Purification and some properties of tartrate-sensitive acid phosphatase from rabbit kidney cortex. Two forms of tartrate-sensitive acid phosphatases (EC 3.1.3.2) were purified from rabbit kidney cortex by a multiple-column-chromatography method. The basic form constituted 90% of the enzyme and migrated as a single band of protein on polyacrylamide-gel electrophoresis. The proteins contaminating the acidic form did not exceed 5% of the total protein. The specific activity towards p-nitrophenyl phosphate was 12 mumol/min per mg for the basic form and 0.7 mumol/min per mg for the acidic form. The basic form of the enzyme differs from the acidic form in its heat-stability, Km values, inhibition rates by tartrate and fluoride and substrate specificities. Relative to p-nitrophenyl phosphate hydrolysis rate, the acidic form hydrolysed a variety of physiological monophosphate esters, whereas the basic form hydrolysed only CMP and phosphoenolpyruvate. Bacterial neuraminidases had no effect on the activity and mobility of the acidic form on polyacrylamide-gel electrophoresis. Both forms have the same molecular weight (101000 +/- 4000) and are probably composed of two identical subunits. The question whether the two forms of the enzyme are different proteins or whether one is a modified form of the other is discussed.


Dissociation of fibronectin from gelatin-agarose by amino compounds. Soluble fibronectin of human plasma was specifically dissociated at neutral pH from gelatin-agarose by several cationic amino compounds, notably the polyamines spermine, spermidine and putrescine, the basic amino acid arginine, and amino sugars. The neutral and acidic amino acids and the N-acetylated derivatives of amino sugars tested were ineffective. Gel-filtration experiments demonstrated that [14C]spermidine bound to fibronectin but not to gelatin.


Strong metal-oxygen interaction in uracils. X-ray crystal structure of bis-(1,3-dimethyluracil)dichlorocopper(II). An X-ray structure determination showed that in the compound bis-(1,3-dimethyluracil)-dichlorocopper(II) the uracil moiety is strongly bonded to the metal via the O(4) atom. Changes in the i.r. spectra on metal binding are discussed. Details of the structural parameters have been deposited as Supplementary Publications SUP 50091 (3 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms indicated in Biochem. J. (1978) 169, 5.


Intermolecular specificity of pancreatic lipase in the lipolysis of triacylglycerols containing phytanic acid. Treatment of mixtures of glycerol tripalmitate and rac-glycerol 1,2-dipalmitate 3-phytanate with pancreatic lipase demonstrated that phytanic acid retarded the release of palmitic acid esterified in the same triacylglycerol.


Incorporation of [18O]water in the formation of p-hydroxybenzyl alcohol by the p-cresol methylhydroxylase from Pseudomonas putida. In the hydroxylation of the methyl group of p-cresol by an enzyme from Pseudomonas putida the oxygen atom is derived from water. Although a second reaction by the same enzyme converts the product, p-hydroxybenzyl alcohol, into the aldehyde, the alcohol is an enzyme-free intermediate.


The isolation and partial characterization of low-molecular-weight phosphorylated component of the non-histone proteins of mouse nuclei. After labelling of mouse liver nuclei with [gamma-32P]ATP in vitro, 10-20% of the radioactivity incorporated into the saline-soluble nuclear and HAP2 chromatin fractions was located in a low-molecular-weight component (component 10) with pI near 4.5 in urea. By using combinations of ion-exchange chromatography, preparative thin-layer isoelectric focusing and gel filtration, this component was isolated from both nuclear fractions. Recovery from the saline-soluble fraction was poor under conditions that allow endogenous phosphatases to be active. Component 10 was shown to be a phosphoprotein on the basis of enzyme-digestion experiments and the detection of phosphoserine and phosphothreonine. The 32P radioactivity did not appear to be associated with phosphorylated basic amino acids. Its molecular weight was determined by gel chromatography and electrophoresis in sodium dodecyl sulphate/polyacrylamide gels as approx. 10000, and tryptic digestion of the reduced carboxymethylated protein in urea yielded two 32P-labelled peptides. It has not been possible as yet to assign a function to component 10, though its similarity to other low-molecular-weight acidic proteins is discussed.


Polypeptide linkages and resulting structural features as powerful chromogenic factors in the Lowry phenol reaction. Studies on a glycoprotein containing no Lowry phenol-reactive amino acids and on its desialylated and deglycosylated products. With bovine serum albumin as the reference standard, the armadillo salivary-gland glycoprotein, although containing no chromogenic amino acids and only small amounts of colour-yielding peptides [Chou & Goldstein (1960) Biochem. J. 75, 100-115], is highly reactive in the Lowry phenol protein assay [Wu & Pigman (1977) Biochem. J. 161, 37-47]. After desialylation and Smith degradation of the glycoprotein, the Lowry phenol value increased by 13 and 30% respectively, which suggests that both sialic acid and N-acetylhexosamine exert shielding effects in this reaction. Acid hydrolysis for 30 min decreased the Lowry phenol value by more than 45%, which indicates that the peptide linkages and steric features affect the Lowry phenol reactivity. After hydrolysis for up to 6h, the remaining Lowry phenol value of the partially hydrolysed core protein paralleled the amount of unhydrolysed peptides, inferring that both acid-sensitive and acid-resistant chromophoric peptides are fairly evenly distributed along the whole polypeptide chain. As with bovine serum albumin, more than 80% of the colour yield obtained in the Lowry phenol assay with this glycoprotein is Cu2+-dependent.


Studies on the interaction between disulfiram and sheep liver cytoplasmic aldehyde dehydrogenase. The effect of disulfiram, [1-14C]disulfiram and some other thiol reagents on the activity of cytoplasmic aldehyde dehydrogenase from sheep liver was studied. The results are consistent with a rapid covalent interaction between disulfiram and the enzyme, and inconsistent with the notion that disulfiram is a reversible competitive inhibitor of cytoplasmic aldehyde dehydrogenase. There is a non-linear relationship between loss of about 90% of the enzyme activity and amount of disulfiram added; possible reasons for this are discussed. The remaining approx. 10% of activity is relatively insensitive to disulfiram. It is found that modification of only a small number of groups (one to two) per tetrameric enzyme molecule is responsible for the observed loss of activity. The dehydrogenase activity of the enzyme is affected more severely by disulfiram than is the esterase activity. Negatively charged thiol reagents have little or no effect on cytoplasmic aldehyde dehydrogenase. 2,2'-Dithiodipyridine is an activator of the enzyme.


Heterogeneity in horse ferritins. A comparative study of surface charge, iron content and kinetics of iron uptake. Horse ferritins from different organs show heterogeneity on electrofocusing in Ampholine gradients. Both ferritin and apoferritin from liver and spleen could be fractionated with respect to surface charge by serial precipitation with (NH4)2SO4. In the ferritin fractions, increasing iron content parallels increasing isoelectric point. After removal of their iron, those fractions which originally contained most iron accumulated added iron at the fastest rates. When unfractionated ferritins from different organs were compared the average isoelectric point increased in order spleen less than liver less than kidney less than heart. The order of initial rates of iron uptake by the apoferritins was spleen greater than kidney greater than heart and initial average iron contents also followed this order. The relatively low rates of iron accumulation by iron-poor molecules may have been due to structural alteration, to degradation, to activation of the iron-rich molecules or to other factors.


Studies on immune complexes in rheumatoid arthritis. Sera and synovial fluids of patients with rheumatoid arthritis were studied for immune complexes by means of the anti-antibody inhibition test; approximately 40% of these specimens were shown to contain immune complexes. To study the nature of antigens participating in the formation of complexes under investigation, the anti-antibody inhibition test was repeated after addition of various soluble antigens to the complex-positive rheumatoid arthritis sera. This resulted in conversion of some sera from inhibitory to noninhibitory. It was found that of 15 positive rheumatoid arthritis sera, 7 had complexes formed by heterophile, Hanganutziu-Deicher antigen and of 12 positive sera tested, 2 had complexes formed by human IgG.


A study of HLA antigens in adults with acute rheumatic fever. By use of a microlymphocytotoxic assay to detect 32 HLA antigens of the A and B series, we examined 49 Mexican-American adults with acute rheumatic fever and contrasted the findings with 100 ethnically identical controls. Eighty-nine percent of the total possible antigens were identified. Strong associations between HLA antigens from either locus and acute rheumatic fever were not detected. These findings serve to emphasize the importance of nongenetic factors in the pathogenesis of acute rheumatic fever and help clarify the role of HLA antigens in rheumatic diseases.


Androgenic status of males with systemic lupus erythematosus. The female predominance among patients with systemic lupus erythematosus and the protective effects of androgens in the murine lupus model prompted us to assess the androgenic status of 12 men with the disease. Save for minor abnormalities explained by treatment or renal insufficiency, normal serum testosterone, luteinizing hormone, and follicle stimulating hormone levels were found. Hypogonadism or androgen deficiency was not evident.


Antigen-induced arthritis. Decreased proteoglycan content and inhibition of proteoglycan synthesis in articular cartilage. An arthritis was induced in rabbits immunized with human serum albumin by injection of antigen into the hind knee joint. Histological changes in the synovial membrane and an increase in polymorphonuclear granulocytes in the synovial fluid indicated an inflammatory response similar to that described with fibrin as antigen (1). The arthritis was accompanied by no significant change in the collagen content, but a marked decrease in the proteoglycan content of the cartilage was noted. Cartilage from inflamed joints generally exhibited a decreased ability to synthesize proteoglycan in vitro.


Ocular chrysiasis correlated with gold concentrations in the crystalline lens during chrysotherapy. The eyes of 11 patients with rheumatoid arthritis (RA) who received extended chrysotherapy (mean cumulative dose greater than 7 grams during a mean 6-year period) were examined biomicroscopically. Minute reddish-purple particles were seen in the cornea (corneal chrysiasis) in 5 and in the lens (lens chrysiasis) in 4 patients. Particulate deposits were absent in 11 other RA patients who had not received gold treatment. Seven crystalline lenses from 5 gold-treated patients were removed surgically because of incidental cataract formation and analyzed for gold content using neutron activation analysis. Although the mean lens gold concentration was higher in these patients than in non-gold-treated controls without RA (0.0073 microgram/grams versus 0.001 microgram/grams), the absolute gold level was markedly lower than that found in 25 diverse tissue analyzed previously. This finding is compatible with the absence of clinical gold-related lens disease or visual impairment.


Interrelationship between stimulation of prostaglandin E and hyaluronate production by poly (I) . poly (C) and interferon in synovial fibroblast culture. The interferon-inducer, polyinosinate . polycyctidylate [Poly (I) . Poly (C)] stimulated both prostaglandin E (PGE) and hyaluronic acid (HA) production by human cultured synovial fibroblasts. Increased accumulation of PGE in the medium was noted within 3 hours of culture, while accumulation of HA was observed only after a 24 hour lag period. The stimulation of PGE and HA production by Poly (I) . Poly (C) was prevented by aspirin and indomethacin. Human interferon was also effective in stimulating both PGE and HA production. Concomitant addition of cortisol to culture medium abrogated the stimulatory effect of Poly (I) . Poly (C) and interferon on both PGE and HA production. Exogenous PGE2, however, overcame the inhibitory effect of cortisol on HA production. The present results suggest that the stimulatory effect of both interferon and Poly (I) . Poly (C) on HA production may be mediated by PGE. Prostaglandins may thus play a role in the inflammatory process associated with viral infections.


The amyloidosis of juvenile rheumatoid arthritis--comparative studies in Polish and American children. I. Levels of serum SAA protein. Serum SAA concentration was determined by radioimmunoassay in 21 Polish children with amyloidosis secondary to juvenile rheumatoid arthritis (JRA). The results were compared to controls and children with JRA in Polish populations (where amyloidosis is a frequent complication of JRA) as well as to American children with JRA (where amyloidosis in JRA has been observed only sporadically) and American control children. No significant differences of SAA protein levels were found in the amyloidotic Polish children when compared to JRA Polish and American children. However, significantly higher levels of SAA protein were present in amyloidotic Polish children when compared to the control Polish and American group. High serum SAA protein concentration in JRA children did not necessarily correlate with the presence of secondary amyloidosis. Other mechanisms are probably involved in the development of amyloidosis.


The satellite arthritis clinic.

The satellite arthritis clinic. For nine years an Arthritis Consultation Clinic has served a large community that did not have a rheumatologist. A university-based rheumatology section from another city, a local hospital, the United Way, a chapter of the Arthritis Foundation, and the County Medical Society have all contributed to the operation of the clinic. Such a clinic may be viewed as a stage in the regional development of rheumatologic care. Although primarily intended as a community service program, the clinic may also be useful in professional education on several levels.


Marrow aplasia induced by colchicine. A case report. A 70 year old male developed marrow aplasia and pancytopenia after receiving 10 mg of colchicine intravenously over a period of 5 days for acute gouty arthritis. Despite a fatal outcome, evidence of recovery of hematopoiesis was observed in the bone marrow 3 days after the last dose of the drug. The pathogenetic mechanism by which colchicine induced marrow aplasia in this patient remains unclear.


HLA-C locus antigens in HLA-B27 associated arthritis. Frequencies of the HLA-C locus antigens, Cw1, Cw2, Cw3, and Cw4, were determined in 88 patients with spondylitic diseases and 88 matched and 64 B27-positive normal controls. Both Cw1 and Cw2 were significantly increased in B27-positive patients and B27-positive controls as compared to B27-negative patients and matched controls. Cw1 and Cw2 demonstrate strong linkage disequilibrium with B27. Their absence in B27-negative patients suggests disease susceptibility is related to HLA-B.


Involvement of prostaglandins from rheumatoid synovium in inhibition of articular cartilage metabolism. Short term incubations of articular cartilage in media from cultures of rheumatoid synovial tissue result in marked inhibition of 3H-glycine and Na235SO4 incorporation into trichloroacetic acid (TCA) insoluble macromolecules by the cartilage segments with little effect on degradation of prelabeled cartilage matrix. This inhibition is time dependent and not observed in cartilage segments incubated in media derived from rheumatoid synovia cultured in the presence of indomethacin (a specific inhibitor of prostaglandin synthesis). In addition, no indomethacin-sensitive cartilage inhibitory activity was detectable in media from cultures of normal synovia. The ether solubility of the majority of the inhibitory activity and the indomethacin sensitivity suggest that the inhibitor(s) are prostaglandins.


High prevalence of HLA-Cw1 and Cw2 antigens in spondylarthritis. HLA-B27 is frequently, but not invariably, found in patients with spondylarthritic diseases. Recently it has been shown that HLA-C-locus antigens w1 and w2 (w1/2) are in linkage disequilibrium with B27. A group of 139 patients with seronegative arthritis or spondylitis has been studied to determine the clinical and radiographic features associated with these antigens. An increased frequency of spondylarthritis was present in HLA-B27-negative, Cw1/2-positive patients when compared with B27- and Cw1/2-negative patients (P less than 0.01). In addition, a large group of patients who did not fulfill accepted criteria for a specific spondylarthritic syndrome were identified; the frequency of both HLA-B27 and Cw1/2 was greater in these individuals than in healthy blood donors. Thus HLA-Cw1/2 are spondylitis-associated antigens. These findings suggest that the critical genetic locus is neither B27 nor Cw1/2 but is a closely linked gene.


Complement-fixing hidden rheumatoid factor in juvenile rheumatoid arthritis. Fifteen to twenty percent of patients with juvenile rheumatoid arthritis (JRA) have positive latex fixation tests (LFT), whereas approximately 46% have previously been demonstrated to have hidden rheumatoid factors (RF), i.e., 19S IgM RF which can be detected by the LFT after acid separation of the IgM-containing fraction from serum. In this study, hidden RF were found in 59% of patients with seronegative JRA by use of a complement-dependent hemolytic assay. The median titer of JRA patients was 1:42, and in healthy and disease controls it was 1:7. The difference was significant at P less than 0.001. When data from patients with active disease were analyzed separately, the median titer for polyarticular JRA was 1:97 and for pauciarticular JRA, 1:91. The differences due to active disease were significant at P less than 0.001 and P less than 0.005, respectively. The results demonstrate that the hemolytic assay is more sensitive than the LFT in determining the presence of hidden RF, and activity of disease correlates well with high hemolytic RF titers.


Fate of antigen after intravenous and intraarticular injection into rabbits. To determine the manner in which small protein antigens enter the synovium and are localized, horseradish peroxidase (molecular weight 40,000) was injected by intravenous and intraarticular routes. Passage of antigen from capillary lumen to joint space and vice versa occurred primarily across fenestrated vessels. Antigen localized to collagen fibers and lining cells. At low concentrations of antigen, antigen was taken up predominantly by marcophages, whereas at higher concentrations both macrophages and fibroblast-like lining cells accumulated antigen.


Mechanisms of cellular interaction with monosodium urate crystals. IgG-dependent and IgG-independent platelet stimulation by urate crystals. Monosodium urate crystals (MSU) stimulate suspensions of washed platelets or neutrophils. When MSU crystals are coated with IgG, as occurs in plasma, stimulation is markedly enhanced. These studies which use MSU-induced human platelet serotonin secretion as a model examine the nature of cellular recognition mechanisms for MSU crystals and IgG-coated MSU crystals. F(ab')2 fragments of specific anti-Fc antibody blocked and the lipopolysaccharide of Salmonella minnesota R595 enhanced human platelet secretion induced by IgG-coated urate crystals. These agents had little effect on stimulation by uncoated crystals. This indicated that urate crystals stimulate platelets independently of fluid phase IgG. Urate crystals directly stimulated suspensions of washed rabbit platelets which lack Fc receptors. In contrast to human cells, stimulation was blocked by IgG. This again demonstrated IgG-independent cell stimulation by urate crystals. Calcium pyrophosphate dihyrate crystals could trigger human platelet secretion only when coated with IgG. This suggests that when crystals are coated with IgG, the surface-bound IgG alone may be the stimulus to the cell. This was supported by the finding that polyvinylpyridine-N-oxide, a hydrogen acceptor, blocked human platelet stimulation by uncoated, but not IgG-coated, urate crystals. These data indicate that urate crystals (and potentially other surface or particles) can stimulate a mediator cell by at least two mechanisms: by direct stimulation without the mediation of adsorbed IgG or, when coated with IgG, by triggering the cell via immunoglobulin receptors.


Effects of diphosphonates on glycosaminoglycan synthesis and proteoglycan aggregation in normal adult articular cartilage. The effects of disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP) and disodium dichloromethylene diphosphonate (Cl2MDP) on glycosaminoglycan synthesis and macromolecular organization of proteoglycans have been examined in normal adult canine cartilage. One to 500 micron of either compound produced reversible inhibition of glycosaminoglycan synthesis to about 70% of control levels, whereas lower concentrations had no effect. Based on Sephadex G-200 chromatography, the average hydrodynamic size of glycosaminoglycans in diphosphonate-treated cartilage was similar to that of those in control cartilage. In most cases proteoglycan aggregates from diphosphonate-treated cartilage were smaller in hydrodynamic size than those from control cartilage, as judged by Sepharose 2B elution profiles. The size of purified proteoglycan subunits, obtained after dissociation of the aggregates with 4 M guanidinium chloride or after incubation of the aggregates with hyaluronic acid beta1 leads to 3 hydrolase, was not affected by the diphosphonates. Furthermore, proteoglycans from diphosphonate-treated cartilage did not interact in vitro with hyaluronic acid, suggesting that diminished proteoglycan aggregation may have resulted from an abnormality in the hyaluronate-binding region of the proteoglycan core protein.


Familial deficiency of two subunits of the first component of complement. C1r and C1s associated with a lupus erythematosus-like disease. Complete absence of C1r and almost complete absence of C1s were found in 4 of 8 living siblings. Two of the 4 suffer from a syndrome that combines discoid lupus erythematosus and nondeforming rheumatoid-like arthritis; one of the siblings has mild nephritis. The other 2 C1 deficient family members are clinically well. Evidence from this and other families suggests that deficiency of C1 components or C4 is associated with higher risk of developing a lupus-like disease than is deficiency of C2.


Complement-fixing hidden rheumatoid factor in children with benign rheumatoid nodules. Rheumatoid nodules have been described in many children without evidence of clinical disease. These have been referred to as benign rheumatoid nodules (BRN). Except for one report, no serological abnormalities have been demonstrated. Four children with BRN have been studied. By means of a hemolytic assay, high titers of hidden rheumatoid factor (RF) have been found in the blood, i.e., 19S IgM RF detected after acid separation of IgM-containing fraction from the serum. The median hidden RF titer of the children with BRN was 1:362 and in healthy and disease controls was 1:7. The difference was significant at P less than 0.001. The results indicate that patients with BRN have active complement-fixing 19S IgM RF in their serum. Thus the association of hidden RF with BRN raises the possibility that the deposition of immune complexes (19S IgM-7S IgG) leads to small vessel vasculitis and nodular formation.


Acute hematogenous osteomyelitis progressing to septic synovitis and eventual pyarthrosis. The vascular pathway. A case suggesting sequential infection of bone, synovial membranes, and finally synovial fluid is presented. Hematogenous osteomyelitis manifests itself differently in children than in adults. In children, the avascular epiphyseal cartilage in the growing long bone is an effective barrier to the spread of bone infection to the joint. After obliteration of the growth plate, vascular anastomoses form between metaphysis and epiphysis, and at maturity the entire bone shares a common blood supply with the capsule and synovium of the adjacent joint. In the adult with acute osteomyelitis, contiguous joint infection should be anticipated. In septic arthritis, unless caused by penetrating trauma, the synovium must be infected before the joint fluid. Even when bacterial joint infection is present, repeated arthrocenteses may be required over several days to confirm the diagnosis.


Legionnaires' disease in a patient with systemic lupus erythematosus. A young woman with systemic lupus erythematosus developed a rapidly fatal pneumonia from which no visible or culturable organisms were found. Subsequent stains disclosed typical findings of Legionnaires' disease. A cutaneous portal of entry was suspected and a fulminant lung abscess developed, neither of which has been previously reported in Legionnaires' disease.


Echovirus-associated polyarthritis. Report of a case with synovial fluid and synovial histologic characterization. A patient with fever, myalgias, and acute polyarthritis in whom echovirus 9 was isolated from throat and rectal swab specimens is described. All symptoms resolved spontaneously over a one week period except for the arthritis which required three months for complete recovery. A mildly inflammatory synovial fluid and histologic picture were observed. Attempts to isolate the virus from synovial fluid and synovium during acute illness were unsuccessful.


Transverse myelitis in mixed connective tissue disease. Neurologic disease is reported to occur in just 10% of patients with mixed connective tissue disease (MCTD). Most commonly, this is manifested by mild trigeminal neuralgia. This report details the clinical and neuropathologic findings of transverse myelitis in a patient with MCTD. Neurologic features include progressive areflexic paraplegia with loss of bowel and bladder function. Neuropathologically there was thinning of the thoracic cord, widespread loss of axons and myelin sheaths, reactive astrocytosis, macrophage formation, vascular thickening with perivascular chronic inflammatory cell infiltration, and calcium deposits. This case demonstrates that severe neurologic disease unresponsive to therapy can occur in MCTD.


[Use of an immunomodulator (levamisole) in HbsAg positive patients]. The results of some clinical trials performed with levamisole on 12 HBsAg-positive subjects, including 5 patients with aggressive chronic hepatitis, (ACH) 2 patients with persistent chronic hepatitis (PCH) and 5 healthy carriers are reported. Levamisole was administered in 2.5 mg/Kg/day doses for three consecutive days. During treatment prothrombin activity normalized in ACH and PCH as well as transaminases, the latter starting from the 4th week, even though a two-fourfold increase of the starting values was observed in the first weeks of disease. HBsAg, anti-HBs titres and immunocomplexes values did not show any significant variations, but for 1 case, while some immunological parameters (E, EA rosettes) normalized in those cases showing base- values lower than the norm.


Correlations between anti-dsDNA antibodies, complement levels and clinical course in SLE patients treated with steroid pulse therapy. In patients with SLE dsDNA binding activity, C3 and C4 levels are correlated with the clinical course of the nephritis. In fact the study of these serological parameters has a prognostic value in patients treated with methylprednisolone pulse therapy. This treatment can rapidly normalize an acute phase.


[Hepatitis B and non-B in our case reports of post-transfusional hepatitis]. The Authors examined 52 patients with post-transfusional hepatitis admitted in the Insitute of Infectious Diseases in Perugia, from 1971 to 1977. 30 (57.7%) patients had "B" hepatitis, 22 (42.3%) "non B" hepatitis. The incidence of anti-HBs antibodies in the two groups of patients suggests that they are partially protective against "B" hepatitis. "B" and "non B" hepatitis resulted clinically different only in the incubation period, 95.6 days (mean) for "B" hepatitis and 62.1 days (mean) for the "non B" hepatitis.


[Salmonellosis in Cagliari].

[Salmonellosis in Cagliari]. The therapy (generally ineffective) and the prophylactic measures of "Salmonellosis" are examined critically during a period of three years, 1974 -- 1976, when a large increase of infection was present in children (without letal cases). The extension of mass-vaccination, particularly by oral treatment with microbic pap of attenuate living (or dead) stock that represent the most significant antigenic groups (4, 9, 3, 6, 7, 8) is suggested.


[Epidemiological surveillance of influenza in western Sicily in 1976-1977]. Two strains of influenza virus A/H3N2 antigenically similar to A/Victoria/3/75 have been isolated in Palermo in February 1977; a peak of extramortality has been observed in the same month, and a rise of HAI antibodies for A/H3N2 viruses in human serum samples, taken from apparently healthy subjects living Palermo, has been demonstrated between October 1976 and February 1977. Similar rise of antibody titers has also taken place in serum samples from two communities in Western Sicily (Favara, 30,000 inhabitants, Favignana, 3,000 inhabitants). Serum samples obtained in June 1977 showed a slight fall of antibody level in Palermo and in Favignana, whereas in Favara a further rise, in comparison to February, is consistent with the hypothesis of a delayed circulation of virus in that community. Reduction of HAI titers for H3N2 viruses was even more pronounced in October 1977 in all groups of serum samples tested. The importance of local surveillance of influenza and the assessment of herd immunity level is discussed.


[Electrophysiological responses of cerebellum after the administration of diphenylhydantoin (author's transl)]. Experiments were performed to see the mechanism of anticonvulsive action of Diphenylhydantoin (DPH) especially in relation to the cerebellum. 15 normal adult cats were divided into three groups. After the intravenous administration of 10 mg, 20 mg and 30 mg/kg of DPH in each group, electrophysiological responses of the cerebellum with regard to Purkinje cell action potentials and the cerebellar electrocorticogram were observed for two hours. Purkinje cell action potentials were evaluated in its discharge rates, which were calculated with medical computer. The cerebellar electrocorticogram were analyzed with Fast Fourier Transform method especially in its Adrian rhythm (150--250). It has been demonstrated that Purkinje cell discharge rates increased extremely after a transient suppressed phase, but fast waves of the cerebellar electrocorticogram showed no remarkable changes following the administration of DPH. Plasma DPH concentration went up to  a toxic level immediately after DPH injection, but returned to the plateau level after ten to thirty minutes which gradually decreased for two hours. Distribution of DPH was always higher in the cerebellum than in the cerebrum. Recent experimental and clinical studies are suggesting that the cerebellum acts as one of the important intrinsic inhibitory system against epilepsy. Our results seem to imply that DPH exerts its anticonvulsive effect by potentiating inhibitory mechanism of Purkinje cells over the cerebral cortex. The cerebellar electrocorticogram and the Purkinje cell unit activities showed no definite correlative responses to DPH administration. This will suggest that cerebellar electrocorticogram is not the only reflection of the Purkinje cell activity but the total electrophysiological phenomena of the cerebellar cortex.


Analysis of regional left ventricular wall movement by phased array echocardiography. Images from standard two-dimensional echocardiographs do not lend themselves to the study of regional left ventricular wall movement because of poor definition of endocardium. An alternative method, based on a wide-angle phased array sector scanner has, therefore, been devised. Once the appropriate region of the heart has been identified, the scan rate is increased, and an array of 41 M-mode scans is produced, resulting in improved frequency response and endocardial definition. Each scan is digitised separately, and since their spatial orientation is known, the left ventricular image can be reconstituted by computer, and isometric or contour displays produced. Preliminary results, based on 40 patients, are presented showing examples of left ventricular wall movement. Regional abnormalities of amplitude or timing can be displayed with results similar to those of angiography, while disturbances of septal motion or wall thickness can be observed in greater detail than with other methods.


A method for praecordial surface mapping of the exercise electrocardiogram. A technique for praecordial surface mapping of the exercise electrocardiogram is described. This showed the area, time course, and severity of ST segment depression as projected onto the front of the chest after exercise. Twenty normal volunteers and 20 patients with coronary artery disease have been studied. No changes were seen after exercise in the normal subjects but areas of ST segment depression appeared in all 20 patients with angina pectoris. In 5 of the 20 patients with coronary artery disease, the exercise test was repeated on a later date. There were no significant differences in the area of severity of electrocardiographic abnormalities recorded during the two tests. This technique may prove to be useful for diagnosis and assessing medical and surgical treatments in patients with ischaemic heart disease.


Effects of recording speed on precision of time-based polycardiographic measurements. Optimal paper speeds for measuring points and intervals. Optimal paper speeds have not been established for all time-based measurements of the cardiac cycle by appropriately designed observer performance studies. In 10 subjects (5 normals and 5 cardiac patients) carotid pulse, phonocardiogram, and electrocardiogram were recorded on magnetic tape for measurement of all fiducial points for systolic time intervals, the systolic time intervals themselves, the pulse transmission time, cycle length (RR), qR time, and R-to-point versus q-to-point measurements at recording speeds of 25, 50, 75, 100, and 200 mm/s. Tracings were coded numerically and randomised. Three observers measured all points and calculated intervals in a sequence determined by individual tables of random numbers. Left ventricular ejection time was the only calculation that could be made at 25 mm/s statistically equally well as at all other speeds. The smallest numerical observer differences occurred uniformly at 100 mm/s paper speed when all recording speeds were considered. However, after excluding the 25 mm/s speed there were no significant differences among point measurements. Measurements of points from R (rather than q) reduced observer variability. We conclude that for point measurements, for systolic time intervals, pulse transmission time, and RR interval, recording speed between 50 and 200 mm/s showed no statistical differences, though smallest numerical differences occurred at 100 mm/s. For LVET, 25 mm/s was satisfactory.


Observations on intravenous administration of lignocaine in patients with myocardial infarction. Lignocaine was administered intravenously to 36 patients with acute myocardial infarction. A bolus of 100 mg followed by an infusion of 2 mg/minute failed to maintain plasma levels above 2 microgram/ml. A bolus of 100 mg followed by 4 mg/minute also failed to maintain satisfactory plasma concentrations during the first hour of therapy. A bolus of 75 mg was combined with an infusion of 10 mg/minute for 20 minutes followed by 1.5 mg/minute. Satisfactory plasma concentrations during the first hour were observed in 94 per cent of the estimations. No important adverse side effects occurred during the infusion of 10 mg/minute.


Atypical Wenckebach periodicity simulating Mobitz II AV block. Eleven patients were studied and a total of 144 Wenckebach cycles in the AV node and 118 Wenckebach cycles in the His-Purkinje system were analysed to determine the incidence of typical and atypical Wenckebach periodicity, with particular emphasis on one variant of atypical Wenckebach that may simulate a Mobitz type II block. This pseudo-Mobitz II pattern was defined as a long Wenckebach cycle in which, at least, the last three beats of the cycle show relatively constant PR intervals (variation of no more than 0.02 s in surface leads and no more than 10 ms in His bundle electrograms) and in which the PR interval immediately following the blocked beat is shorter than the PR interval before the block by 0.04 s or more. Atypical Wenckebach cycles were found to be more common than the typical variety at both the AV node (67%) and His-Purkinje system (69%). The pseudo-Mobitz II pattern was seen in 19 per cent of atypical AV nodal Wenckebach periods and in 17 per cent of atypical His-Purkinje system Wenckebach cycles. The need to discern a 'classical' Mobitz II block from a pseudo-Mobitz II pattern, especially in the setting of an acute inferior myocardial infarction, is emphasised.


Histopathological correlates of sinoatrial disease. Six hearts from patients suffering from rhythm disorders consistent with the diagnosis of sinoatrial disease were histologically examined. Four of the patients had shown a tachycardia-bradycardia syndrome, and the remaining two patients episodes of sinus arrest or sinoatrial block with a slow junctional escape rhythm. The rhythm disorders had occurred in the setting of chronic sinoatrial disease (3 cases), acute myocardial infarction (2 cases), and diphtheritic myocarditis (1 case). The abnormalities which were more consistently observed consisted of (1) total or subtotal destruction of the sinus node (6 cases); (2) total or subtotal destruction of the areas of nodal atrial continuity (5 cases); (3) inflammatory or degenerative changes of the nerves and ganglia surrounding the node (6 cases); (4) pathological changes in the atrial wall (5 cases). Chronic or acute lesions involving the AV node, the bundle of His, and its branches or their distal subdivisions were also found in all 6 hearts. The relationship between the observed pathological changes and the physiological disorders are discussed.


Continuous electrode monitoring of systolic time intervals during exercise. Current systolic time interval techniques have limited clinical applicability since patient co-operation and attention to the carotid pulse and phonocardiogram transducers are required. Therefore only surface electrodes were used to monitor the electrocardiogram and electrical impedance cardiogram first derivative (dZ/dt) in the acquisition of the timing signals. dZ/dt motion artefacts were eliminated by computerised ensemble averaging, thus permitting uninterrupted data acquisition. We studied the continuous response of multistage treadmill exercise on 13 normal volunteers, since maximal distortion of noninvasive measurements occurs in dynamic exercise. The individual response trends were combined for 6 symbolic indices and each mean index had a high statistical significance (P less than 0.001). This new method surveys continuously ventricular performance with surface electrodes and therefore has the potential of monitoring the ventricular performance of critically ill patients.


Effects of prostaglandin E1 on pulmonary circulation in patients with pulmonary hypertension. The effects of prostaglandin E1 on pulmonary circulation and left ventricular performance have been studied in 20 patients with mitral valve disease and pulmonary hypertension. Prostaglandin E1 was administered intravenously over a period of 30 minutes. The dose used was 0.01 microgram/kg per min during the first 15 minutes and 0.02 microgram/kg per min subsequently. The first dose led only to an insignificant fall in left ventricular end-diastolic pressure. Infusion of prostaglandin E1 in a dose of 0.02 microgram/kg per min resulted in a significant fall in the pulmonary arterial pressure (P less than 0.001), total pulmonary resistance (P less than 0.001), left ventricular end-diastolic pressure (P less than 0.001), and aortic pressure (P less than 0.01), and an increase in the pulmonary blood volume (P less than 0.01), cardiac index (P less than 0.01), and heart rate (P less than 0.05). No significant differences were noted in stroke volume index or left ventricular dP/dt at 50 mmHg after prostaglandin E1. These results indicate that exogenously administered prostaglandin E1 causes active vasodilatation of the pulmonary vascular bed and has no inotropic action on the cardiac muscle.


Congenitally bicuspid aortic valves. Clinicogenetic study of 41 families. The families of 41 patients with surgically proved isolated bicuspid aortic valves were examined. There were 275 first degree relatives of whom 220 were living, and 188 (85.5%) of these were examined. Seven first degree relatives were found to have aortic valve disease, and in a further 11 there was 'doubtful' evidence of bicuspid aortic valves. In 6 families there was more than 1 affected member and in an additional 7 families there was 1 or more 'doubtful' first degree relative. The minimum family incidence was therefore 14.6 per cent, or 31.7 per cent if 'doubtful' cases were included. The inheritance is most probably multifactorial, but occasionally the condition may occur as an autosomal dominant. The difficulties of diagnosing bicuspid aortic valves before the development of obstruction of left ventricular outflow were encountered and are discussed. The association of a bicuspid aortic valve with asymmetric septal hypertrophy, hypertrophic cardiomyopathy, and Marfan's syndrome was also noted.


Spontaneous coronary artery perforation with tamponade. Demonstration by necropsy selective coronary arteriography. Pericardial tamponade in a 51-year-old man after acute anteroseptal myocardial infarction was the result of spontaneous perforation of an atheromatous coronary artery. The value of a simple necropsy technique of selective coronary arteriography in demonstrating the lesion is described.


A case of rhabdomyosarcoma of the right ventricle. A patient is described who presented with symptoms suggesting pericarditis and cardiac tamponade, and who was found at necropsy to have a rhabdomyosarcoma of the right ventricle. The presenting features of right ventricular tumours are discussed and some useful pointers to the correct diagnosis suggested.


The significance of vertical mitosis in epidermis. In normal epidermis mitosis usually occurs in a plane horizontal to the dermo-epidermal junction, both daughter cells then remaining basal while an adjacent non-mitotic cell is detached from the baseline and forced distally. The evidence now indicates that only cells in a certain intermitotic state, called GIb, can be thus detached, and that when such cells are not locally available the expanding mitosis is forced into the vertical axis. The higher the mitotic rate the shorter is the intermitotic interval, and therefore the less probable it becomes that GIb cells will be locally available for extrusion and the more probable that mitosis will take place vertically. At the highest mitotic rates, as in hair bulbs, all mitoses are vertical. The relation of these findings to epidermal structure and to epidermal cellular homeostasis is discussed.


Dermatoglyphics in dermatitis herpetiformis. In a dermatoglyphic study of 101 patients with dermatitis herpetiformis, comparison was made with their normal relatives and several random normal series. The results of the several comparisons are quite consistent, considering the nature of the data, for the all point to an attenuation of qualitative and quantitative digital and palmar traits in the patients. It is argued that these differences are associated with the disease itself, and that some of the female relatives have an inherited tendency to the disorder, but do not express it. The findings suggest the involvement of genetic factors in the aetiology, and possibly intrauterine environmental influences as well.


Topical betamethasone 17-valerate is an anticorticosteroid in the rat. 1. Dermal atrophy. In the albino rat, topical betamethasone 17-valerate acts as an anticorticosteroid. This steroid is inactive in a dermal atrophy assay over a dose range where betamethasone and hydrocortisone display atrophogenic activity. At appropriate concentrations betamethasone 17-valerate competitively inhibits the atrophogenic effects of both betamethasone and triamcinolone acetonide. Since betamethasone and betamethasone 17-valerate penetrate rat skin in vivo at essentially the same rate, it is concluded that the latter compound is relatively resistant to hydrolysis during penetration, and that it binds to rat corticosteroid receptor proteins in such a manner as to prevent expression of corticosteroid activity. Therefore, the rat cannot be used as a model species to predict activity in man for this compound.


Topical betamethasone 17-valerate is an anticorticosteroid in the rat. 2. Anti-inflammatory and anti-lymphocyte activities. In the oxazolone-induced delayed hypersensitivity inflammation in the rat ear, betamethasone 17-valerate, in contrast to other topical corticosteroids, is incapable of suppressing oedema. When given in combination with triamcinolone acetonide, betamethasone 17-valerate competitively antagonizes the anti-inflammatory action of the active steroid. When tested in the mouse, betamethasone 17-valerate behaved as an anti-inflammatory agent 15 and 80 times as potent as betamethasone and hydrocortisone respectively. In an in vivo lymphocyte culture system in which preincubation with corticosteroids prevents subsequent phytohaemagglutinin induced DNA synthesis, betamethasone 17-valerate was less active than even hydrocortisone when rat lymph node cells were used, but with human cell preparations it was more potent than either hydrocortisone or betamethasone. Betamethasone 17-valerate behaves uniquely in the rat as an anticorticosteroid; in mouse and in man the compound behaves as a normal corticosteroid.


Collagen sponge for leg ulcers.

Collagen sponge for leg ulcers. Collagen sponge, reconstituted from bovine tendon, provides a porous matrix for the permeation of fibroblasts and capillaries and on theoretical grounds it should be ideal for stimulating granulation tissue in deep leg ulcers. A preliminary study in six patients has suggested that a collagen sponge dressin enhanced granulation, but subsequent re-epithelialization over the reconstituted collagen tended to be slower than usual.


Induction of mixed-function oxidases in mouse liver by psoralens. 8-methoxypsoralen, the most widely used psoralen in photochemotherapy, was shown to induce mixed-function oxidases. Induction of mixed-function oxidases in mouse liver was examined after mice were given a single dose of one of three psoralens. 8-methoxypsoralen clearly induced p-nitroanisole O-demethylase and slightly induced aryl hydrocarbon hydroxylase. Psoralen and 4,5',8-trimethylpsoralen failed to induce either enzyme. These results represent the first enzyme induction studies on these clinically useful compounds. The studies may have relevance in understanding the clinical differences in skin photosensitizing activity and photochemotherapeutic effectiveness of the compounds in such diseases as psoriasis and vitiligo.


Evaluation of PUVA, topical corticosteroids and the combination of both in the treatment of psoriasis. A total of 90 patients with psoriasis were treated either by photochemotherapy (PUVA) or by topical steroids under occlusion (TOC). In a third group of patients a combination of both regimens was applied. All patients received approximately 15 treatments of either type. During therapy, biopsies were taken in all treatment groups and the epidermal thickness was monitored planimetrically. After clearing of the skin the patients were left without treatment and the time of onset of new lesions was recorded. While TOC produced rapid clearing of the skin this was followed by early relapses (in 50% of the patients after 3 weeks).  PUVA reduced psoriatic hyperplasia more slowly; however, 50% of the patients remained free of psoriasis for approximately 10 weeks after clearance. Psoriasis treated with steroids under occlusion together with PUVA showed a significantly faster return of the skin to normal as compared to PUVA. As in PUVA-treated patients relapses occurred after approximately 12 weeks is this group. Therefore the combination of both treatments appears to be advantageous for rapid skin clearance and comparatively long remissions.


Trioxsalen bath plus UVA effective and safe in the treatment of psoriasis. Seventy-four patients with psoriasis were treated using a trioxsalen bath (50 mg/150 l of water) and long wave ultraviolet light (UVA) given in an ordinary PUVA-cabin. Good or excellent results were observed in 92% of the patients in the initial phase and in 63% during the maintenance treatment. Because of local side-effects the therapy was discontinued in two patients. One of them developed contact hypersensitivity to trioxsalen and the other developed blisters with such low doses of UVA that it was difficult to maintain the proper dose. The therapy was started with 0.28 J/cm2 of UVA and after an average of 18 treatments, when the average dose was 1.70 J/cm2, the patients were moved to maintenance treatment which took place at 1--4 week intervals. The therapy was well tolerated and cosmetically very acceptable. The final tan was even on all but the face, which remained untanned.


TruCutR needle biopsy in asbestosis and silicosis: correlation of histological changes with radiographic changes and pulmonary function in 41 patients. A percutaneous needle biopsy was performed with a TruCut needle on 41 patients with suspected pneumoconiosis. Patients selected for biopsy tended to have brief or unusual dust exposure, as well as questionable radiographic opacities. Sixteen had been exposed to asbestos, 13 to silica and 12 to mixed dust containing quartz, coal, iron, asbestos and talc. All patients in the asbestos group and most in the other two groups had a reduced transfer factor. Most patients in the asbestos group and about 25% of the other patients had restrictive ventilatory impairment. Chest radiographs were assessed according to standard films of the ILO U/C International Classification (International Labour Office, 1972). In 25 patients radiographic opacities were absent or acanty (categories 0--1/1). The dominant radiographic feature of many patients exposed to asbestos was a ground-glass appearance or a bilateral elevation of the diaphragm, or both, features difficult to assess according to the ILO U/C scheme. Most histological changes were those usually seen in pneumoconiosis. However, in only two patients with silicosis were silicotic nodules detected. The specimens of seven patients showed a granulomatous inflammation. The severity of alveolar wall involvement correlated well with the transfer factor value but poorly with radiographic changes. The profusion of radiographic opacities also correlated poorly with functional impairment. As a diagnostic tool the needle biopsy was valuable in asbestosis and slightly less so in mixed-dust fibrosis. The biopsy specimens showed changes compatible with asbestosis in 75% of the suspected cases and in 86% of those in which asbestosis was the final diagnosis. In the mixed-dust group pneumoconiosis was confirmed in 67% and 80%, respectively. In the diagnosis of silicosis an open biopsy is probably more reliable than a percutaneous one, particularly if radiographic changes are minimal. Histological changes in the needle biopsy specimen were compatible with silicosis in only 36% of the suspected cases and in 63% of those in which the final diagnosis was silicosis.


The effect of acrylamide on human polymorphonuclear neutrophils in vitro. Acrylamide (CH2CHCONH2), the vinyl monomer of the industrially useful polymer polyacrylamide, is a recognised neurotoxin. Investigation in our laboratory indicated that, in addition to its neurotoxic effect, acrylamide depressed human polymorphonuclear leucocyte (PMN) chemotaxis in vitro. As genetic or chemical inhibition of PMN phagocytic function frequently pre-disposes patients to repeated bacterila infections, the in vitro effects of acrylamide on several other human PMN functions were studied. Acrylamide in concentrations up to 37.5 mg/ml had no effect on trypan blue uptake. However, bacterial ingestion, killing, and induced chemiluminescence were depressed by pre-treatment with acrylamide (10 mg/ml). It seems unlikely that acrylamide exposure alters host resistance to bacterial infections, because (a) large doses of acrylamide are necessary to interfere with phagocytic functions, (b) acrylamide reacts readily with proteins on many tissue cells and may be made inaccessible or non-toxic to PMN'S and (C) PMNs have a rapid turnover rate in the body and non-functional cells would be rapidly replaced by functional cells.


The effects of water restriction and water loading on urinary excretion of lead, delta-aminolevulinic acid and coproporphyrin. Alterations in daily urinary excretion of lead, delta-aminolevulinic acid (ALA), coproporphyrin, creatinine and total solutes following water restriction and water loading were examined in nine lead workers. Excretion of lead, ALA and total solutes was significantly decreased when urinary volume was reduced, showing that these values are dependent on urinary volume: conversely, excretion of coproporphyrin and creatinine was independent of urinary volume. Excretion of lead and total solutes was also dependent on creatinine excretion. The renal excretory mechanism of lead, ALA and coproporphyrin is discussed in the light of these findings.


Blood styrene and urinary metabolites in styrene polymerisation. The results of the analysis of blood and urine samples for styrene and its metabolites in 491 workers in a styrene polymerisation plant in the United States are reported. The levels of exposure to styrene were estimated to be less than 10 ppm, but nevertheless styrene and metabolites were detectable in more than 50% of workers in polymerisation jobs, within 4 h of exposure. Workers involved in the manufacture and purification of styrene from ethyl benzene also had detectable blood styrene and urinary metabolites in 83% of recently exposed subjects. The relationship between styrene in blood and in subcutaneous fat and urinary metabolites as pharmacokinetic variables is discussed.


Urinary hippuric acid concentration after occupational exposure to toluene. The results of industrial investigations have shown a correlation between the rate of hippuric acid excretion in a single urine sample collected after daily occupational exposure and the amount of toluene absorbed. The rate of hippuric acid excretion and the average concentration of toluene vapour during exposure time were also related. The quantitative range of the test has been limited to amounts exceeding 425 mg of toluene and concentrations exceeding 69 ppm of toluene in the air because of the physiological presence of hippuric acid in urine. The rate of hippuric acid excretion in urine depends on diuresis and is constant for urinary fractions with diuresis of 30 ml/h. The physiological excretion rate was 20 mg/h with a standard deviation +/- 4.3 mg/h, maximal physiological level 33 mg/h.


Mourning and the prevention of melancholia. Freud suggested that a high level of ambivalence in the previous relationship between the bereaved and the deceased was likely to complicate the process of mourning, perhaps leading to melancholia. Within a study testing the effectiveness of preventive intervention, a subgroup of recently bereaved widows (n = 22) were rated as having had a highly ambivalent marital relationship with their spouses. Psychotherapeutic intervention to promote the resolution of the ambivalence within the mourning process was given to a randomly selected group of these women during the first three months following the husbands' deaths. All subjects were followed up independently by questionnaire 13 months after the death. Intervention subjects (n = 12) showed significantly better general health and a lower level of depressive illness than controls (n = 10).


The reconstruction of childhood in psychoanalysis. Reconstructions are defined as genetic propositions showing not only what happened to the individual in childhood, but also how he adopted his particular solutions to conflict situations in the past and how he continues to be guided today by that experience of the past. The relative neglect of environmental failures in psychoanalysis is traced to Freud's shift from physical to psychic reality, although the gains from this shift are acknowledged. The discussion of fantasy and historical fact proceeds to an examination of the case for and the case against the use of reconstructions, and the nature of memory in psychoanalysis. A clinical illustration leads to a synthesis of the conflicting views, emphasizing that both the analyst and the patient need to remain in touch with the past, the current life, and the transference relationship. The paper is concluded with the views of modern historians.


Psychological construing and thought disorder: another test of the 'difficulty' hypothesis. Two versions of the grid test of thought disorder were derived which used similar elements, and were of equal difficulty, but one of which employed psychological, and the other non-psychological, constructs. They were administered to thought-disordered schizophrenic patients and to psychiatric and normal controls. On one measure (consistency), although not on the other (intensity), the results strongly supported the hypothesis that thought-disordered patients are maximally disordered in the area of psychological construing. The results support those of an earlier study (McPherson et al. 1975) and are compatible with those theories which postulate a role for social factors in the aetiology of thought disorder.


Objective constructs and cognitive structure. Some relations between objective content and concrete structure in cognitive systems are examined in repertory grids completed by 58 individuals. Predictions that systems with objective content would show less discrimination, differentiation and integration were unconfirmed, results tending in the opposite direction to that hypothesized. There was a similar outcome for six measures of cognitive style as dependent variables. It it suggested that objective-subjective constructs and their concrete-abstract use should be further investigated with the expectation of finding more complex relations in individuals' cognitive systems.


Contingency learning in chronic schizophrenia and its relevance to social motivation deficit. The present study investigates the ability of chronic schizophrenic patients to learn to obtain social rewards when the incentive value of the contingent social event is known. From a group of 40 chronic patients tested for responsivity to social rewards, socially responsive and socially unresponsive patients were selected and compared on a learning task. Patients who were highly motivated to obtain social reinforcement did not emit the reinforced response during 300 learning trials any more frequently than did patients who were not motivated by social rewards. It was only when the experimental contingency was specified that responsive and unresponsive patients could be differentiated. The implications of these findings for social motivation theories of schizophrenia are discussed.


The hysterical personality--I. The healthy hysteric. This paper examines the meaning and validity of 'hysterical personality disorders'. Theories and descriptions of hysterical personalities have been conflicting and often mutually incompatible, leading to a vague and imprecise nosology. No generally accepted specific inclusion criteria or follow-up studies are available; consequently, the diagnosis of 'hysterical personality disorder' has reflected adaptive as well as maladaptive traits. This paper critically examines the healthy dimension of Lazare's continuum of personality pathologies exhibiting hysterical traits. The continuum, in an attempt to reconcile the inconsistencies of this diagnosis, ranges from the relatively healthy (good, genital, true) hysteric, to the sick (bad, oral, 'so-called good') hysteric. But this paper concludes that here, too, evidence for diagnostic legitimacy is lacking. Women at the healthy end of the hysterical personality spectrum are best regarded as healthy and do not meet criteria for the diagnosis of a personality disorder.


Cervical cancer and cytology screening in New Zealand. Cytology screening, used in New Zealand since 1955 at an intensity comparable to that in Canada generally, has not favourably affected incidence and mortality rates for cervical cancer; these have actually risen significantly in 20 to 34-year-old New Zealand women. Canadian claims that mortality falls are related to intensity of cytological screening are not justifiable, so that the significance of the 'pre-cancers' revealed by cytology and the value of population screening would seem to be doubtful.


The pattern of presentation of carcinoma of the uterine cervix in East Anglia 1960 to 1975. Changes in the pattern of presentation of carcinoma of the uterine cervix between 1960 and 1975 were studied using the records of 3193 patients registered with the East Anglian Cancer Registration Bureau. Between 1963 and 1968, an increased registration rate for invasive tumours was largely attributable to increased registrations of Stage I and II lesions in the 35 to 49 age group. After 1970 the registration rate for lesions of all stages in this age group declined. Over the same period the 50 to 64 age group showed a tendency for an increased proportion of registrations to be of Stage I or II lesions. There was little change in pattern in the over-65 age group. The possible relation of these changes to the introduction of a cervical cytology screening programme and the implications for future screening policies are discussed.


Distinction of benign from malignant ovarian cysts by ultrasound. The ultrasonic characteristics of benign and malignant cystic ovarian tumours have been reviewed. The value of each of eight different ultrasonic features has been assessed and the specificity of each for confirmation or exclusion of malignancy is indicated. In combination they permit correct differentiation in 91 percent of cases.


Significance of antepartum cardiotocography in normal pregnancy. Antepartum cardiotocographs (CTGs) were obtained at weekly intervals from 34 weeks onwards and twice weekly from 40 weeks onwards in 91 strictly normal pregnancies. The Fischer score was used to assess the 594 CTGs obtained. Ominous patterns were not encountered but 7.2 per cent of CTGs showed a suspicious pattern. Absence of accelerations and an oscillatory amplitude of less than 10 beats/minute contributed to respectively 95 and 88 per cent of these patterns. The combination of reduced variability and absence of accelerations accounted for 84 per cent of suspicious CTGs. This pattern is compatible with normal pregnancy even when fetal movements are present. One-third (37 per cent) of perfectly normal pregnancies had a suspicious CTG on at least one occasion and in 9 per cent of cases it was found twice or more. This should be borne in mind when using antepartum CTG in the surveillance of high-risk pregnancy.


Secretion patterns of plasma-progesterone, 17-hydroxyprogesterone and 20 alpha hydroxypregn-4-en-3-one in early normal pregnancy. Plasma levels of progesterone, 17-hydroxyprogesterone and 20 alpha hydroxypregn-4-en-3-one were established from 4 to 16 weeks gestation by sampling 15 normal patients twice weekly. Apart from a peak at week 5, the mean levels of progesterone remained constant at about 24 ng/ml until week 10 and then rose progressively to 40 ng/ml at week 16.20 alpha hydroxypregn-4-en-3-one remained within the 7 to 9 ng/ml range apart from a peak at week 5. A significant decrease in mean levels of 17-hydroxy-progesterone was noted from week 5 to a nadir of 3 ng/ml at week 11. It would appear that the main hormone production of progesterones in pregnancy has been taken over by the placental unit by the 10th week.


Secretion patterns of plasma-progesterone, 17-hydroxyprogesterone, and 20alpha hydroxypregn-4-en-3-one in early abnormal pregnancy. Plasma progesterone, 17-hydroxyprogesterone, 20alpha hydroxypregn-4-en-3-one levels were determined twice weekly up to 16 weeks gestation, where possible, in a twin pregnancy, in two patients who aborted spontaneously and in three patients who were treated with 'progesterone supplements' because of abnormal vaginal cytology. There was no correlation between vaginal smears and the plasms hormone levels and there was no evidence to suggest that progesterone supplements influenced clinical outcome. Compared with normal mean values the only difference was a significantly rise in progesterone and 20alpha hydroxypregn-4-en-3-one levels in the twin pregnancy after the 12th week and a precipitate fall in all hormone levels just prior to abortion. Plasma hormone levels could not be used to predict outcome.


Specular microscopy of the corneal endothelium. The endothelium of the normal corneas of 67 human subjects was studied in vivo with the specular microscope in order to quantify the method as a means of sampling the cell density of the tissue. It was found that (1) axial cell counts of the endothelium are reproducible in the same cornea after an interval of time; (2) the cell counts of the centre and periphery of the same cornea are similar; (3) the axial cell counts of pairs of eyes are similar; and (4) there is a gradual reduction of cell number with increasing age. The significance of these data is discussed.


Effects of intraocular miotics on cultured bovine corneal endothelium. Two cases of severe corneal oedema occurred after the use of intraocular pilocarpine. Experimental investigations were conducted with cultured bovine corneal endothelial cells exposed for 5 minutes to 1% pilocarpine solutions of varying composition. Cells were destroyed in solutions not isotonic with aqueous humour, and calcium-free ionic solutions caused loss of cell adhesion without loss of viability. Low pH or the presence of 1% pilocarpine had no detectable effects; 1% acetylcholine chloride in 5% mannitol (Miochol) also caused cell destruction, and this preparation was found to be considerably hypertonic. The minimum requirements for the formulation of intraocular miotics are discussed.


The reverse Wessely phenomenon: immune corneal rings following systemic immunisation. Rabbits were hyperimmunised systemically over a 4-week period with bovine serum albumin (BSA) in complete Freund's adjuvants and were subsequently challenged systemically with a massive dose of BSA designed to produce serum sickness. Without any ocular manipulation a spontaneous corneal immune reaction was observed which we termed a 'reverse Wessely phenomenon'. It was due to a reversed sequence of antibody becoming sequestered in the cornea and later responding to antigen entering via the limbal circulation. In the stroma, lymphocytes were prominent and only few polymorphonuclear cells were found, but this may have been due to late sectioning of the corneas tested. The corneal ring formed at the limbus and migrated centripetally ahead of peripheral clouding, which is typical of the Wessely phenomenon. Possible relationships of this reaction with some autoimmune disorders are discussed.


Scanning electron microscope study of herpes simplex virus experimental disciform keratitis. Scanning electron microscopy of the endothelium of experimental disciform keratitis revealed corneal endothelial changes which distinguished disciform oedema from the more progressive stages of disciform keratitis. The endothelium of corneas with disciform oedema were wholly intact and characterised by subtle morphological alterations. In contrast, the more progressive stages of disciform keratitis were characterised by massive destruction and denudation of the endothelium. The significance of these observations is discussed.


Acute follicular conjunctivitis and keratoconjunctivitis due to herpes simplex virus in London. During the 18 months January 1975 to June 1976, 25 cases of acute herpetic follicular conjunctivitis and keratoconjunctivitis resembling adenovirus ocular infection presented in the External Eye Disease Clinic, Moorfields Eye Hospital, City Road, London. Herpes simplex virus was isolated in HEp2 cells in 22 patients, and the remaining 3 patients were identified by a minimum 4-fold rise in the level of antiherpes simplex virus antibody in their blood. No adenovirus was isolated from these patients, but complement fixation test for adenovirus was positive in 1 patient with cultural test positive for herpes simplex virus. Most patients were between 20 and 35 years old and the ratio of males to females was 12 to 13. At the initial visit the clinical features of disease were moderate to severe conjunctival papillary and follicular reasons with epithelial and subepithelial punctate keratitis but little systemic disease. In the absence of typical herpetic lesions of face, lids, or cornea the disease resembled adenovirus types 8 or 19 keratoconjunctivitis. Of these 25 patients 5 subsequently developed typical herpetic lesions of lids or cornea. In the remaining 20 cases the correct diagnosis could be made only by cultural or serological tests. Virological diagnosis provides a rational basis for antiherpetic chemotherapy, which appears to shorten the course of infection.


Studies on the effect of the lipid phase transition on the interaction of glucagon with dimyristoyl glycerophosphocholine. Glucagon is found to interact with dimyristoyl glycerophosphocholine both above and below the phase transition temperature of the lipid. Above the phase transition temperature the interaction is manifested by an increase in the rate of vesicle aggregation and by an increased permeability of unilamellar vesicles to Eu3+ and to Fe(CN)3-6. However, no stable lipoprotein complex can be detected by gel filtration. Below the phase transition glucagon can form stable complexes with dimyristoyl glycerophosphocholine vesicles but cannot rapidly  rearrange these vesicles to disk-shaped particles until the phase transition temperature is approached. The energy of activation for the dissociation of glucagon from the disk-shaped lipoprotein particle is 29 kcal/mol at temperatures above 36 degrees C but increases markedly at lower temperatures, as the region of the lipid phase transition is approached. This increase in energy of activation at lower temperatures is most probably due to the larger amount of energy required to rearrange gel-state lipid in the transition state and provides an explanation for the unusual kinetic stability of the glucagon-dimyristoyl glycerophosphocholine lipoprotein complex only at temperatures below the phase transition of the lipid.


The Arrhenius plot behaviour of rat liver 5'-nucleotidase in different lipid environments. The plasma membrane enzyme 5'-nucleotidase (EC 3.1.3.5) was prepared from rat liver as a complex with sphingomyelin or in detergent-solubilized forms. Each preparation exhibited a Km and Arrhenius break temperature indistinguishable from that of 5'-nucleotidase in plasma membranes. Measurement of fatty acid profiles, cholesterol and phospholipid content however showed a very wide variation between these preparations. We conclude that the biphasic nature of the Arrhenius plot of 5'-nucleotidase may be a property of the enzyme rather than its lipid environment.


The relationship between calcium flux and beating in hyperpermeable heart muscle cells. Beating hyperpermeable myocardial fragments, suspended in fresh medium, show a Ca uptake rate which greatly exceeds the release rate. Under these conditions, Ca influx and contraction frequency co-vary with temperature. Ca efflux, however, is not temperature responsive. In addition, influx, but not efflux, is ATP-dependent. This implies that influx, but not efflux, is a controlled process, the activity of which has the same determinants as does contraction. However, under conditions approximating the steady state, the Ca influx rate was found to decrease, while the efflux rate increased. This suggests the possibility, at least, that the source of contraction-activating Ca can vary with the prevailing conditions.


Chemical characterization of glomerular and tubular basement membranes of cattle of different ages. Glomerular and tubular basement membranes were isolated from fetal, neonatal, young and adult bovine kidneys. An isolation method with sieves for both glomeruli and tubules from the same kidney was developed. A detergent procedure appeared to give purer glomerular and tubular basement membrane preparations than the generally used sonication method. No large differences were found in the composition of glomerular and tubular basement membrane of adult animals. Glomerular and tubular basement membrane preparations of the four age groups showed an increase with age of hydroxylysine and both 3- and 4-hydroxyproline. The most marked increases appeared at different stages of development, that of tubular basement membrane being between fetal and neonatal stages and glomerular basement membrane between 18 weeks old and adult animals. The ratio of 3- to 4-hydroxyproline increased considerably during development. Total imino acid content was higher for both types of basement membrane from adult than from young animals, while total content of hydroxylysine plus lysine remained fairly constant. The increase in hydroxylation of lysine was accompanied by a corresponding change in glucose and galactose content so that the ratio of galactose to hydroxylysine or glucose to galactose remained constant. Fucose content of both types of basement membranes was the same for all age groups but content of aminosugars and mannose gradually increased with age.


The structure of oriented sphingomyelin bilayers. X-ray diffraction from oriented bilayers of sphingomyelin gave up to 14 orders of diffraction of a lamellar repeat of 68.5 A on the merididan and up to eight reflections, including a strong reflection at 4.2 A, on the equator. The diffraction spacings did not change when the sphingomyelin bilayers were exposed to different humidities. A direct analysis of the low resolution X-ray data, using deconvolution is presented. A comparison of the Patterson functions of sphingomyelin with those of phosphatidylcholine and phosphatidylethanolamine suggests that the molecular structure of sphingomyelin in oriented bilayers resembles the structure of both phosphatidylcholine and phosphatidylethanolamine. Molecular model calculations for sphingomyelin bilayers have also been performed. Electron density profiles of sphingomyelin bilayers at resolution of about 6 A and about 2.5 A are presented. Our results indicate that the phosphorylcholine head group of sphingomyelin is in the plane of the membrane and at right angles to the hydrocarbon chains, the hydrocarbon chains are nearly parallel to each other, and there is only a limited, if any, interdigitation of the hydrocarbon chains of the adjacent sphingomyelin molecules in the bilayer.


Calcium-induced aggregation and fusion of mixed phosphatidylcholine-phosphatidic acid vesicles as studied by 31P NMR. 1. The transbilayer distribution of the phospholipids in sonicated egg phosphatidylcholine-phosphatidic acid vesicles and the interaction of Ca2+ with these vesicles was studied by 31P NMR. 2. Over a wide composition range the bilayer of these vesicles has a symmetrical phospholipid composition. 3. With ratios of Ca2+ to phosphatidic acid in the outer monolayer of the vesicles up to 0.3, Ca2+ induces vesicle aggregation. The extent of aggregation is increased by the Ca2+ concentration in the medium and the outer monolayer concentration of phosphatidic acid. The vesicle aggregation can be fully reversed by chelating Ca2+. 4. When the ratio exceeds 0.5 Ca2+ induces vesicle fusion. The fusion is maximal for vesicles containing both phosphatidylcholine and phosphatidic acid. The data suggest that Ca2+-induced lateral phase separations make the bilayer more susceptible to fusion.


Laser Raman studies of molecular interactions with phosphatidylcholine multilayers. II. Effects of mono- and divalent ions on bilayer structure. Laser Raman spectroscopy was applied to characterize structural behavior of dipalmitoyl phosphatidylcholine multibilayer systems in the presence of several cations (K+, Na+, Cs+, Rb+, Ca2+, Mg2+, Cd2+, Ba2+) and anions Cl-, Br-, I-, NO-3, SO2-3, SO2-4, S2O2-3, S2O2-8). To evaluate the Raman-spectroscopical data quantitatively, characteristic intensity ratios, lateral and trans order parameters were used and compared. It was shown that the different trans order parameters are rather sensitive to ion-polar head group interactions and thus, they cannot give unequivocal information on the trans-gauche isomerization of hydrocarbon chains of phospholipids. The observed effects of ions on Raman spectra of phospholipid multilayers could not be explained simply on the basis of electrostatic interactions. The possible involvement of other factors (changes in polarizability, hydrogen bonds, etc.) is also discussed. It was demonstrated that the order parameters defined in different ways may result in different effectiveness sequences of ions. Of monopositive ions Na+ was found to be the most effective to influence the bilayer structure. For dipositive ions, of which Ca2+ proved to be the most effective, concentration-dependent effectiveness sequences were obtained. A plausible interpretation and some consequences of the concentration-dependent two-step binding of divalent cations were also outlined. Bilayered phospholipid structures turned out to be more responsive to anions than to most cations investigated. Interdependent actions of cations and anions, as well as the possible relevance of the charge distribution on anions are postulated.


Local osmotic coupling to the active trans-endothelial bicarbonate flux in the rabbit cornea. The present experiments investigate HCO3-, Cl- and fluid fluxes across partially destromalised corneas. Although there is no net flux of Cl-, there is a net flux of HCO-3 across the endothelium from stromal side to aqueous side which is accompanied by a flux of water in the same  direction. Bulk phase osmosis cannot account for the initiation of the flux of fluid. Local osmotic coupling between ions and water is postulated to occur in the preparation. The exudate is hypertonic to the bathing Ringer solution.


Kinetics studies on the renal transport of probenecid in vitro. 1. The kinetic parameters of renal transport of probenecid have been assessed by studying the uptake of the drug in rabbit kidney tubules incubated in an electrolyte medium under various conditions. 2. The added compounds inhibited the uptake of probenecid both by kidney cortical slices and separated renal tubule preparations in the following order: p-aminohippurate less than phenol red less than bromophenol blue less than bromocresol green. A reversible competitive inhibitory effect of these organic anions on the renal accumulation of the drug was observed. 3. The Km for renal uptake of probenecid in separated tubules (0.04 mM) and the KI values calculated in this system for p-aminohippurate (0.5 mM), phenol red (0.09 mM), bromophenol blue (0.02 mM) and bromocresol green (0.015 mM) were found to be in good agreement with the corresponding KI value of probenecid and Km values of these compounds previously observed in various kidney tissue preparations. 4. On the basis of above mentioned findings, it is concluded that probenecid, p-aminohippurate and various phenolsulphonphthalein dyes are transported by the common renal organic anion transport system.


Protein synthesis on membrane-bound and free ribosomes in growing yeast. 1. A method for the study of membrane-binding of ribosomes and the rate of synthesis of protein on free and bound ribosomes has been examined critically. 'Brij 58' has been found to be more suitable for detaching ribosomes from membranes than 'Triton X-100' or sodium deoxycholate since it does not alter the structure or biological activity of the ribosomes. Triton and deoxycholate remove amino acid- and ribonucleotide-containing material from ribosomes, thus decreasing their activity during protein synthesis in a cell free system. 2. The rate of protein synthesis on membrane-bound and free ribosomes, the total amount of ribosomes, and the percentage distribution of free and membrane-bound ribosomes were determined in a culture of synchronously growing yeast cells. The results show that in all probability protein synthesis in yeast cells occurs mainly on membrane-bound ribosomes. Free ribosomes are considered here to be a ribosome pool which allows the cell to increase its rate of protein synthesis rapidly. Membrane-bound and free ribosomes are viewed as different functional states, one being a state of protein synthesis and the other a latent state.


Proteins bound to poly(A) sequences of polyribosomes from germinating wheat embryos. A preparation of purified polyribosomes was isolated from wheat embryos germinating for 1 h with [3H]adenosine. Treatment of polyribosomes with pancreatic ribonuclease leads to the liberation of particles containing poly(A) sequences. Poly(A)-containing complexes show a heterogeneous distribution in the region 8--15 S. The ability of 8--15 S material to be absorbed to membrane filters under the conditions of low ionic strength and its sensitivity to pronase indicate that poly(A) sequences in polyribosomes are associated with protein. Analysis of this protein by electrophoresis in a polyacrylamide gel demonstrates the presence of two polypeptides with the molecular weights equal to 86,000 and 52,000.


Effect of ethidium bromide on the digestion of chromatin DNA with micrococcal nuclease. Intercalation of ethidium bromide into DNA influences the rate of its digestion with micrococcal nuclease in opposite directions depending on whether it is free DNA or DNA in chromatin. In the case of free DNA the binding of ethidium bromide, starting from a very low concentration, results in the inhibition of the rate of digestion (increasing constantly with the increase of the ethidium bromide/nucleotide ratio). In contrast to free DNA the digestion rate as well as the overall amount of nuclease susceptible DNA is increased upon ethidium bromide binding to chromatin, with maximum enhancement around the saturation of intercalation sites. The saturation of intercalation sites in chromatin leads also to the disappearance of the typical micrococcal nuclease digestion pattern of DNA upon gel electrophoresis. Instead, a random cleavage pattern is observed. These data indicate that partial unwinding of chromatin DNA by ethidium bromide results in unmasking new sites for nuclease action. Interpretation of this finding in terms of the nucleosomal structure of chromatin and the mode of ethidium bromide binding to chromatin DNA indicates that newly unmasked sites are localized within the core particle DNA.


Positioning of the A . T-rich regions in rat mitochondrial DNA by electron microscopy and analysis of the hysteresis of denaturation. Intramolecular heterogeneity in the base composition of rat mitochondrial DNA (mtDNA) was shown by a combination of an improved denaturation mapping technique using electron microscopy and analysis of high-resolution optical melting-renaturation profiles. Circular mtDNA starts to melt in one specific region and then forms loops in four other regions in random order. These five early melting regions are all located in one half of the molecule. The arrangement of the early melting regions in rat mtDNA bears a remarkable resemblance not only to those of mtDNAs from several species of Drosophila but also to those of several species of Drosophila but also to those of several plasmid DNAs and phage DNA.


The human spermatozoa genome. Analysis by DNA reassociation kinetics. The physicochemical characteristics of the human spermatozoa DNA have been analyzed by means of analytical ultracentrifugation and reassociation kinetics and compared with DNA from a somatic human cell, the leukocyte. Human spermatozoa DNA is composed of a major (86.7%) and a minor (13.3%) component with buoyant densities in neutral CsCl corresponding to 1.697 and 1.704 g/cm3, respectively. The DNA obtained from leukocytes contained only the first component. Reassociation kinetics of sheared DNA showed that the most rapidly-renatured repeated sequences consituted 12.1% of the sperm genome and only 9.2% of leukocyte genome. On the contrary single copy sequences were more abundant in leukocyte (64%) than in spermatozoa (59%) DNA. Reassociation kinetics with long DNA fragments (2000 nucleotides) were clearly different between sperm and leukocyte DNA, indicating the presence in sperm cells of a specific cluster of highly repetitive DNA. This entity may play a role in giving the spermatozoa nucleus its characteristic properties, involving supercoiling, quasi crystal structure, stability, lack of transcription, etc. and secondarily may function as an early pairing mechanism during syngamy.


Methylation of yeast tRNAPhe by enzymes from cytoplasm, chloroplasts and mitochondria of Phaseolus vulgaris. Pure yeast tRNAPhe was used as a substrate to compare the tRNA methylating activities in Phaseolus vulgaris cytoplasm, chloroplasts and mitochondria, in the presence of S-adenosyl[Me-3H]methionine. The resulting [Me-3H]-tRNAPhe was then analyzed, using the techniques of nucleotide sequence determination. Cytoplasmic and mitochondrial enzymes catalyze the methylation (into m5C) of C48 present in the extra-loop, while chloroplast enzyme preparations catalyze the modification (into m1A) of A14 present in the dihydrouridine loop of tRNAPhe.


Transcription of the non-repetitive genome in liver hypertrophy and the homology between nuclear RNA of normal and 12 H-regenerating liver. The percentage of the non-repetitive genome transcribed and the complexity of nuclear RNA were estimated in normal and 12 h-regenerating rat liver. Nuclear RNA from normal or 12 h-regenerating liver hybridizes with approximately 6.1% of non-repetitive DNA (12.2% of the single-copy genome, assuming assymetric transcription). The estimated complexity of either of these nuclear RNA populations is 7.6 . 10(10) daltons, which is approximately 7 times higher than that calculated for polysomal mRNA. Cross hybridization experiments did not show differences between the nuclear RNA populations of normal and 12 h-regenerating liver. The results indicate that liver hypertropy (without hyperplasia) may be brought about without a large increase in the proportion of the non-repetitive genome transcribed.


Biosynthesis of low molecular weight RNA in mouse myeloma cells. The low molecular weight RNAs in the nucleus and cytoplasm of mouse myeloma cells have been characterized. There are major nuclear species (other than 5-S and tRNA), four of which contain 'capped' 5' termini. There are three major small cytoplasmic RNAs none of which contain 'caps'. The biosynthesis of the nuclear species when studied using [3H]adenosine as an RNA precursor is characterized by a rapid, transient appearance in the cytoplasm, followed by fixation in the nucleus. Within 15 min, the amount in the cytoplasm has reached a steady-state level maintained for 60 min, while the accumulation in nuclei is linear after a short lag (less than 5 min). When biosynthesis is studied using [Me-3H]methionine as a precursor, much less labeled RNA is present in the cytoplasm, suggesting that methylation may immediately precede fixation into the nucleus.


Synthesis and properties of O6-methyldeoxyguanylic acid and its copolymers with deoxycytidylic acid. This paper describes the synthesis of O6-methyldeoxyguanosine triphosphate (m6dGTP) and its copolymerization to high molecular weight polymer with deoxycytidylic acid. The monomer, m6dGTP, was synthesized from deoxyguanosine first protected by acetylation of the sugar hydroxyls, and then chlorinated in the 6-position with POCl3. The product, 6-chloro-3',5'-di-O-acetyl deoxyguanosine, was converted to O6-methyldeoxyguanosine with sodium methoxide and phosphorylated in the 5' position with carrot phosphotransferase. Monophosphate was converted chemically to the triphosphate and copolymerized with dCTP by terminal deoxynucleotidyl transferase. The resulting template, which contained O6-methylguanine, was tested for its ability to direct RNA synthesis by bacterial RNA polymerase. The presence of O6-methylguanine was shown to lead to the misincorporation of UMP in the product polymer, thus strengthening the hypothesis that O6-methylguanine is a promutagenic base.


Electrostatic complexes of mitomycin C with nucleic acids and polyanions. Reductively activated mitomycin C exhibits strong, non-covalent electrostatic binding to polyanions such as polyvinylsulfate and polyphosphate. The protonated C-2 amino group generated by the reduction is most likely responsible for this type of interaction. At moderate drug and salt concentrations only covalent binding to nucleic acids is observable. This is shown to be guanine-specific in DNA for the first time, as well as in synthetic polyribo- and polydeoxyribonucleotides at 10--20 times higher binding levels than previously tested. At higher mitomycin C concentration, however, strong non-covalent electrostatic binding to nucleic acids also occurs, resulting in a binding ratio up to 1 mol drug bound per mol mononucleotide, although this non-specific binding is relatively inhibited compared to polyvinylsulfate. Salts also have an inhibitory effect on the non-specific binding to nucleic acids. A series of mitomycin derivatives were compared for their binding and cross-linking abilities using DNA as substrate, with the following results: (a) the presence of a basic nitrogen . funtion at C-2 promotes binding, both covalent and electrostatic, presumably by kinetically facilitating the approach between positively charged nitrogen and DNA. (b) The aziridine ring is the major covalent binding site, indispensable for crosslinking and determines the guanine-specificity of the binding.


Excision repair in ataxia telangiectasia, Fanconi's anemia, Cockayne syndrome, and Bloom's syndrome after treatment with ultraviolet radiation and N-acetoxy-2-acetylaminofluorene. Excision repair of damage due to ultraviolet radiation, N-acetoxy-2-acetyl-aminofluorene and a combination of both agents was studied in normal human fibroblasts and various cells from cancer prone patients (ataxia telangiectasia, Fanconi's anemia, Cockayne syndrome and Bloom's syndrome). Three methods giving similar results were used: unscheduled DNA synthesis by radioautography, photolysis of bromodeoxyuridine incorporated into parental DNA during repari, and loss of sites sensitive to an ultraviolet endonuclease. All cell lines were proficient in repair of ultraviolet and acetoxy acetylaminofluorene damage and at saturation doses of both agents repair was additive. We interpret these data as indicating that the rate limiting step in excision repair of ultraviolet and acetoxy acetylaminofluorene is different and that there are different enzyme(s) working on incision of both types of damages.


Incorporation, metabolism and positional distribution of trans-unsaturated fatty acids in developing and mature brain. Comparison of elaidate and oleate administered intracerebrally. Mixtures of [1-14C]elaidic acid and [9,10-3H]oleic acid, as well as [1-14C]elaidic or [1-14C]oleic acid alone, were administered by intracerebral injection to 10-day old and adult rats to examine the relative incorporation and metabolism of trans- and cis-monounsaturated fatty acids in developing and mature brain. Rates and extents of total incorporation of label from trans- and cis-acid into complex lipids were similar. Maximum labeling of the neutral lipid, mainly triacylglycerol and cholesterol, occurred prior to 4 h after injection whereas phospholipid, mainly choline phosphoglyceride, was maximally labeled at 4--8 h after injection. The decrease in labeled phospholipid from 24 to 90 h was greater with elaidate than with oleate. At 8 h labeled elaidic acid was preferentially esterified in the 1-position of all phosphoglycerides whereas labeled oleic acid, in contrast to the positional distribution of endogenous oleate, was randomly distributed. Label from elaidate found in palmitate increased with time to 26% of the total recovered label by 48 h. Thus, although some of the trans-fatty acid is oxidized and its labeled carbon is redistributed, most is incorporated unaltered into complex lipid of brain at rates similar to those for its cis-isomer. The developing central nervous system, therefore, does not metabolically exclude the trans-acid, elaidic acid, from membrane lipids.


Effect of intramolecular fatty acid distribution on aspects of triacylglycerol digestion and absorption studied in vitro. The triacylglycerols from several natural sources have been used as substrates for in vitro assays of three different aspects of fat digestion and absorption. The triacylglycerol mixtures were chosen because they differed widely both in total fatty acid composition and in the intramolecular distribution of fatty acids. The assays were designed to test the rate of hydrolysis with pancreatic lipase, the ability to form insoluble calcium soaps during the hydrolysis, and the ability to form mixed micelles with bile during the hydrolysis. Differences in the rate of hydrolysis appeared to be related to the structure and the triacylglycerols from lard and human milk, both of which have palmitic acid esterified in the sn-2 position, were hydrolysed most rapidly. Similarly the ability to form calcium soaps was related to structure and those triacylglycerols which released saturated fatty acids on hydrolysis favoured calcium soap formation. While there was a range of abilities to form mixed micelles with bile no obvious correlation with structure was detected.


Disialosyl paragloboside. A novel ganglioside isolated from human kidney. A ceramide hexasaccharide was purified from the gangliosides of human kidney. Its structure was studied by methylation analysis, neuraminidase treatment, weak acid hydrolysis and chromium trioxide oxidation. The structure is suggested to be a disialosyl derivative of paragloboside: formula: (see text) Data indicating the possible existence of a corresponding trisialosyl derivative are also presented.


Studies on the thromboxane synthesizing system in human platelet microsomes. The thromboxane synthesizing system from human platelets has been characterized by using arachidonic acid and prostaglandin endoperoxide as indirect and direct substrates. The synthesis of thromboxanes from either substrates catalyzed by the microsomal fraction was monitored by measuring the formation of thromboxane B2 immunoreactivity. Both hemoglobin and phenolic compounds were required for the maximal synthesis of thromboxane B2 from arachidonic acid but not from prostaglandin endoperoxide. Studies on the kinetics of the formation of thromboxane B2 from either substrates indicated that the synthesis of prostaglandin endoperoxide intermediate is the rate-limiting step in the overall production of thromboxanes from arachidonic acid. Effect of the microsomal protein concentrations on the rate of formation of thromboxane B2 showed concave upward relationship suggesting the possible involvement of endogenous stimulator(s) in thromboxane synthetase catalyzed reaction. A variety of compounds including sulfhydryl inhibitors, prostaglandin endoperoxide analog, prostaglandin antagonist, N-0164, and N-substituted imidazoles could directly inhibit the thromboxane synthetase, while nonsteroidal antiinflammatory drugs apparently affected the prostaglandin endoperoxide synthetase which catalyzes the synthesis of the immediate precursor of thromboxanes.


Bioconversion of sitosterol to useful steroidal intermediates by mutants of Mycobacterium fortuitum. A series of mutants which are blocked at various stages of the sterol degradative pathway have been isolated from the potent sterol degrader Mycobacterium fortuitum ATCC-6842. Sitosterol bioconversions by these mutants result in the accumulation of a number of intermediate compounds, some of which are potentially useful as substrates in the manufacture of medically important steroids. These intermediates include androst-4-ene-3,17-dione, androsta-1,4-diene,3,17-dione, ring A-degraded tricyclic compounds and various 9alpha-hydroxy-steroids.


Isolation and preliminary characterization of rat liver cells resistant to 7-ketocholesterol. Two clones of the rat liver cell line, GAI, were selected and propagated in 4 microgram/ml of the cholesterol analogue, 7-ketocholesterol. One of the variants was also found to be resistant to another toxic analogue, 25-hydroxycholesterol. 3-hydroxy-3-methylglutaryl coenzyme A reductase (EC 1.1.1.34) activity was elevated in both variant clones, but was not increased substantially upon incubation of the cells in lipid-depleted medium. The reductase activity was increased 3-fold in the wild type cells under the same conditions. Incubation of the cells in lipid-depleted medium resulted in a 3-fold increase in the rate of sterol synthesis from [14C]acetate in all three types. In the presence of 2 microgram/ml 7-ketocholesterol, the reductase activity in all three cell lines decreased at an equal rate, but activity of the enzyme was still measurable after 48 h in the presence of the analogue. Sterol synthesis was reduced almost to zero in the wild type cells under these conditions, while the variant cells retained 12--15% of their capacity for sterol synthesis. These results are interpreted to indicate that 7-ketocholesterol, besides affecting sterol synthesis at the level to 3-hydroxy-3-methylglutaryl coenzyme A reductase, must also affect sterol synthesis at another step further along the synthetic pathway.