surface of lymphoid cells enhanced the humoral immune response invitro and confirm that membrane-bound antibody can inhibit response to antigen.


Null cells in the mouse possess membrane immunoglobulins. Null cells lacking typical T and B cell surface markers were isolated from the spleens of congenitally athymic mice by using either nylon wool or Sepharose macrobeads conjugated with rabbit antibody to mouse IgM to remove B lymphocytes. Although these null cells were negative for surface immunoglobulin by the criterion of immunofluorescence by using rabbit antisera to mouse heavy or light immunoglobulin chains, surface immunoglobulins were readily demonstrable by two alternative and independent techniques. First, by using chicken antibody to the (Fab')2 fragment of mouse IgG, nearly all null cells were positive for immunofluorescence. Second, immunoglobulins could be detected in lysates of null cells radioiodinated by the lactoperoxidase-catalyzed reaction. Analysis of the surface immunoglobulins of null cells by radioimmunoassay and polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate suggested that they are 1) qualitatively similar to those of B cells and 2) present in amounts between 10 and 30% of those of B cells.


Immunologic suppression after oral administration of antigen. I. Specific suppressor cells formed in rat Peyer's patches after oral administration of sheep erythrocytes and their systemic migration. Rats given 10(10) sheep erythrocytes (SRBC) orally were found to contain specific suppressor cells to SRBC in their Peyer's patches (PP) and mesenteric lymph nodes (MLN) after 2 days of feeding. After 4 days of feeding, similar suppressor cells were found in the thymus and spleen, but they were missing in the PP or MLN. These suppressor cells effectively blocked IgM and IgG plaque-forming cell responses to SRBC in Mishell-Dutton cultures and delayed-type-hypersensitivity responses to SRBC when transferred to syngeneic recipients, but they did not affect responses to horse erythrocytes. The orally induced specific suppressor cells appeared to be T2 cells since their activity was eliminated by in vivo treatment of SRBC-fed rats with anti-rat lymphocyte serum but not by adult thymectomy. Because carrageenan partially relieved the suppression observed in culture, the actual suppressive mechanism may also involve a macrophage.


Demonstration of the in vitro phagocytosis of Treponema pallidum by rabbit peritoneal macrophages. Evidence has been provided for the in vitro phagocytosis of virulent Treponema pallidum by stimulant-induced peritoneal macrophages. After the 4-hr incubation of macrophages with T. pallidum, treponemal antigens associated with the macrophages are specifically stained using indirect immunofluorescent techniques. Phagocytized treponemes appear within the cytoplasm of macrophages as round, brightly fluorescent "bodies" observable in increasing numbers as the duration of the treponeme-phagocyte interaction increases. Their presence is significantly reduced in the cytoplasm of macrophages that have been treated with cytochalasin B, a known inhibitor of phagocytosis, and in nonphagocytic fibroblasts. Additionally, supportive evidence for T. pallidum phagocytosis in vitro has been provided by electron microscopic examination in which treponemes have been demonstrated within typical phagocytic vacuoles. This study also provides evidence that immune serum factor(s) significantly promote the phagocytosis of T. pallidum, although a contribution by heat-labile serum factors has not been demonstrated. The possible mechanisms of immune serum contribution and the implications of the demonstration of T. pallidum phagocytosis are discussed.


Rat skin main neutral protease: immunohistochemical localization. Specific antiserum against the purified rat skin main neutral protease was used in double layer immunofluorescent method to localize the enzyme in normal rat skin. The specific immunofluorescence was seen in dermal cells that were identified as mast cells on basis of their metachromatic granules. Enzyme histochemical staining with naphthol AS-D chloroacetate localized to the same cells that exhibited specific immunofluorescence. The granules of isolated rat mast cells also gave a specific reaction with the immunohistochemical technique. The results provide further evidence for the suggestion that rat skin main neutral protease is identical with the rat mast cell "chymase."


Filtration and immunoprecipitation in the elimination of DNA polymerase activity associated with bacterial contamination of sera positive for hepatitis B e antigen and its corresponding antibody. Samples of serum inoculated with Escherichia coli and serum that became contaminated with bacteria after exposure to a laboratory atmosphere demonstrated elevated DNA polymerase activity. The levels of activity were well within the range of values found in hepatitis B e antigen (HBeAg)-positive samples. The bacterial polymerase activity was markedly reduced by a single passage of serum samples through a 0.22-micron Millipore filter prior to analysis. Repeated filtration did not result in a substantial further decrease in polymerase activity. In sera that were heavily contamined with E. coli, however, filtration was not successful in reducing bacteria-associated polymerase activity to a base-line uncontaminated level. In such instances double antibody immunoprecipitation proved effective in elimination of bacterial activity. When bacterial contamination of serum samples is a possibility, specimens should be subjected to either Millipore filtration or immunoprecipitation prior to analysis, particularly when correlation of DNA polymerase activity with HBeAg and its corresponding antibody is attempted.


An outbreak in 1965 of severe respiratory illness caused by the Legionnaires' disease bacterium. In January 1977 an unsolved outbreak of infection at St. Elizabeth's Hospital (Washington, D.C.) that occurred in 1965 was linked with Legionnaires' disease. The link was made by fluorescent antibody testing with the bacterium isolated from tissues of persons with Legionnaires' disease in the 1976 outbreak in Philadelphia. In July and August 1965, an epidemic of severe respiratory disease characterized by abrupt onset of high fever, weakness, malaise, and nonproductive cough, frequently accompanied by radiographic evidence of pneumonia, affected at least 81 patients at St. Elizabeth's Hospital, a general psychiatric hospital. Fourteen (17%) of the affected patients died. Intensive epidemiologic and laboratory investigations in 1965 did not determine the etiology. The etiologic organism may have become airborne from sites of soil excavation.


Influence of pili on the virulence of Proteus mirabilis in experimental hematogenous pyelonephritis. The ability of heavily and lightly piliated Proteus mirabilis to infect the renal parenchyma was compared in a model of hematogenous pyelonephritis. Cortical abscesses occurred in 13 of 24 rats injected with lightly piliated P. mirabilis but in none of 24 rats challenged with heavily piliated organisms (P less than 0.001). Lightly and heavily piliated organisms were cleared from the vasculature equally rapidly and were also delivered to the kidney in equal numbers. During the first 24 hr, however, titers of the lightly piliated organisms in the kidney increased by 4 logs, whereas the heavily piliated P. mirabilis were virtually all eliminated. Pili are believed to mediate attachment to cell surfaces, and heavy piliation has been correlated with enhanced virulence when P. mirabilis invades the kidney across the pelvic mucosa. The results in this study suggest, however, that pili may adversely affect bacterial survival within the renal parenchyma.


In vitro studies of amphotericin B in combination with the imidazole antifungal compounds clotrimazole and miconazole. The clinically important polyene antibiotic amphotericin B, in combination with two antifungal imidazole compounds, clotrimazole and miconazole, was studied in vitro. With use of results of cytoplasmic leakage, metabolic heat output, and minimal inhibitory concentration studies, a definite antagonistic response was demonstrated. It is suggested that, if combined antifungal drug therapy is clinically indicated, the drug combination be tested against the isolate by the simple technique of measuring cytoplasmic leakage or by the more elaborate method of flow microcalorimetry.


Localization of an adenosine deaminase-binding protein in human kidney. A protein which specifically complexes with adenosine deaminase (complexing protein) has been localized in human kidney. Thin sections of renal tissue were treated with rabbit anti-complexing protein serum followed by fluorescein-labeled goat anti-rabbit gamma globulin serum. Complexing protein was detected in the proximal tubules of four normal kidneys. The glomeruli in sections from one of the four normal kidneys were also positive. Lung, liver, spleen, cardiac muscle, skeletal muscle, and kidney medulla were negative by this technique. The glomeruli as well as the proximal renal tubules of two patients with combined immunodeficiency disease-adenosine deaminase deficiency and three of seven patients with kidney disease contained easily detectable levels of complexing protein.


Reconstruction of the ossicular chain with an autologous bone cylinder, produced with a hollow drill. Technique and application: a preliminary report. An autologous bone cylinder is produced with a hollow drill. After modelling this bone cylinder, it is easy to reconstruct a stable ossicular chain in the middle ear.


Changing electrocardiographic recording technology and diagnostic accuracy of myocardial infarction criteria. Improved standards for evaluation of ECG measurement precision. An analytical study was performed to estimate the magnitude of the visual Q wave duration bias produced by pressurized ink round stylus electrocardiographic (ECG) recorders. With a paper speed of 25 mm/sec and the ECG tracing width of 0.25 mm, the visually measured Q waves are on the average 8 msec too short. The corresponding error with the older type flat stylus recorder is less than 2 msec. Considerable differences can thus be anticipated in the frequency of observed ECG abnormalities in studies which use different types of electrocardiographs. The effect of the visual Q wave duration measurement bias on the diagnostic ECG classification was investigated in a group of 237 patients with old myocardial infarction and 299 subjects with no clinical evidence of infarction. An 8 msec measurement bias toward too short Q wave duration was observed to result in a potential loss of diagnostic accuracy of about 25% in some ECG coding categories. As a corrective procedure, it is recommended that the baseline width produced by a round recording stylus of uniform thickness irrespective of the vertical deflection velocity should be less.


A terminating reagent for the peroxidase-labelled enzyme immunoassay. Addition of a low concentration of sodium azide completely stops colour development in a peroxidase-labelled enzyme immunoassay using 2,2'-azino-di(3-ethylbenzthiazoline sulphonic acid-6) as substrate. Whereas high concentrations of azide cause a decrease in the colour intensity, the low concentrations used here have no significant affect on the colour.


Tissue cultures from cerebrospinal fluid specimens in the study of human brain tumors. The authors report a study in which 109 cerebrospinal fluid (CSF) specimens from patients with varying neurological disorders were incubated in tissue culture medium for 1, 3, and sometimes 7 days. Strict criteria for malignancy were applied to cells found at these intervals. In 35 patients with verified central nervous system neoplasms, eight cases had malignant cells and 11 others had "doubtful" cells by tissue-culture analysis. Thirty-three of these cases were also examined with standard millipore cytological techniques: six had malignant cells and four had "doubtful" cells. Of 50 cases with inflammatory or other non-neoplastic conditions, cells were cultured in 13. None was considered malignant by our criteria. Tissue culture of CSF has several potential benefits. Even with stringent criteria, it is possible to demonstrate the presence of unequivocally malignant cells in CSF by tissue culture. The systemic application of such criteria may eventually increase the positive identification of malignancies. Further, since these cells are growing, the degree of malignancy may be more accurately determined by a study of growth in culture. Such a study could not be done by conventional methods. Finally, tissue culture can help to guide therapy in certain instances in which a surgical biopsy cannot be obtained.


Experimental dysraphism in the rat.

Experimental dysraphism in the rat. An experimental treatment that results in dysraphic lesions in rat embryos was studied to gain insight into the origin of such lesions. Pregnant rats given 100,000 units of vitamin A by gavage tube on gestational Days 8, 9, and 10 produced abnormal litters, and 50% of the embryos had exencephaly and/or spina bifida. Study of these embryonic lesions revealed hyperplesia of neural tube tissue that extended laterally to the edge of an epidermal defect. The epidermal defect consisted of the absence of fetal epidermis over the hyperplastic neural tissue. At no stage in development was embryonic hydrocephalus noted. In fact, the central canals of the embryos with malformations were poorly developed and smaller than those of control embryos. Results with this model indicate that the malformations arise through a process of overgrowth and hyperplasia and not from embryonic hydrocephalus. Continued work with this and other models is required to delineate new mechanisms and to offer alternative postulates for the origin of dysraphic lesions.


Pyridoxine and its relation to lipids. Studies with pyridoxineless mutants of Aspergillus nidulans. The effect of pyridoxine deficiency on fat metabolism was studied using mutant strains of Aspergillus nidulans requiring pyridoxine for growth. Under pyridoxine deficiency the mutants exhibited increased levels of total lipid, sterols, phospholipids, and triacylglycerols. Total fatty acids were found to decrease with pyridoxine deficiency. An increase in saturated fatty acids and decrease in unsaturated fatty acids were seen with deficiency. Pyridoxine deficiency also increased lower carbon chain fatty acids. A possible involvement of pyridoxine in the elongation of fatty acid chain and in the desaturation of fatty acids in Aspergillus nidulans is suggested.


Long term preventive aspects in oral rehabilitation of adults and elderly. I. Maintenance of balanced functional jaw interaction. Prevention should be regarded as more important for the health than the cure. In adult and geriatric patients prevention is directed towards limiting disability rather than preventing the onset of the disease, since treatment is undertaken after pathological changes have taken place and some degree of invalidity is already present. Oral rehabilitation, if successfully performed, will limit the spread and further deterioration of oral conditions at any developmental stage of a functional pathology. The objective of the rehabilitation should not be the integrity of a single dental arch and certainly not the replacement of the lost teeth. Rather, the principle guiding guiding the treatment should be the achievement of a harmonious functional relation between the maxillary and mandibular jaws, so as to enable them to function in well balanced co-operation and force distribution. This objective may be best achieved by sensible planning and faultless execution, when the patient is younger, and most of his teeth and their supporting structures are still preserved.


The in vitro fungicidal properties of Visco-gel, alone and combined with nystatin and amphotericin B. The in vitro effect of the tissue conditioner Visco-gel, with or without the inclusion of nystatin and amphotericin B, upon Candida albicans, Candida krusei and Candida tropicalis, has been studied. Visco-gel alone was completely inert and would therefore not be used without nystatin in the treatment of a denture stomatitis where a yeast infection was present. Amphotericin B became completely ineffective when mixed with Visco-gel, but in control tests remained very active.


The adaptation of composite materials to etched enamel surfaces. The adaptation of three composite materials and one unfilled bis-GMA resin to etched enamel surfaces was investigated by examining under optical and scanning electron microscopy the adhesive-enamel junction of replicas. The length of the tags and the density of tagging observed on the replicas showed that the three composite materials, Adaptic, Concise and Delphic, adapted to the etched enamel surface equally as well as the unfilled resin Nuva Seal. The results of this study together with available results of mechanical testing and clinical studies allow the conclusion that the successful retention of the composite filling materials to an etched enamel surface is not dependent on the use of an intermediate unfilled resin or bonding agent.


The influence of surface roughness on the retentive ability of two dental luting cements. Two series of brass cones and two series of dentine posts with varying surface roughness were produced. Maximum roughness value and arithmetical mean roughness were recorded for each cone. Brass crowns were cemented either with zinc phosphate cement (De Trey's zinc Cenment Improved) or with polycarboxylate cement (Durelon). A tensile stress was applied until the crown and the cone separated. The retentive force in relation to retention area was measured. The results showed that the retentive ability of both cements increased with increasing surface roughness. The increase in retention was greater for brass than for dentine. Differences were also found between the two cements; on smooth surfaces the zinc phosphate cement had a lower retentive ability than the polycarboxylate cement, but on rough surfaces the opposite relationship was observed.


Review of bone grafting for reconstruction of discontinuity defects of the mandible. Thirty-eight patients with discontinuity defects of the mandible were hospitalized at Iwate Medical University School of Dentistry during the 12-year period from 1965 to 1976. Treatment consisted of grafting autogenous ilium and rib in 35 patients and isologous ilium and Kiel bone in three patients. No bony union was noted in four of 28 cases involving benign tumors, three of eight cases involving malignant tumors, and one of two cases involving old fractures. Postoperative infection was the main cause for lack of union.


Total maxillary advancement with and without bone grafting. Computer morphometrics was used to analyze 21 adult patients who underwent total maxillary advancement; ten had bone grafting behind the tuberosity and 11 did not. Each group was subclassified as those with idiopathic maxillary deficiency and those with cleft lip and cleft palate; all patients were treated by the same basic operative technique. The results of the study indicate that those patients who had bone grafts had optimum stability. However, individuals with maxillary deficiency who undergo minimal advancements (less than 0.5 mm) can have overcorrections at surgery and, therefore, do not require bone grafts for good stability.


Overwhelming postoperative streptococcal infection. Fulminant postoperative group A beta-hemolytic streptococcal infections present a serious and potentially lethal threat to children. Three children with severe postoperative streptococcal wound infections are presented. Each child developed high fever within the first 24 hr after surgery, followed shortly by signs of septic shock. In all patients, the clinical appreciation of wound infection became apparent late in the course of the illness. All three children had white blood cell counts below 6000/cu mm. One child developed crepitus in the incision; another showed radiographic evidence of free air in the soft tissues. The only survivor received antibiotics immediately following the initial fever along with re-exploration and drainage of the operative wound.


Modification of pulmonary vascular responses to arachidonic acid by alterations in physiologic state. The effects of bolus injections of arachidonic acid and prostaglandins (PG) E2 and F2a on the pulmonary vascular bed were compared under resting conditions and after alteration in the physiologic state of the lung. Studies were carried out in the vascularly isolated lung lobe of the intact, anesthetized dog under conditions of controlled blood flow. Arachidonic acid, PGE2 and PGF2a increased pulmonary vascular resistance by constricting intrapulmonary veins and arteries in a dose-related manner, as did an analog of the endoperoxide, PGH2, whereas PGI2 dilated the pulmonary vascular bed. The response to arachidonate was associated with a 2- to 3-fold increase in levels of PGE- and PGF-like substances in pulmonary venous blood and was blocked by indomethacin. The effects of arachidonic acid, but not the PGs, were greatly enhanced during perfusion with either dextran or saline and the enhanced response in saline was associated with a 15 to 20-fold increase in levels of PG-like substances in the pulmonary effluent. Responses to arachidonate were not dependent upon the presence of formed elements in blood but were related to perfusate protein concentration. Alveolar hypoxia decreased responses to the precursor while those to PGE2 and PGF2a were enhanced. Responses to the PGs, but not those to arachidonate, were affected by changes in blood pH. Sublethal doses of Escherichic coli endotoxin increased the response to arachidonic acid, but not those to PGE2 and PGF2a. Results of the present study indicate that the effects of bolus injection of arachidonic acid on the pulmonary vascular bed are due mainly to formation of constrictor metabolites which may overshadow the actions of any dilator (PGI2) formed and suggest that metabolism of the precursor is altered by changes in physiologic state.


Plasma LH concentrations after single or double injections of synthetic LH-RH in dairy cows. Milked Friesian dairy cows were given 1 or 2 i.v. injections of synthetic LH-RH. The second of 2 injections of LH-RH given 1.5 h apart induced a significantly greater release of LH than did the first. The response to this injection was also significantly greater than that caused by injections given 6 h apart. There was no significant difference between the LH responses to the two injections given 6 h apart. The total LH release produced by the two injections of 100 microgram synthetic LH-RH given 1.5 h apart was significantly greater than that produced by single injection of 200 microgram synthetic LH-RH. It is concluded that there is a short-term increase in the responsiveness of the bovine pituitary gland to LH-RH after an initial LH-RH stimulus.


Effect of removal of the zona pellucida on subsequent development of mouse blastocysts in vitro. Removal of the zona pellucida allowed mouse blastocysts incapable of hatching in vitro to develop in culture. Blastocysts denuded by pronase always developed further than those of identical age that had hatched spontaneously. More blastocysts mechanically denuded and treated with pronase developed egg cylinder-like structures than did blastocysts mechanically denuded and not treated with pronase. Plastic support gave better development of blastocysts than did glass.


Non-specific effects of passive immunization on implantation in the rabbit. Passive immunization with goat anti-rabbit uteroglobin antiserum prevents implantation in the rabbit. The dose of antiserum was too low to neutralize all of the uteroglobin present on Day 5 of pregnancy, however, and the effect could not be shown to be specific, because 'control' treatments with goat antiserum to chick avidin or normal goat serum also prevented implantation. Non-specific antisera raised in rabbits had little or no effect on implantation. Partial purification of antibodies from the non-specific goat antisera reversed their effect, while anti-uteroglobin gamma globulin still reduced implantation. Fluorescein-labelled gamma globulin fractions of anti-avidin and anti-uteroglobin both bound to blastocysts, but pure FITC-IgG showed binding only of anti-uteroglobin. Both anti-avidin and anti-uteroglobin IgG prevented implantation. It is concluded that the effect on implantation is not necessarily achieved via a specific antigen.


Evidence of a uteroglobin-like protein in epithelial cells of reproductive and non-reproductive tissues of the rabbit. An antiserum raised in a goat to a uteroglobin-like protein isolated from uterine fluid of oestrous rabbits was used in an immune fluorescence test to localize an antigen present in the reproductive tract of oestrous and pseudopregnant rabbits and mammary gland tissue. The antigen was also present in the vas deferens and seminal vesicle, but not in testis. Non-reproductive tissues, such as lung, small intestine, bladder and thyroid showed specific fluorescent staining which was eliminated or significantly reduced by absorption of the antiserum with a purified uteroglobin preparation.


Endotoxin-induced interruption of early pregnancy in the rabbit. Endotoxin derived from Salmonella enteritidis (Boivin) completely interrupted pregnancy in the rabbit when given as a single dose (10 or 20 microgram/kg) intravenously on Day 4 or Day 8 of pregnancy. Indomethacin (2 or 10 mg/kg) was unable to prevent this effect.


A histochemical study on the effects of photoperiod on gonadal and adrenal function in the male bank vole (Clethrionomys glareolus). NADH- and NADPH-diaphorases, 3alpha-, delta5-3beta-, 11beta- and 17beta-hydroxy-steroid dehydrogenases (HSD) and lipids were studied histochemically in the testes and adrenals of male bank voles kept in a long (16L:8D) or a short (8L:16D) photoperiod (Groups L and S, respectively). At 67 days of age the Group L males were heavier and had active and significantly larger testes than Group S males. The testes of Group S males were regressed and were also significantly smaller than those of 18-day-old animals born and reared in a 18L:6D photoperiod. Lipid droplets were detected in the Leydig cells and intratubular spaces in the testes of Group L animals, but were absent from those of Group S voles. The adrenal cortex of the Group L animals was virtually devoid of lipids, but large lipid inclusions were present in the basal zona fasciculata of the Group S voles. In the Group L testes the diaphorase activities were more intense and the difference in enzymic activity between the seminiferous epithelium and the Leydig cells was more pronounced (especially for NADH-diaphorase) than that in the testes of Group S animals. Moreover, the 3alpha-- and delta5-3beta-HSD activities were much stronger in the testes of sexually active animals; 17beta-HSD activity was present in the Leydig cells of the active testes, and absent in the regressed testes. There was no marked difference between the two groups of animals with regard to the distribution or intensity of diaphorases, 3alpha-, delta5-3beta-, 11beta- or 17beta-HSD in the adrenal cortex. It is concluded that a decline in steroid synthesis occurs in the testes of voles kept in a short photoperiod. The large lipid inclusions observed in the adrenal cortex of such animals suggest decreased corticosteroid synthesis and/or secretion.


Medical service plans in academic medical centers. Medical service plans are of major importance to academic medical centers and are becoming increasingly so each year as evidenced by growing dependence of medical schools on resulting funds. How these funds are generated and used varies among schools. The procedures may affect the governance of the institution, modifying the authority of the central administration or the clinical departments. Recent developments in federal legislation, such as health maintenance organizations and amendments (Section 227) to the Social Security Act, and the future development of national health insurance will certainly have an effect on how academic medical centers organize their clinical activities. How successfully various medical schools deal with the dynamic problem may well determine their future survival.


Application of statistical methods in quantitative microscopy. The purpose of the paper is twofold: first, to describe the aspects of quantitative microscopy where statistical ideas are being applied today; and second, to describe some ways in which more complete use of statistics could improve quantitative microscopy. The typical estimation problem in quantitative microscopy is described with emphasis on the modelling of variability in estimation. A Poisson field model for imbedded particulates is used to illustrate the value of theoretical treatments of variability. Sampling, estimation and multivariate analysis are cited as areas of statistics presently used by quantitative microscopists. The potential role of statistics in quantitative microscopy is discussed. Examples from particle sizing and characterization of alveolar lung structure illustrate the value of statistical ideas. Automatic image analysers, while making the collection of large amounts of data feasible, have complicated statistical estimation. An investigation of the size distribution of random chord lengths through alveolar chambers illustrates how statistical methodology must be modified for valid inference.


Special specimen preparation methods for image processing in transmission electron microscopy: a review. One of the important developments in quantitative electron microscopy has been the application of optical and computer imaging methods to electron micrographs. In general these techniques of image analysis have been applied to electron micrographs from isolated biological structures prepared in the presence of various negative stains. To make full use of image processing techniques there are obvious advantages in preparing suitable specimens containing large areas of repeating features. However, the number of naturally occurring biological specimens exhibiting crystalline or paracrystalline features suitable for high resolution electron microscopy and subsequent image analysis is relatively small.Some recent experiments on the in vitro formation of crystalline and paracrystalline arrays from highly concentrated and purified isometric, filamentous and rod-like viruses is reviewed. The problems associated with the preparative procedures for producing two-dimensional and three-dimensional crystalline arrays are discussed together with the possibility of extending the negative staining-carbon film method for studying the gradual dissociation or assembly of viral components.


Critical point drying for scanning electron microscopy: a semi-automatic method of preparing biological specimens. Slow, controlled, rates of critical point bomb heating and of gas venting have been shown to improve the preservation of biological specimens in critical point drying. A procedure that represents a balance between avoidance of specimen damage and speed of operation has been developed for use with CO2 as the transitional fluid. Bomb heating is automated and controlled electronically, and manual venting of the gaseous CO2 is monitored using a gas flow meter.


Extraction of glycoproteins during tissue preparation for electron microscopy. The extraction of glycoproteins labelled with 3H or 14C precursors: fucose and glucosamine, has been compared during processing of rat liver for transmission electron microscopy. Using 14C labelled fucose, approximately 6% of labelled macromolecules were extracted during processing including 3% during glutaraldehyde fixation, 1% during post-fixation and 2% during dehydration. Greater extraction (8%) occurs with glucosamine as a precursor, which may be attributed to hydrolysis of the more labile sialic acid residues of the glycoprotein molecules. Tritium labelled glycoproteins were more susceptible to extraction than 14C-labelled glycoproteins. The extraction of 3H-labelled glycoproteins as measured by liquid scintillation counting may prove difficult to interpret owing to the quenching from processing solutions.


A simple method for maintaining relative positions of separate tissue elements during processing for electron microscopy. Molten (328 K) 20% gelatin is used as a 'glue' to hold together separate tissue elements or tissue elements that may be separated when cutting small blocks of tissue for plastic embedding. Standard aldehyde and osmium fixation, dehydration and epoxy embedding are compatible with this as is semi-thin sectioning for light microscopy or thin sectioning for electron microscopy.


Estimation of the mean caliper diameter of cell nuclei. I. Serial section reconstriction method and endothelial nuclei from human lung. In morphometric studies of lung tissue, accurate determination of the total number of any specific type of cell, such as the endothelial cell, requires knowledge of the shape of the cell nucleus. This knowledge of shape is necessary to calculate the mean caliper diameter, which, in turn, is an indispensible element of the equation for the number of nuclei (therefore cells) per unit volume (NV). Nine human hung endothelial cell nuclei were therfore reconstructed in dental wax from serial sections and were found to be pleomorphic triaxial ellipsoids. Five of these approached an oblate shape. The axes of these nine nuclear models were directly measured and a computer program using numerical integration was written to determine the mean caliper diameters of these ellipsoids. The estimated mean semi-axes of the nine nuclei were (+/-SD) 5.98+/-1.61, 3.61+/-0.70, 1.36+/-0.34; and the estimated overall mean caliper diameter for the total population of human lung endothelial nuclei was 7.97+/-1.27 micrometers.


Estimation of the mean caliper diameter of cell nuclei. II. Various cell types in rat lung. Morphometric studies of lung tissue in which the number of cell nuclei (therefore cells) are determined require the determination of the mean caliper diameter for each type of cell nucleus studied. We determined the mean caliper diameter for the five most common classes of cells found in the gas exchange portion of rat lung. This was done by serially sectioning through ten nuclei from each class of cells and then constructing wax models of the nuclei. Most of the nuclei were found to be triaxial ellipsoids and direct measurements taken from the models were used to calculate the following estimated overall mean caliper diameters (+/-SE): alveolar type I nuclei, 7.97+/-0.2 micrometers, alveolar type II nuceli, 6.77 +/-0.19 micrometers, interstitial cell nuclei, 7.64+/-0.35 micrometers, endothelial cell nuclei, 7.22+/-0.19 micrometers, and alveolar macrophage nuclei, 7.65+/-0.17 micrometers.


Area changes in slices of rat brain during preparation for histology or electron microscopy. Cerebral slices cut from rat brain, either 2-3 mm or 0.27 mm thick, were used to study the effect of embedding and freezing. Paraffin wax sections 6 micrometer thick were mounted and stained with haematoxylin and eosin or Marsland et al.'s (1954) silver stain, and their areas were examined at each step. Embedding in paraffin wax of slices 2-3 mm thick, or in Epon of slices 0.27 mm thick, caused a diminution of their areas by 20-30%. Staining of paraffin wax sections did not alter their areas. Glycerol alone at 15% concentration had no effect on the areas, but at 30% concentration they were diminished by approximately 20%. Diminution of the areas of glycerol treated slices 0.27 mm thick also occurred when they were transferred to liquid N2 or to isopentane, but the areas increased after glycerol was replaced by Freon 12. It was concluded that embedding or freezing cerebral slices caused changes in their areas, but that staining of sections after they had been embedded, sectioned and mounted did not.


The use of simple image analysers in lung morphometry. We have used several commercially available image analysers to measure the inter-alveolar wall distance on histologic sections of animal and human lungs. Inter- and intra-observer variations using these machines are small, and very similar results are obtained using different machines. The recorded result depends on the magnification used since higher resolution produces more intercepts. The plane in which the measurement is made affects the result, representing the difference in shrinkage from fixed tissue to stained slides in the horizontal and vertical planes. Machine-measured inter-alveolar wall distances are smaller than those measured by humans because of the spurious intercepts made by the image analysers. However, the measurements are highly correlated and human-measured inter-alveolar wall distances can be closely predicted from the machine-measured ones. The number of alveoli per unit volume is less well predicted from the average inter-alveolar wall distance. Image analysers may possibly play a useful role in making similar simple measurements in other organs.


A new microwave device for rapid thermal fixation of the murine brain. A microwave applicator was developed to provide rapid thermal fixation of enzymes in the murine brain. Although the power output of the device is only 1.3 kW, whole-brain levels of acetyicholine after microwave fixation are comparable to those observed at a power output of 5 kW, and are markedly higher than those obtained by a conventional method (freezing) of fixation.


Effect of unblocking therapy and levamisole on primary gastrointestinal tumors in rats: immunologic and histologic correlation. The effect of multimodal immunotherapy was studied in WF rats bearing primary gastrointestinal (GI) tumors induced by 1,2-dimethylhydrazine dihydrochloride. The alterations induced in antitumor immune responses of the treated rats were studied in vitro and were correlated with tumor status in vivo. Multimodal immunotherapy consisted of unblocking serum, unblocked lymphoid cells, and levamisole. Such immunologic intervention resulted in significant inhibition of tumor growth, inhibition of metastases, and prolonged survival of the host. Serum blocking activity could be completely counteracted in 6 rats, all of which showed complete tumor regression. Of 20 rats, 8 showed inadequate counteraction of serum blocking activity and transient appearance of cytotoxic antibodies. All 8 rats showed marked tumor inhibition and prolonged survival. Six remaining rats succumbed from either GI or extra-GI tumors, although they survived significantly longer than untreated rats; these 6 rats had only transient counteraction of their serum blocking activity. All 20 tumors in 14 rats of the therapy group showed histologic evidence of tumor rejection. Our studies suggested that a complete counteraction of blocking activity in conjunction with methods capable of improving the specific and nonspecific immune competence of the host may be important to achieve optimal antitumor effects.


The use of pedicled grafts of omentum in the repair of transplant-related urinary tract problems. Urologic complications related to renal transplantation may be difficult to treat and can be serious threats to life or to function of the transplanted kidney. Successful experience with omentrum for non-transplant and transplant-related reconstructive procedures recommends its further use. Herein are reported 4 patients in whom pedicled flaps of omentum were used to treat transplant-related urologic problems.


Autosuture ileal conduit construction: experience in 110 cases. Our experience with 110 cases of ileal conduit construction with autosuture stapling devices is discussed. The technique is described briefly and the results are compared to results with ileal conduits constructed with the conventional suture technique. Analysis of our data revealed a significant reduction in operating time and postoperative morbidity.


Prazosin and clonidine for moderately severe hypertension. In a single-blind comparative study of the cases of 30 moderately hypertensive patients, clonidine hydrochloride and prazosin hydrochloride had similar effectiveness in lowering blood pressure. Neither agent had significant effects on the renin-aldosterone axis. Addition of polythiazide to prazosin and chlorthalidone to clonidine notably increased the antihypertensive effect of both drugs. Serum cholesterol levels were observed to decrease when prazosin and clonidine were given and to rise when the diuretics were added to the regimen. The patients treated with clonidine were troubled by side effects, particularly drowsiness and dry mouth. Prazosin was better tolerated, with side effects tending to diminish with time. The "first-dose" effect was seen in two patients given prazosin, but it did not limit treatment. Both diuretics induced notable hypokalemia.


Chorio-meningo-encephalitis due to Listeria monocytogenes after renal transplantation. A 35-year-old man, who underwent renal transplantation, died of meningo-encephalitis with symptoms of meningitis for only 8 days. The autopsy revealed marked chorio-meningo-encephalitis caused by Listeria monocytogenes. This report on listeriosis in renal transplant patients is the second in our country, where adult cases of listeriosis are rare. Recent knowledge about listeriosis in renal transplants is reviewed and discussed.


Gangliosides and thermal adaptation in vertebrates. Gangliosides, which are highly enriched in synaptic membranes, show great differences in concentration and pattern constellation as well during early ontogenetical development as on interspecies level in vertebrates. As, up to now, there is no reasonable explanation for these findings, and as it is assumed the synapse to be the primary site of thermal adaptation, the attempt was made to investigate whether there are any correlations between brain gangliosides and the thermal adaptation phenomenon. 1. While in the brains of adult homeothermic vertebrates (with thermo-regulation: mammals, birds) the di-sialoganglioside GD1a predominates, in the brain of poikilotherms (without thermo-regulation: e.g. amphibia, teleost fishes) more polar polysialogangliosides are present. 2. In homeotherms during their early perinatal phase (heterothermic phase: thermor-regulation being not yet developed) a temporary poly-sialisation of brain-gangliosides occurs. 3. In poikilotherms, during the process of thermal adaptation to lowered environmental temperatures, a poly-sialisation of brain gangliosides can be observed, as well during the phase of acclimatization (adaptation to seasonal changes in temperature) as also to acclimation (experimentally induced changes in the environmental temperature). 4. The phenomenon of poly-sialisation of brain gangliosides during adaptation to lowered environmental temperatures can be correlated with changes in some behavioral (e.g. motorical activity) and electrophysiological parameters. 5. On the background of a general hypothesis on the involvement of gangliosides in the process of transmission [23, 24], a functional model on the participation of gangliosides in the process of thermal adaptation is discussed with special regard to the formation of Ca++-ganglioside-complexes, which are highly sensitive to temperature changes.


Mode of immunopotentiating action of BCG: persistence and spread of BCG infection. The mode of action of BCG, strain Japan was investigated using the immune response to sheep red blood cells (SRBC) as an indicator system. When SRBC were injected into the BCG-primed foot pad, the direct plaque forming cells (PFC) and the effector cells responsible for delayed type hypersensitivity (PTH) were produced in various lymphoid organs widely distributed. Furthermore, bacterial counts in the draining popliteal lymph node and the spleen in the mice inoculated with BCG into the hind foot pad suggested that a local infection with BCG has spread to a generalized systemic infection of lymphoid tissues with time. Enhancement of DTH response to SRBC was induced when the mice previously infected with BCG were inmunized with SRBC mixed with purified protein derivative (PPD). These findings suggested that nonspecific augmentation of immune response with BCG was due to a generalized systemic activation of lymphoid system by BCG infection and the long lasting effect of immunopotentiation with BCG was due to persisting BCG infection in the lymphoid tissues.


Experimental Pseudomonas infection in mice: acquired resistance against Pseudomonas septicemia and altered susceptibility in BCG infected mice. Pseudomonas septicemia in mice caused the early death in the first three days of infection without any localized lesions. Localized lesions such as abscess are observed in the kidney and liver after the third day of infection. The early death increased in the BCG infected mice showing an increased level of macrophage activation. It seemed that such early death is due to endotoxin produced by Pseudomonas aeruginosa. However, the BCG infected mice showing an enhanced antibody formation were more resistant to pseudomonas septicemia. Immune serum protected such death, but immune spleen cells did not. Furthermore immune serum also protected the increased death in BCG infected mice.


Drug resistance of Escherichia coli isolated in Nepal. We collected Escherichia coli strains from 59 Nepalese porters in 1971 and surveyed for their drug resistance. Drug-resistant E. coli strains were isolated from four porters. (TC. CM. SM. SA. APC.)-resistant strains were isolated from two porters and SA- or APC-resistant strains were isolated from each of the others. The R factors were demonstrated from the multiple-resistant E. coli strains.


[Method of surgical treatment of aneurysms of the peripheral vessels]. The author advocates active surgical tactics in aneurysm both in the stage of pulsatile hematoma and in immature aneurysm since only operation may prevent the development of such complications as hemorrhage, suppuration of the aneurysm, thrombosis of a major vessel, bacterial endarteritis, and bacterial endocarditis. The development of specialized medical service makes it possible to conduct a skilled operation, which greatly reduces the term of the patient's disability.


Analgesic nephropathy: etiology, clinical syndrome, and clinicopathologic correlations in Australia. Analgesic abuse is a major public health hazard in Australia, and analgesic nephropathy with consequent terminal renal failure is the underlying cause in 20% of the patients requiring dialysis and transplantation. Analgesics are invariably taken in the form of compounds and mixtures. In the aspirin-phenacetin-caffeine (APC) mixture, aspirin appears to be the major nephrotoxic agent and phenacetin appears to play a secondary and synergistic role. The renal disease associated with abuse of analgesics is characteristic and is part of a much wider clinical syndrome, the analgesic syndrome, which includes peptic ulcer disease (35%), anemia (60 to 90%), hypertension (15 to 70%), ischemic heart disease (35%), psychological and psychiatric manifestations, pigmentation, and possible gonadal- and pregnancy-related effects. The primary lesion in analgesic nephropathy is renal papillary necrosis (RPN), and this is a nephrotoxic effect common to all nonsteroid antiinflammatory agents. The most important factor in the management of patients with analgesic nephropathy is the cessation of analgesic abuse, and this leads to improvement and stabilization of renal function. A small proportion of patients will, however, deteriorate in relation to accelerated hypertension, persistent proteinuria, ischemic heart disease, and complications leading to nephrectomy. Patients with analgesic nephropathy are poor risk patients and have a poor prognosis, even after dialysis and transplantation.


Shape of cells and intercellular channels in rabbit thick ascending limb of Henle. Electron micrographs of cortical thick ascending limb of Henle (TALH) were studied using morphometric techniques. The apical cell surface and the tubule basement membrane have identical areas of 0.8 x 10(5) mu2/mm of tubule length in a typical tubule (I.D. = 15 mu, O.D. = 25 mu). The total area of lateral cell walls bordering intercellular channels in 7.9 x 10(5) mu2/mm of typical tubule, and the ratio of apical cell surface to lateral surface in 0.10 +/- 0.01. When the photographed tubule mass was divided into five concentric zones of equal thickness, the lateral wall areas per zone were found to increase more rapidly than exponential, from 0.63 x 10(5) mu2/mm in that zone nearest the lumen to 3.6 c 10(5) mu2/mm in that zone adjacent to basement membrane. From these data and the estimated number of cells per mm of tubule length (764 cells), the circumferences of individual cells could be calculated for each zone, and quantative three-dimensional cell model could be constructed. The shape of intercellular channels is similar to that of the space between concentric, truncated, and plated horns. TALH cells are compared to previously described cells of rabbit proximal convoluted and straight tubules.


Histology of human tubulo-interstitial nephritis associated with antibodies to renal basement membranes. Twenty-seven patients with diffuse "crescentic" glomerulonephritis (CSGN) were identified in 1,174 renal biopsies from nephritic patients. Patients were assigned to three groups on the basis of the immunofluorescent study of renal biopsy specimens and serologic findings. Group I included eight patients with antibodies to glomerular (anti-GBM) and tubular (anti-TBM) basement membranes; group II had eight patients with only anti-GBM antibodies; and group III had eleven patients with CSGN unassociated with antibodies to either GBM or TBM. Patients with anti-GBM/anti-TBM antibodies (group I) had severe tubulointerstitial (TI) nephritis, as characterized by the infiltration of polymorphonuclear leukocytes and macrophages along the TBM and peritubular vessels. In some patients, focal proliferation of epithelial cells of proximal convoluted tubules (PCT), gaps or extensive destruction of TBM, lesions in the walls of small peritubular vessels, and interstitial giant cells were also observed. Patients with anti-GBM antibodies (group II) had mild to moderate interstitial cellular infiltration and mild tubular changes. Five patients with CSGN not associated with antibodies to renal basement membranes (group III) had mild to moderate interstitial cellular infiltration and tubular changes. A sixth patient, with Wegener's disease had severe granulomatous TI lesions. The results of this study show that TI nephritis is most frequent and severe with anti-TBM antibodies are demonstrable and suggest that anti-TBM antibodies contribute to the development of TI lesions.


Goodpasture's syndrome: an analysis of 29 cases. The pathologic features of 29 cases of Goodpasture's syndrome occurring during a 13-yr period in Auckland have been reviewed and correlated with clinical findings. There were 20 males and nine females in the series; two of the males and three of the females were Maoris. Age at the time of onset of symptoms ranged from 17 to 75 yr, with about 76% of the patients being from 17 to 27 yr of age. Sixteen (55%) of the patients died from less than a week up to about two years following the onset of symptoms, and the remaining 13 are live from 30 weeks to 14 yr after initial presentation. Underlying renal disease varied from mild focal glomerulitis to end-stage glomerulonephritis by light microscopy, but characteristic glomerular changes were seen in all specimens examined by electron and immunofluorescent microscopy. The lungs of 13 of the patients examined at autopsy showed typical abnormalities. The syndrome pursues a notably variable clinical course, affects a considerable proportion of females, occurs over a wide age range, and appears to be disporportionately common among Maoris.


Radioimmunoassay for urinary renal tubular antigen: a potential marker of tubular injury. A human proximal renal tubular epithelial antigen (designated HRTE-1) was isolated and purified from a crude tubular preparation (Fx1A) by a process of salt fractionation, DEAE anion-exchange chromatography, and Sephadex G-200 gel filtration. Utilizing 125I-HRTE-1 and a rabbit antiserum specific for the proximal tubular brush border, as determined by immunofluorescent microscopy, a radioimmunoassay by competitive protein-binding was developed. HRTE-1 was demonstrated in serum and urine and in extracts of a variety of body organs. A range of concentrations for normal random urine samples and 24-hr urine excretion rates were determined. Random urine samples from 36 patients with a variety of functional and pathologic renal disorders were assayed for the HRTE-1 antigen. Twenty-three of 24 patients with either chronic nephropathy or pre-renal azotemia had normal urinary antigen concentrations, despite wide differences in urine flow rates, the degree of existing renal function, and the amount of proteinuria. Ten of 12 patients with acute tubular necrosis, however, had statistically abnormal HRTE-1 concentrations (high in eight patients, undetectable in two). These findings suggest that HRTE-1 antigen can be detected in both normal and pathologic urines, that altered antigen concentrations can be documented in at least one renal disorder, and that quantitation of HRTE-1 in urine may have clinical value as a marker of acute rubular injury.


[Duchenne-type muscular dystrophy: problems, early diagnosis, early treatment (author's transl)]. The aetiology and pathogenesis of the Duchenne-type muscular dystrophy (pseudohypertrophic muscular dystrophy) are still largely unknown. The possibilities of treating the disease are rather limited. Treatment is the more successful, the earlier diagnosis was possible, and the earlier treatment is initiated. The CK-Screening Test is an important aid for early diagnosis. The CK-Screening Test is also valuable for genetic consultation and advice, because it helps to identify women who are conductors or carriers of the disease. Current hypotheses on aetiology and pathogenesis are mentioned. Progress made in the fields of biochemistry, including enzyme histochemistry, and electron microscopy, raise hopes of finding more efficient therapeutic possibilities in the future. The many interests of patients with muscular diseases are being looked after by the European Alliance of Muscular Dystrophy Associations (EAMDA). Thirteen European associations are members of this organisation, including the German association "Bekämpfung der Muskelkrankheiten e.V." The number of sponsoring members of the EAMDA is at present about 300,000. 3 international congresses have already been held on the problems of muscular diseases. The fourth congress is scheduled to take place in Montreal in 1978.


[Clinical pharmacokinetics of diazepam and its biologically active metabolites (author's transl)]. The pharmacokinetics of diazepam and its biologically active metabolites desmethyldiazepam and oxazepam is critically evaluated from a clinically relevant point of view. The slow elimination of diazepam is dependent on the degree of plasma protein binding, the duration of the medication, the age and the liver function of the patient. While the normal half-life (T1/2(beta)) varies between 1 and 2 days, it can be increased to up to 80--100 h in subjects over 60 years of age. In patients with liver disease T1/2(beta) is about doubled, which is caused by a reduction (factor 2) of the normal hepatic clearance of 26 ml/min. After subchronic treatment with diazepam the elimination rate is reduced about 20--70% in healthy subjects, but liver patients exhibit only a slightly further prolongation in T1/2(beta). The major metabolite desmethyldiazepam has a T1/2(beta) of 51 h and a Cl of 11 ml/min and accumulates after multiple doses of diazepam since its elimination is much slower than that of its parent compound. The elimination of this drug is also impaired (factor 2) in patients with liver disease. In contrast to these findings oxazepam is excreted as glucuronide in the urine relatively fast and independently of the liver function with a T1/2(beta) of 5.5 h and a Cl of 130 ml/min.


[The agar culture of the leucopoietic stem cell (CFU-C) and its stimulation (author's transl)]. This review summarizes the different in vitro culture techniques of the leukopoietic stem cell. Mainly the results obtained by the system of Pluznik and Sachs and Bradley and Metcalf are described apply with semisolid agar and its modification by Iscove using methylcellulose. Furthermore, recently published modifications of culture technique are listed. The principal results obtained with mouse and human bone marrow cells are summarized with respect to the origin of the colony-stimulating factor acting in different species, its biochemical characteristics, as well as with regard to the colony forming cell and its different subclasses.


[Phagocytosis--a central mechanism in inflammatory reaction (author's transl)]. It is attempted to describe the complex pathogenesis of the inflammatory reaction by analysing the mechanisms of several reactions involved. Phagocytosis is a central phenomenon of host defense mechanisms responding to an invasion of foreign bodies. The induction of secondary reactions following phagocytosis is looked at. Several pathogenetic mechanisms responsible for a switch from acute to chronic inflammation are suggested. Known principles of the autonomous regulation of cell effector functions are discussed in relation to their participation in inflammatory reactions.


On the terms "reticulosis" and "reticulum cell sarcoma" with regard to the modern concept of the monocyte macrophage system. The terms "reticulosis" and "reticulum cell sarcoma" (=malignant lymphoma, histiocytic type) are discussed regarding the modern concept of the monocyte macrophage system which today has replaced the ancient theory of the reticuloendothelial system. The monocyte macrophage system which today has replaced the ancient theory of the reticuloendothelial system. The monocyte macrophage system is not independent, but closely related to the myeloid system. Thus, a third blood forming system as was believed in the case of RES does not exist. Phagocytic reticulum cells of the various henatopoietic organs are highly activated monocyte-derived macrophages. All those conditions formerly termed "reticulosses" have been found to belong either to the myeloid or to the lymphatic system. Considering the reticulum cell sarcomas or malignant histiocytic lymphomas, most of them seem to be of lymphatic rather than of macrophage origin, representing highgrade malignant lymphomas, possibly immunoblastic sarcomas. No relationship between these tumours and the monocyte macrophage system has been established, so far. Therefore, the terms "reticulosis" and "reticulum cell sarcoma" should be no longer used in order to avoid confusion, in order to stimulate sufficient diagnostic efforts which will really clarify such cases, and in order to give full credit to modern results of hematopathology.


Thyroid disease in relation to breast cancer. A controversy exists in regard to thyroid function and breast cancer. Hypothyroidism has been suggested as being either protective from breast cancer or predisposing to the disease. It has been hypothesized that a deficiency in circulating thyroid hormones may hypersensitize the mammary glandular epithelium toward prolactin and estrogens, thus aiding the development of breast neoplasia. On the other hand, thyroid hormone replacement therapy has been connected with an increased risk of breast cancer, but this has been contested. At this time the American Thyroid Association recommends that, if indicated, hypothyroid patients should take their thyroid hormone medication. Hyperthyroidism has been associated with a decreased risk of breast cancer. Also, in hyperthyroid patients with inoperable breast cancer, the malignant growth is thought to be slowed. However, this, too has been disputed. Moreover, hyperthyroidism has been connected with the development of breast cancer in premenopausal women. At present no role of thyroid hormone in the pathobiology of breast cancer can be defined. It seems that the "thyroid-breast cancer controversy" can only be resolved by a prospective study preferably on postmenopausal women correlating thyroid (T3, T4, PBI), pituitary (TSH, TRH, Prolactin), and adrenocortical (androgens) function tests with the clinical examination of thyroid, breast, and genital apparatus and determination of the estrogen status (vaginal smear, plasma estrogens) as well.


Morphology of the early hemostasis in human skin wounds: influence of acetylsalicylic acid. The in vivo formation of hemostatic plugs was studied in humans in skin wounds made using the template bleeding procedure of Mielke (34). The wounds were excised by punch biopsy 10 seconds, 30 seconds, 2 minutes, and 3 minutes after they were made. The wounds were V-shaped and approximately 0.4 mm. deep. Within 30 seconds small hemostatic plugs were observed at the end of transected vessels. The plugs grew in size in the subsequent minutes and became impermeable. The platelets degranulated and formed pseudopods which became strongly interdigitated. The platelets at the periphery of the plugs showed discontinuities of the membranes. Cytoplasmic matrix and cell organelles had disappeared in many of these peripheral cells. Small fibrin fibers were already found at 30 seconds, mostly along the margins of the wounds and also at the periphery of the hemostatic plugs. Fibrin was absent from the center of the plugs and from the lumen of transected vessels. When part of a plug was extending into the vessel lumen, the platelets inside the vessel were less degranulated and less interdigitated than the rest of the plug. The effect of acetysalicylic acid (ASA) was studied in wounds before and 2.5 hours after ingestion of 2 gm. of ASA. Wounds were excised by punch biopsy 3 or 10 minutes after they had been made. Platelets in ASA were less degranulated, had fewer pseudopods, and showed less interdigitation than platelets in control plugs. Ballooning and fibrin deposition were similar in control and ASA plugs. Pronounced differences between control and ASA plugs were observed in a subject who exhibited a considerably prolonged bleeding time after ASA. The ASA plugs were very large; many plugs had fused and in addition numerous small platelet clumps, most likely fragments from the plugs, were found in the superficial scab of the wound. It is postulated that ASA plugs are less stable due to decreased interdigitation. This allows more disruption of the plugs and rebleeding. Consequently, more platelets are needed and longer time is required for hemostasis to occur.