[Change in the content of T-and B-lymphocytes in experimental benzene poisoning (author's transl)]. Changes in T-and B-lymphocytes content in experimental benzene intotication (BI) of CBA-mice were studied. The content of spontaneously rosette-forming cells, B-lymphocytes with C3 -receptors and T-cells in macro-and microtest was determined. It was shown that BI resulted in disorders in the function of thymus, depression of T-cell system and autoimmune shifts. A number of characteristics in kinetics of various fractions of lymphoid cells due perhaps, to different sensitivity of particular types of lymphoid cells to benzene and compensatory response as well. The significance of described phenomena is discussed.


[Allergological rapidtest by measurement of histon methylation in lymphocyte culture (author's transl)]. Measuring an increased methylation of histons in sensitized lymphocytes is a method of detecting allergy against several allergens. Because of the short culture time it is adapted as an allergological rapidtest without high risk for microbial contamination.


Mucolipidosis type III. Multiple elevated serum and urine enzyme activities. The clinical characteristics of a 16-year-old white girl with mucolipidosis type III included early growth retardation, severe dysostosis multiplex, restricted joint motion, tight indurated skin, swollen eyelids, late-onset hepatosplenomegaly, umbilical hernia, corneal opacities, and only slightly impaired mental and neurological development. Cultured fibroblasts contained numerous coarse perinuclear retractile inclusions. Biochemical findings indicated the following: (1) normal levels of urinary acid mucopolysaccharides, (2) deficient activities of multiple lysosomal hydrolases in cultured fibroblasts, (3) elevated activity levels of seven serum lysosomal hydrolases, and (4) elevated activity levels of four lysosomal hydrolases in urine.


Vancomycin therapy of bacterial endocarditis. Fifteen patients with bacterial endocarditis were treated with vancomycin between 1967 and 1976. The indications for vancomycin therapy were penicillin-cephalosporin allergy in six patients, antibiotic resistant bacteria in six, initial therapy in one and culture-negative endocarditis in two. The causative microorganisms were Staph. epidermidis (four patients), Staph. aureus (two patients), diphtheroids (four patients), viridans streptococci (two patients) and enterococci (one patient). Minimum inhibitory concentrations of vancomycin for these organisms ranged from 0.8 to 3.1 micrograms/ml. The patients received vancomycin for two to 10 weeks (mean five weeks). Cure was achieved in 13 patients, including six with prosthetic valve endocarditis (PVE). Two patients had a relapse of PVE and cultures of blood or heart valve were positive within two months of vancomycin therapy. Vancomycin serum levels did not exceed 50 micrograms/ml, and no serious drug toxicity was encountered in any patient. Three patients had minimal audiogram changes beyond the social hearing range. One patient had mild phlebitis and a rash, and one patient had a transient leukopenia. Vancomycin is an effective nontoxic antibiotic in patients with endocarditis when penicillin or cephalosporin therapy is not appropriate.


Childhood impairments and subsequent social adjustment. The relationship of a variety of childhood impairments to subsequent social adjustment measures of occupations, agency utilization, social participation, and arrest records was examined. Generally, results indicated that impaired persons adjust quite well in some respects but not so well in others. There were differential social adjustments by type of impairment and by age, sex, ethnicity, and socioeconomic status. The data analysis suggests complex relationships between impairment and social adjustment.


Speaking fundamental frequency characteristics of institutionalized adults with Down's syndrome. Mean speaking fundamental frequency was obtained from 16 institutionalized adults with Down's syndrome (8 males, 8 females) and from 16 nonretarded adults of similar age and sex. Both male and female adults with Down's syndrome exhibited a higher mean speaking fundamental frequency than did the nonretarded adults of the same sex. This finding does not support early reports of a characteristically low-pitched voice among Down's syndrome individuals.


Patterns of human fetal breathing activity at 34 to 35 weeks' gestational age. Continuous measurements of human fetal breathing movements at 34 to 35 weeks' gestational age were made with an ultrasonic real-time scanner for periods of 24 hours in 11 women. A significant increase occurred in fetal breathing activity during the second and third hour following meals and this pattern apparently followed an increase in maternal plasma glucose concentration. There was a prolonged significant increase in fetal breathing activity between 0100 and 0700 hours. Increases in human fetal breathing activity accompanied by increased gross fetal body movements occurred for periods of 20 to 60 minutes out of every 1.0 to 1.5 hours of observation time. It was concluded that the percentage of time spent breathing by normal fetuses was related to time of day and maternal meals. The alternating changes of fetal activity and inactivity observed may represent biologic changes of sleep state in the human fetus in utero at 34 to 35 weeks' gestational age. It will be important to account for these three patterns of fetal breathing activity in clinical studies which examine the usefulness of fetal breathing movements in assessing fetal health.


Stigmatization of carrier status: social implications of heterozygote genetic screening programs. Possible latent psychological and social consequences ensuing from genetic screening programs need to be investigated during the planning phase of national genetic screening programs. The relatively few studies which have been performed to determine psychological, social, and economic consequences resulting from a genetic screening program are reviewed. Stigmatization of carrier-status, having major psychosocial implications in heterozygote genetic screening programs, is discussed and related to Erving Goffman's work in the area of stigmatization. Questions are raised regarding the relationship between such variables as religiosity and sex of the individual and acceptance of the status of newly identified carrier of a mutant gene. Severity of the deleterious gene and visibility of the carrier status are two important factors to consider in an estimation of potential stigma. Specific implications are discussed for four genetic diseases: Tay-Sachs, Sickle-Cell Anemia, Huntington's disease and Hemophilia.


Prolonged curarisation in the presence of renal impairment. Prolonged curarisation in the presence of renal failure occurred in six cases where pancuronium was used, and one case where alcuronium was used. The cases are presented with a brief review of the literature. Pancuronium must be used with great caution if postoperative reversal problems are to be avoided. Greater use of adjuvants will reduce requirements and may eliminate the problems encountered in renal failure.


Calusterone.

Calusterone. Androgens are effective therapeutic agents in postmenopausal women with metastatic breast cancer. Early studies with testosterone propionate showed objective regression rates of slightly more than 20%. Subsequent work with synthetic androgens has failed to show a significant reduction in toxicity or an increase in therapeutic efficacy over testosterone propionate. Calusterone (7 beta,17 alpha-dimethyltestosterone), in early clinical trials, showed both of these qualities and was believed by some to be the "ideal androgen." As with many new drugs, subsequent work failed to confirm these early findings, and most of the later data gathered on calusterone are inconsistent. We conclude from our knowledge at present that calusterone offers no real advantages or disadvantages over other androgens in the treatment of breast cancer.


Legal ramifications of the medical definition of back disease. Back pain occurring without external force is extraordinarily prevalent in all segments of society. The practicing internist is often faced with the complaint, a need to define its cause, and a request to render an opinion as to whether the patient is disabled in the context of his employment. This opinion will have some weight in determining compensability under workmen's compensation law. This essay examines the inferences, as regards back pain, upon which compensation law is based. These inferences are then tested in light of our current understanding of the pathophysiology of back pain. A number of fallacies become apparent that may provide impetus for remedial legislation and clearly mandate further clinical investigation.


Histochemical changes of the rat testis and epididymis after treatment of alpha-chlorohydrin-effects of a single low dose. A histochemical study has been made of the localization and changes of lipids, carbohydrates, ATPase and 5'-nucleotidase in fresh and fixed frozen sections of testicular and epididymal components in the normal and alpha-chlorohydrin-treated rats. After treatment with a single low dose of alpha-chlorohydrin, the phospholipids are decreased with corresponding increase in triglycerides in both the testis and epididymis. Glycogen, ATPase and 5'-nucleotidase are also decreased after treatment with alpha-chlorohydrin. The physiological significance of these histochemical changes has been discussed.


Thymic lymphocytes in myasthenia gravis. Lymphocytes from the thymus glands of myasthenic patients and controls were characterized in terms of their percentages of B- and T-cells and their ability to stimulate the proliferation of autologous peripheral blood lymphocytes. Percentages of B- and T-cells were the same in the thymuses of myasthenic and control subjects. Autologous peripheral lymphocyte stimulation by thymocytes was observed in only 1 of 3 myasthenic patients, and this was with a thymus gland containing a thymoma. These results thus differ from those of other investigators.


Huntington's disease: clinical and chemical effects of choline administration. Choline (Ch) administration sequentially increases blood Ch, brain Ch, and brain acetylcholine (ACh) levels in the rat and might be useful in Huntington's disease, a brain disorder thought to be associated with deficient cholinergic tone. Ten patients with Huntington's disease took Ch orally (8 to 20 gm per day) after the extent of their disabilities had been documented by clinical examination, movies, and combined electromyogram-accelerometer recordings. Choline levels in blood and cerebrospinal fluid increased markedly during treatment with Ch, affirming that oral Ch administration increases the amount of ACh precursor delivered to the brain. Although some of the patients exhibited transient improvement in speech, balance, and gait, treatment with Ch failed to bring about consistent or lasting improvement in any of the subjects.


Huntington's disease--growth of fibroblast cultures in lipid-deficient medium: a preliminary report. When skin fibroblast cultures derived from subjects with Huntington's disease (HD) were grown in medium supplemented with lipid-deficient serum (LDS), growth rates were less than those of control cultures. The mean percentage increase in generation times for nine control cultures incurred by using LDS rather than fetal calf serum (FCS) was 8.0%. Four cultures from 2 patients with HD failed to grow in LDS but grew well in medium supplemented with FCS. In nine other cultures derived from 7 patients with HD the mean percentage increase in generation time incurred by using LDS was 88.1%. Returning cultures to FCS or adding a mixture of linoleic and linolenic acids restored the growth rate of HD cultures to normal.


Abnormalities in growth of skin fibroblasts of patients with Huntington's disease. Culture characteristics in a variety of media were determined in 18 skin fibroblast cultures from 8 subjects with Huntington's disease (HD) and 20 cultures from 10 control subjects. Skin fibroblasts from HD subjects consistently and reproducibly demonstrated maximal cell densities that were higher than control values. There was no significant difference in growth rates. The relationship of this phenomenon to the HD gene is unknown, as are its specificity and time of appearance in the course of the illness.


Comparison of ribosomal subunit proteins from normal human and Huntington's disease skin fibroblasts. The ribosomal subunit proteins from skin fibroblasts of normal persons and patients with Huntington's disease (HD) were isolated by KCl-puromycin and by EDTA treatments and were compared by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The data indicate that the ribosomal proteins are identical in control and HD fibroblasts.


Inhibitors of GABA metabolism: implications for Huntington's disease. Stereotaxic injection of kainic acid into rat striatum produces neuronal degeneration and neurochemical alterations resembling Huntington's disease (HD). Since correction of the deficiency in gamma-aminobutyric acid (GABA) in HD may be of therapeutic value, the efficacy of drugs that inhibit the GABA-degrading enzyme (GABA-T) in elevating the levels of GABA has been examined in the animal model for HD. High doses of di-N-propylacetic acid only partially correct the GABA deficiency in the striatum, whereas low doses of gamma-acetylenic GABA, a specific irreversible inhibitor of GABA-T, completely restore the levels of GABA in striatum and substantia nigra.


Programmed feeding as a model of chronic alcoholism in the rat. Programmed-feeding polydipsia results in a reliable model of chronic alcoholism in the rat. High oral ethanol comsumption and a predictable withdrawal reaction associated with audiogenic seizures are produced. The maintenance of high blood ethanol levels for three weeks in 18 male Charles River rats was associated with audiogenic seizures after 6 or 8 hours of withdrawal. These chronic alcoholic rats had enhanced blood clearance of ethanol. The cerebral cortical crude mitochondrial fraction showed a decrease in total and magnesium-dependent adenosine triphosphatase activity in alcoholic and control (water-fed) rats compared with normal rats.


Huntington's disease: treatment with muscimol, a GABA-mimetic drug. Muscimol, a gamma-aminobutyric acid (GABA) analogue that exerts potent and specific agonist effects on GABA receptors, was administered orally to 10 patients with Huntington's disease. In this double-blind study, muscimol treatment did not result in improvement in these patients' motor or cognitive functions. However, muscimol administration did ameliorate chorea in the most severely hyperkinetic patient, and it was associated with the appearance of dystonic features, electroencephalographic changes, and behavioral alterations in some patients. These latter observations support a functional relationship between GABA-ergic activity and the genesis of both systonia and EEG abnormalities in humans. The therapeutic failure of muscimol indicates that the GABA disturbances in Huntington's disease does not alone account for the clinical features of this disorder.


[Makeup and properties of E. coli cells with a varying level of resistance to tetracycline]. Lipopolysaccharide composition of tetracycline sensitive and resistant strains of E. coli was studied comparatively. It was shown that that resistance of E. coli to tetracycline was probably due to the differences in the lipopolysaccharide component composition of the outer membrane. On the basis of the activity comparison of the Mg2+- and Ca2+-activated ATP-ase of the membrane fraction of the tetracycline sensitive and resistance strains of E. coli it was concluded that the resistance development in the strains tested to tetracycline was not associated with the changes in the ATP-ase activity.


Fixed drug eruptions to tetracyclines.

Fixed drug eruptions to tetracyclines. We observed 11 cases of fixed drug eruption to tetracyclines. Two cases were in a mother and her adult son, and two patients experienced fading of their lesions while continuing to take 1 g of the drug per day. Several patients were cross-reactive to different salts.


A singular dermatosis of Mongols.

A singular dermatosis of Mongols. A long-term, erythematous, follicular papular dermatosis affecting the presternal and interscapular skin was found to occur in subjects with Down's syndrome. One hundred seventy-four subjects (110 male and 64 female) were surveyed. The condition was seen in 46% of the males, with the highest incidence in patients who were between 20 and 40 years of age; the condition was rare in the females. Our findings suggest that this condition is much commoner than any other abnormal skin feature described in the literature on Down's syndrome.


Dinitrochlorobenzene therapy for alopecia areata. Ninety patients with alopecia areata were treated with weekly applications to one side of the head of dinitrochlorobenzene (DNCB) dissolved in acetone, the other side of the head serving as control region. In 80 patients (89%) hair regrew either exclusively on the treated side, or considerably faster and denser on this side. The difference was noted, in the majority of cases, within eight weeks. The initial response, however, could not be maintained in all of these patients. Persistent response was observed in 72 patients (80%). Peribulbar round cell infiltrates were found to be more constant and denser on the treated side, suggesting that topically applied DNCB affects the peribulbar infiltrate present in alopecia areata. Possibly, the therapeutic result is due to altered local immunoregulation.


Topical 6-aminonicotinamide plus oral niacinamide therapy for psoriasis. Ninety-nine patients with psoriasis were treated topically with 6-aminonicotinamide (6-AN) in four years. In a double-blind study, 1% 6-AN gel was superior to 0.1% triamcinolone acetonide in 29 comparisons, equal in three, and inferior in one when applied without occlusion for four weeks. In an open study, 1% 6-AN was better than four "potent" steroid creams in 34 comparisons, equal in four, and inferior in one. Substantial improvement or complete clearing of plaques occurred in 85 of 99 patients. Tachyphylaxis occurred in ten, but was not permanent. Mucocutaneous toxicity appeared in 25% but was usually easily controlled. Tinnitus occurred in four, but none had deterioration of audiogram readings. The combination of topical 6-AN and oral niacinamide therapy gives promise of an effective and safe treatment for psoriasis. One of three patients with pityriasis rubra pilaris improved considerably with 6-AN.


Localized hemosiderosis as a sequela of acne. Dark blue-black pigmentation has persisted in areas of previous acne scarring in a young woman. By histologic and electron microscopic examination, the pigment is demonstrated to represent hemosiderin. Much of the hemosiderin is lying free within macrophages, not bound by lysosomal membranes that normally would protect the cell from this toxic material.


The preclotting of porous arterial prostheses. A four-step preclotting method is presented for use with porous filamentous Dacron prostheses in the fully-heparinized patient. The method employs controlled fibrin formation within graft interstices, heparin neutralization of all thrombin remaining in the graft wall, and delay of systemic heparin neutralization until 15--20 minutes after clamp release. The resulting flow surface is impervious, smooth and hypothrombogenic. Experimental data are presented which support the rationale of this four-step preclotting method. Four years of clinical experience with the method are summarized, involving 300 prosthesis limbs used in aortic bifurcation, aortofemoral, femorofemoral, axillary-femoral and femoropopliteal positions in 192 patients. A clinical perspective of preclotting techniques is presented in which the proper use of this new method is suggested.


[Mitral prolapse syndrome. Simple diagnosis of forms with severe arrhythmia]. The mitral valve prolapse syndrome encompasses a wide clinical spectrum with a majority of patients presenting with a benign course while a few present with severe arrhythmias. Nineteen patients with a systolic click and angiographic mitral valve prolapse were studied to determine which of several simple tests identified the subgroup with severe ventricular arrhythmias. The incidence and type of ventricular arrhythmias, documented by a 24 hour recording (Holter), were correlated with: 1) cardiac size assessed by the cardiothoracic ratio; 2) echocardiographic indices: end-diastolic septal and posterior wall thickness and left ventricular end-diastolic dimension; 3) a ten minute rhythm strip (mini-Holter). Only patients with ventricular hypertrophy (5 patients) or dilatation (7 patients) had frequent and severe arrhythmias. In these two subgroups, the cardiothoracic ratio was larger than in patients with a normal echocardiogram (7 patients). The ten minute rhythm strip was abnormal in all those patients with severe ventricular arrhythmias (ventricular tachycardia). Thus, in patients with mitral valve prolapse, the presence of an abnormal cardiothoracic ratio, echocardiogram and ten minute rhythm strip identifies patients with potentially severe ventricular arrhythmias. The mechanism of these arrhythmias is still poorly understood; however, a certain number of them may be related to a dysfunction of the autonomic nervous system.


[Biochemistry of myocardium taken at autopsy. Preliminary report]. The findings after biochemical analysis of heart muscle taken at autopsy are given in this preliminary communication. Human myosin is made up of two heavy sub-units and two light sub-units: it is similar to cardiac myosin found in other mammals, but is different in certain characteristics, particularly immunological ones. Tropomyosin is made up of two different sub-units. The normal human heart contains 1 mg of collagen and 130 microgram of desoxyribonucleic acid (DNA) per 100 mg of fresh tissue. The degree of cardiac hypertrophy correlates with the increase total DNA within the heart, and with the lowering of myofibrillary Ca2+ ATPase, the concentration in the collagen remaining unchanged providing there is no ischaemic heart disease. These techniques may be used to quantify several factors, such as the degree of sclerosis or the nuclear mass in ill-understood conditions such as the primary cardiomyopathies.


Location and sequence of atherosclerotic plaque formation in white Carneau and show racer pigeons: reevaluation and redefinition. Sudan staining of gross specimens with correlated histologic sections was used to localize the accumulation of fat in the aortas of genetically susceptible and resistant strains of pigeons. The birds were maintained on a cholesterol-free grain diet, and studies were done sequentially from ages 1 month to 4 years. Evidence suggests that the accumulation of lipids in musculoelastic cushions located at the origin of small branches is not necessarily a precursor of complicated lesions, but that it occurs concurrently in susceptible and resistant strains. In contrast, in the "lesion area," lipid accumulation is more striking and occurs earlier in the susceptible strain. It precedes proliferation by three to six months. Thus, in this model, two distinct types of fatty streaks can be identified and their biologic features can be defined and related to their propensity for atherogenesis.


Symphyseal and sacroiliac joint pain associated with pubic symphysis instability. Fifty patients presenting with lumbosacral and inguinal pain were examined by routine clinical radiographic and electromyographic evaluations. All were without antecedent history of major pelvic trauma or spinal surgery but demonstrated evidence of pubic symphysis instability. Slip between the pubic rami in excess of 2 mm could be demonstrated in each with alternate leg weight bearing. Asymmetry of hip mobility on the symptomatic side with a reduction in abduction and external rotation was present in 20 patients. An approach to treatment of pain associated with pubic symphyseal and associated sacroiliac joint instability is described, combining both intraarticular steroids, lumbosacral supports and physical therapy modalities.


[Tissue specific effect of lymphocytic chalone isolated from rat spleen]. Chalone-containing alcohol extract from the rat spleen consisting of 8--10 substances of protein nature, according to the data of the disk electrophoresis, possesses a selective property to inhibit the entrance of human transformed lymphocytes into the phase of DNA synthesis. Injection of lymphocytic chalones to the mice allotransplants of a donor's skin grafts, which differ from the recipient by a strong antigen, temporally delays the rejection of the transplanted graft. In mice-recipients, the thymus treated with lymphocytic chalone is characterized by the depletion of the cortical substance.


Huntington's disease. Cerebrospinal fluid GABA levels in at-risk individuals. Gamma-aminobutyric acid (GABA) was measured by the ion-exchange fluorometric method in CSF from 22 individuals at risk for Huntington's disease (HD), six individuals with HD, and five neurologically normal controls. The mean (+/- SD) GABA level in the specimens from patients with HD was 142 +/- 27 pmoles/ml, whereas that of the normal control specimens was 297 +/- 87 pmoles/ml. The mean GABA level of the specimens from the individuals at risk for HD was 209 +/- 79 pmoles/ml; however, nine of these were in the normal range with a mean value of 281 +/- 72 pmoles/ml, while the other 13 were below the normal range with a mean value of 159 +/- 27 pmoles/ml. The data indicate that low GABA levels in CSF are evident prior to the onset of symptoms of HD but a predictive value can only be determined by continued observation of the clinical course of these at-risk individuals.


Effect of an opiate antagonist on movement disorders. Animal experiments suggest that opiate peptides might play a role in extrapyramidal function. This hypothesis was tested by administering the opiate antagonist, naltrexone, in doses sufficient to antagonize exogenous opiates, to patients with parkinsonism and Huntington's disease. No improvement in the clinical features of either disorder was noted.


Some problems of the severely handicapped. Acceptance of the severely handicapped in our community is limited. It may be limited by lack of experience, as 'the chicken and the egg' syndrome applies--do we keep the handicapped away from the public because of adverse reaction, or does the public react adversely because the handicapped are kept away? I work with handicapped children and their families, and therefore the following article is related to children.


[Current concepts on gastroschisis].

[Current concepts on gastroschisis]. Present day concepts about history, etiology, embriology, and management of gastroschisis are reviewed and a case is reported; it was treated in two surgical steps. A silastic net was applied and the intestinal loops were gradually introduced until the abdominal defect closed completely. Parenteral feeding was also given.


Removal of intra-abdominal intrauterine contraceptive devices. Thirteen cases of misplaced intrauterine contraceptive devices removed from the abdominal cavity over a two year period at the Chelsea Hospital for Women, are reviewed. Five of the six plastic devices were removable laparoscopically but all the copper containing devices required a laparotomy for removal because of an omental or peritoneal reaction.


The use of aurovertin to determine the F1 content of submitochondrial particles and the ATPase complex. (1) The concentration of aurovertin-binding sites calculated from fluorimetric titrations of submitochondrial particles is equal to the F1 concentration, calculated from the concentration of F1-binding sites in stripped particles. (2) Direct binding experiments show that the fluorescence enhancement of aurovertin bound to submitochondrial particles and the isolated ATPase complex is less (or absent) at higher concentrations than at lower concentrations. The binding data can be described by 'specific' and 'non-specific' binding. The concentration of the 'specific' sites is twice that derived from fluorimetric titrations. (3) After dissociation of the bound F1 with LiCl, fluorimetric titrations with aurovertin yield linear Scatchard plots. The fluorescence enhancement and KD are equal to those of the beta-subunit-aurovertin complex. The concentration of beta-subunits is double the concentration of F1. (4) It is concluded that both for submitochondrial particles and the isolated ATPase complex the most reliable and simple way to determine the F1 content is to dissociate the F1 with LiCl, spin down the insoluble material and titrate the supernatant (containing free beta-subunit) with aurovertin.


Specificity of nucleotide binding and coupled reactions utilising the mitochondrial ATPase. 1. Tightly bound ATP and ADP, found on the isolated mitochondrial ATPase, exchange only slowly at pH 8, but the exchange is increased as the pH is reduced. At pH 5.5, more than 60% of the bound nucleotide exchanges within 2.5 min. 2. Preincubation of the isolated ATPase with ADP leads to about 50% inhibition of ATP hydrolysis when the enzyme is subsequently assayed in the absence of free ADP. This effect, which is reversed by preincubation with ATP, is absent on the membrane-bound ATPase. This inhibition seems to involve the replacement of tightly bound ATP by ADP. 3. Using these two findings, the binding specificity of the tight nucleotide binding sites was determined. iso-Guanosine, 2'-deoxyadenosine and formycin nucleotides displaced ATP from the tight binding sites, while all other nucleotides tested did not. The specificities of the tight sites of the isolated and membrane-bound ATPase were similar, and higher than that of the hydrolytic site. 4. The nucleotide specificities of 'coupled processes' nucleoside triphosphate-driven reversal of electron transfer, nucleoside triphosphate-32Pi exchange and phosphorylation were higher than that of the hydrolytic site of the ATPase and similar to that of the tight nucleotide binding sites.


Activation of partial reactions of the Ca2+-ATPase from human red cells by Mg2+ and ATP. (1) At ATP concentrations up to 30 micrometer addition of 0.5 mM MgCl2 in the reaction mixture increases both the rate of formation and the steady-state level of the phosphoenzyme of the Ca2+-ATPase from human red cell membranes. Under these conditions Mg2+ has no effect on the rate of dephosphorylation, which remains slow. (2) In the presence of Mg2+ the rate of dephosphorylation is increased 5 to 10 times by high (1 mM) concentrations of ATP. (3) Provided Mg2+ has reacted with the phosphoenzyme, acceleration of dephosphorylation by ATP takes place in the absence of Mg2+. This suggests that the role of Mg2+ on dephosphorylation is to convert the phosphoenzyme into a form that, after combination with ATP, reacts rapidly with water. (4) The results are consistent with the idea that combination of ATP at a non-catalytic site is needed for rapid dephosphorylation of the Ca2+-ATPase.


Membrane-bound cooperative enzymes. Stokes' radii, Hill plots and membrane fluidity in the regulation of adenosinetriphosphatase from Escherichia coli. The soluble Ca2+-ATPase from Escherichia coli had a distinctive behavior with respect to inhibition by Na+ measured at 36 degrees C and 19 degrees C. At the first temperature the Hill plots are linear and show a slope of 1.1 while at 19 degrees C the plots are biphasic, with slopes of 1.8 and 0.8 before and after the break, respectively. The break occurs at about 50 nM NaCl. Gel chromatography was performed in jacketed Sepharose 4B columns kept at 2 temperatures in the presence of different concentrations of NaCl. It was found that the Stokes' radius of the enzyme was dependent on the temperature and on the salt concentration. Equilibrium sucrose gradients run at 19 degrees C showed that the sedimentation constant of the enzyme remained constant irrespective of the NaCl concentration used. It is concluded that a "folding" of the enzyme takes place in the presence of NaCl, the process being complete at about 50 mM NaCl at 19 degrees C and at about 20 mM at 36 degrees C. The results are in excellent agreement with the kinetic data: the "folded" or "compact" configuration would show no cooperative response towards Na+ while the "expanded" conformer would present strong cooperativity. This is also in agreement with the results obtained with the enzyme embedded in the membrane: when the membrane is fluid a high n value (Hill coefficient) is found; when the membrane is more rigid the value of n falls. A model explaining all our results is proposed and discussed.


Erythrocyte actin and spectrin. Interactions with muscle contractile and regulatory proteins. Actin and spectrin were isolated from washed red blood cell membranes. Spectrin bound and polymerized erythrocyte actin in the absence of potassium. Spectrin coated into polystyrene latex particles bound 8--9 mol of erythrocyte actin per mol of spectrin when actin was in its depolymerized state. Spectrin enhanced the interaction of erythrocyte actin with muscle myosin as manifested by changes in Mg2+-ATPase activity. A similar enhancement also was observed with muscle alpha-actinin while muscle tropomyosin abolished these effects. The data suggest that spectrin may play the role of polymerizing factor as well as the anchoring site for erythrocyte actin just as alpha-actinin is the anchoring site for actin filaments in muscle and other non-muscle cells.


Studies on the heterogeneity of sarcoplasmic reticulum vesicles. Isolated sarcoplasmic reticulum vesicles from rabbit white muscle were separated into a light (15--20% of total microsomes) and a heavy (80--85%) fraction by density gradient centifugation. The ultrastructure, chemical composition, enzymic activities and localization of membrane components in the vesicles of both fractions were investigated. From the following results it was concluded that both fractions are derived from the membranes of the sarcoplasmic reticulum system of the muscle: (i) The protein pattern of both fractions is essentially the same, except for different ratios of acidic, Ca2+-binding proteins. (ii) The 105000 dalton protein of the light fraction cross-reacts immunologically with the Ca2+-dependent ATPase of the heavy fraction. (iii) Ca2+-dependent ATPase, although of different specific activity, is found in both fractions. After rendering the vesicles leaky, specific activities in both fractions reach the same value. The light fraction was found to consist of "inside-out" vesicles by the following criteria: (i) No Ca2+ accumulation can be measured and the Ca2+-dependent ATPase activity is low and variable. (ii) The rate of trypsin digestion is lower and, compared to the heavy microsomes, a different ratio of degradation products is obtained. (iii) The sarcoplasmic reticulum membrane has a highly asymmetrical lipid distribution. This distribution of aminophospholipids is opposite to that in vesicles of heavy fraction. The light sarcoplasmic reticulum fraction has a higher phospholipid to protein ratio than the heavy one. This is consistent with the possibility that the two fractions derive from different parts of the sarcoplasmic reticulum system.


Properties of three distinct pyrimide transport systems in yeast. Evidence for distinct energy coupling. In Saccharomyces cerevisiae the uptake of cytosine, uracil and uridine is mediated by three permeases. Using mutants blocked in the metabolic utilization of these three compounds we were able to study their specific uptake. Cytosine and uridine show simple saturation kinetics, whereas uracil uptake is a biphasic process. A comparison of the effects of several inhibitors of energy metabolism on these uptake systems was made. Striking differences were found. 2,4-Dinitrophenol (10(-3) M) and NaN3 (10(-2) M) inhibit the entry of the three compounds to similar extent, but chlorhexidine (10(-5) M) and Dio 9 (50 microgram/ml) which are ATPase inhibitors in vitro strongly impaired cytosine and uridine entry and remained without effect on uracil uptake. We provisionally conclude that these systems may be energized by different mechanisms. In the case of cytosine and uridine permease, a membrane ATPase is possibly involved in the process of energetic coupling whereas this does not seem to be so for uracil.


The phospholipid headgroup specificity of an ATP-dependent calcium pump. We have replaced the lipid associated with a purified calcium transport protein with a series of defined synthetic dioleoyl phospholipids in order to determine the effect of phospholipid headgroup structure on the ATPase activity of the protein. At 37 degrees C the zwitterionic phospholipids (dioleoyl phosphatidylcholine and dioleoyl phosphatidylethanolamine) support the highest activity, while a phospholipid with two negative charges (dioleoyl phosphatidic acid) supports an activity which is at least twenty times lower. Dioleoyl phospholipids with a single net negative charge support at intermediate ATPase activity which is not affected by the precise chemical structure of the phospholipid headgroup. The protocol used to determine the phospholipid headgroup specificity of calcium transport protein is novel because it establishes the composition of the lipid in contact with the protein without the need to isolate defined lipid-protein complexes. This allows the lipid specificity to be determined using only very small quantities of test lipids. We also determined the ability of the same phospholipids to support calcium accumulation in reconstituted membranes. Two requirements had to be met. The phospholipid had to support the ATPase activity of the pump protein and it had to form sealed vesicles as determined by electron microscopy. Since a number of phospholipids met those requirements it is clear that in vitro the lipid specificity of the calcium-accumulating system is rather broad.


Neocarzinostatin induction of DNA repair synthesis in HeLa cells and isolated nuclei. The antitumor antibiotic neocarzinostatin that causes DNA strand breaks in vivo and in vitro is shown to induce DNA repair synthesis in HeLa S3 cells. In the repair assay, the parental DNA was prelabeled with 32P and a density label (bromodeoxyuridine) was introduced into the new synthesized DNA. Quantitation of the repair synthesis as measured by the incorporation of [3H]thymidine into the light parental DNA at varying doses of the drug indicate that there is a significant repair response at low levels of the drug (0.2--0.5 microgram/ml) which cause DNA strand breakage and inhibition of DNA synthesis. In isolated HeLa nuclei neocarzinostatin stimulates the incorporation of dTMP many-fold. This enhancement of dTMP incorporation, which requires the presence of a sulfhydryl agent, is a consequence of the drug-induced DNA strand breakage and is in the parental DNA. These results suggest that an intact cell membrane is not required for DNA strand breakage and its subsequent repair.


Stimulation of adenosine triphosphatase activity of sarcoplasmic reticulum by adenylyl methylene diphosphate. The effects of adenylyl methylene diphosphate (AMD), a non-hydrolyzable ATP analogue, were examined in sarcoplasmic reticulum vesicles isolated from rabbit skeletal muscle. The Ca2+-dependent APTase activity measured at 5 degrees C and pH 7.0 in 5.2 micrometer [gamma-32P]ATP and in the absence of added alkali metal salts was stimulated by added AMD. The steady state level of phosphoenzyme, however, was not decreased greatly by added AMP under these conditions. The hydrolysis of the phosphoenzyme formed at the steady state in the absence of added alkali metal salts was accelerated by added AMD to an extent that can account for the stimulation of the ATPase activity. At 5 degrees C and pH 7.0 the maximum stimulation of phosphoenzyme hydrolysis by AMD and the Km value for this ATP analogue were 4.3-fold and 40 micrometer, respectively. These results provide further support for our previous conclusion (Shigekawa, M., Dougherty, J.P. and Katz, A.M. (1978) J.Biol. Chem. 253, 1442--1450) that 2 classes of ATP site exist in the calcium pump ATPase in the absence of added alkali metal salts, one being the catalytic site and the other being the regulation site which activates the activity of the catalytic site.


Interaction of D-fructose and fructose 1-phosphate with yeast phosphofructokinase and its influence on glycolytic oscillations. Fermentation of D-fructose- and D-glucose induced glycolytic oscillations of different period lengths in Saccharomyces carlsbergensis. Recent studies suggested, that D-fructose or one of its metabolites interacted with phosphofructokinase (ATP:D-fructo-6-phosphate 1-phosphofructokinase, EC 2.7.1.11), the core of the glycolytic 'oscillator'. In order to explore the kinetics of interaction, the influence of D-fructose and fructose 1-phosphate on purified yeast phosphofructokinase was studied. D-fructose concentrations up to 0.3 mM stimulated the enzyme, while a further increase led to competitive inhibition. The Hill coefficient for fructose 6-phosphate decreased from 2.8 to 1.0. Fructose 1-phosphate acted in a similar way, up to 1 mM activation and inhibition competitive to fructose 6-phosphate at higher concentration (2.0--3.5 mM) with the same effect on the Hill coefficient. The inhibition patterns obtained with D-fructose or fructose 1-phosphate suggest a sequential random reaction mechanism of yeast phosphofructokinase with fructose 6-phosphate and MgATP2-. The mode of interaction of phosphofructokinase with D-fructose and fructose 1-phosphate is discussed. The influence of both effectors resulted in altered enzyme kinetics, which may cause the different period lengths of glycolytic oscillations.


Frictional properties and molecular weight of native and synthetic myosin filaments from vertebrate skeletal muscle. 1. The molecular weights of a series of synthetic myosin filaments have been measured, using the transport-concentration dependence theory of Rowe, A.J. [Biopolymers, 1977, 16, 2595--2611]. It is shown that for preparations of narrow length distribution (0.60--0.77 micrometer), N, the number of myosin molecules/14.3 nm varies between 3 and 6. 2. The reduced specific viscosity of synthetic myosin filaments has been measured as a function of both concentration and shear rate. From the concentration dependence at zero rate of shear, a value for the "swelling" of the filaments Vs/-v = 2.3 has been calculated. 3. The frictional coefficient of synthetic myosin filaments has been shown to be anomalously but reproducibly high, as compared to that of prolate ellipsoids of the same length and mass. This additional frictional drag has been numerically characterised by a "frictional increment", fi = 1.76 +/- 0.11. 4. A procedure has been devised whereby for any elongated structure which can be assumed to show the same (or other known) fi value, the molecular weight can be estimated from s0 (extrapolated sedimentation coefficient) and 2b (length) alone. 5. An s0 value for natural A-filaments, isolated from rabbit psoas muscle, has been determined by the active enzyme centrifugation technique. From this value, s0 = 132 +/- 3 S, a molecular weight of 1.20 . 10(8) has been computed by the new procedure, for preparations of average length 1.27  micrometer. 6. Contingent upon the validity of the assumptions used (see 4 above) the N value is computed as 3.1 +/- 0.2, consistent with the native, fully intact A-filament having three-fold symmetry, containing 294 myosin molecules, and having a molecular weight based upon myosin and C-protein of 1.31 . 10(8).


An inhibitor from placenta specifically binds urokinase and inhibits plasminogen activator released from ovarian carcinoma in tissue culture. An inhibitor present in placenta and released in placental tissue culture forms specific complexes with each of two molecular forms of urokinase. Autoradiography demonstrated that the inhibitor shifted the electrophoretic position of 125I-labelled urokinase. It did not change the migration of diisopropyl-fluorophosphate-inactivated 125I-labelled urokinase, thereby indicating complex formation dependent on active serine site in urokinase. The inhibitor had a strong neutralizing effect on the plasminogen activators released from human ovarian carcinoma in tissue culture. The placental inhibitor might prove useful in inhibiting the fibrinolytic process necessary for proliferation of tumour vessels.


Pathway of starch breakdown in photosynthetic tissues of Pisum sativum. 1. The aim of this work was to discover the pathway of starch breakdown in the photosynthetic tissues of Pisum sativum. 2. Measurements of the starch in the leaves of plants grown in photoperiods of 12 or 18 h showed that starch, synthesized in the light, was rapidly metabolized in the dark at rates of 0.04--0.06 mumol glucose/min per g fresh weight. 3. The maximum catalytic activities of alpha-amylase, beta-amylase, hexokinase, alpha-glucan phosphorylase and phosphoglucomutase in extracts of leaves showed no diurnal variation in either photoperiod, and exceeded estimates of the rate of net starch breakdown in the dark. 4. Studies with intact chloroplasts, isolated from young shoots and from leaves, indicated that pea chloroplasts do not contain significant activities of alpha-amylase, beta-amylase and hexokinase, although some of the latter may be attached to the outside of the chloroplast envelope. These studies also showed that pea chloroplasts contained sufficient alpha-glucan phosphorylase and phosphoglucomutase to mediate the observed rates of starch breakdown. 5. It is proposed that starch breakdown in pea chloroplasts is phosphorolytic.


Modulation of adenylate cyclase activity by sulfated glycosaminoglycans. II. Effects of mucopolysaccharides and dextran sulfate on the activity of adenylate cyclase derived from various tissues. Heparin was found to be the most potent inhibitor of rat ovarian luteinizing hormone-sensitive adenylate cyclase (I50 = 2 microgram/ml) when compared to other naturally occurring glycosamin oglycans. This inhibition was also apparent when this enzyme was stimulated by follicle-stimulating hormone or prostaglandin E2. Heparin was also found to inhibit glucagon-sensitive rat hepatic adenylate cyclase, and the prostaglandin E1-sensitive enzyme from rat ileum and human platelets. In contrast, heparin stimulated the dopamine sensitive adenylate cyclase from rat caudate nucleus. The sulfated polysugar dextran sulfate exerts similar effects on adenylate cyclase activity of the rat ovary and was shown to inhibit hormone binding to rat ovarian plasma membrane in a manner similar to that exerted by heparin. In contrast to heparin, dextran sulfate inhibited dopamine-sensitive adenylate cyclase from rat caudate nucleus.


[Possible nature of the biogenic amine in the dumbbell-shaped cells of frog taste buds]. An attempt was made to identify specific monoamines contained in the dumb-bell shape cells of the frog taste bud by means of histochemical analysis. It was shown by fluorescent microscopy that preliminary administration of exogenous serotonin into the blood channel of frog tongue resulted in a sharp increase of specific fluorescence of the dumb-bell shape cells, whereas serotonin synthesis inhibition with p-chlorphenylalanine led to reduction and elimination of specific fluorescence. It was concluded that-specific monoamine of the dumb-bell shape cells was possibly of serotonin-like nature.


Electrocardiography: right ventricular hypertrophy. Not only is RVH difficult to diagnose electrocardiographically but it is frequently mimicked by true posterior infarction. The R/S-amplitude ratio in lead V1 and the S wave in lead V5 are the most useful indices of RVH. The ECG in chronic lung disease can be distinguished from that in RVH and is more often confused with that of anterior infarction. The ECG changes found in pulmonary embolism are the same as those found in RVH but they are transient; they are more easily interpreted by means of serial ECGs. In conjunction with clinical data the ECG can be used to predict the severity of pulmonary valve stenosis.


Controlled trial of mobilisation and manipulation for patients with low back pain in general practice. Ninety-four patients with non-specific lumbar pain who were seen by their general practitioners took part in a double-blind controlled trial to compare mobilisation and manipulation with placebo physiotherapy. Immediately after treatment most patients showed improvements in the various features studied, but for several features improvement was more common in the treated group than the controls. At three months the condition of most patients was still improved but the differences between the two groups had largely disappeared. At one year the groups were identical. Prognostic presenting features were sought, but only a shorter history correlated with clinical improvement. This study indicates the high rate of spontaneous resolution of low back pain. A course of mobilisation and manipulation may hasten improvement but does not affect the long-term prognosis.


A critical look at delayed intestinal motility in gastroschisis. The motility of the gastrointestinal tract after the accepted management of gastroschisis by using a Silon pouch and gradually reducing the herniated intestine is compared with the intestinal motility after primary reduction and closure of the defect. Since 1970, 46 children with gastroschisis have been treated at the Hospital for Sick Children, Toronto. Twenty-seven were treated with a Silon pouch. Eleven died after an average postoperative survival of 17 weeks; not one had bowel motility that allowed oral feeding. Among the 16 survivors an average of 10 weeks elapsed between operation and the institution of full oral feeding. Seventeen children were treated by forceful stretching of the abdominal cavity, primary replacement of the eviscerated intestines and closure of the defect. Two died in the early postoperative period of conditions unrelated to the gastroschisis or to the procedure. The average period between operation and full oral intake of food was only 4 weeks. In addition, two neonates were admitted after primary closure was done at the referring hospital. Roentgenograms obtained postoperatively in 24 of the patients showed the "typical" dilated loops with air fluid levels in patients treated with a Silon pouch. The authors' findings indicate that primary reduction and closure of the defect has a distinct advantage over use of a Silon pouch in the treatment of gastroschisis.


Purification of immunosuppressive factors extracted from bovine spleen (lymphoid chalone). A low molecular weight immunosuppressive factor FA which is able to reduce the blastic transformation capacity of lymphoid cells from treated mice has been characterized. It was prepared from a bovine spleen acetone powder and found to be associated partly with high molecular weight carriers in the form of an active complex characterized previously as part of a 'lymphoid chalone' fraction. FA may be obtained by selective ultrafiltration of F followed by P-2 Biogel chromatography of the ultrafiltrate. Thymidine, deoxyinosine and deoxycytidine have been identified as the major constituents of FA by mass spectrometry, ultraviolet absorption data and thin layer chromatography. However, none of these nucleotides has the biological activity of FA.


Purification of immunosuppressive factors extracted from bovine spleen (lymphoid chalone). II. Biological activity. A highly purified extract from bovine spleen (FA) possessing immunosuppressive capacity was studied. Its tissue specificity was analysed in vivo and in vitro and the main components of FA, the well-known nucleosides deoxycytidine, deoxyinosine and thymidine, were tested in vivo. It seems that the molecule responsible for the biological activity of FA is a new substance endowed with strong specific activity and that lymphoid tissue is highly sensitive to the inhibiting capacity of this purified fraction.


[Direct electric measurement of the functioning of adenosine triphosphatase of submitochondrial particles of beef heart]. A direct potential measurement of the activity of bound ATPase has been achieved with beef heart submitochondrial particles adhering to a thick phospholipidic membrane. Electric signals (90 mV amplitude) were found to be fully sensitive to ATPase inhibitors and uncouplers.


[Frequencygrams due to the stimulation of dynamic gamma axons exerting type II effects]. Frequencygrams are graphs giving indirect information on the contraction of intrafusal muscle fibres. Frequencygrams of Cat spindle primary endings (flexor hallucis longus, soleus and peroneus brevis muscles) were recorded during repetitive stimulation of dynamic gamma axons exerting type II effects. The features of these frequencygrams (periodic increments whose frequency is equal to that of the stimulation and "driving") show that they can be ascribed to a relatively strong and fast intrafusal contraction. Two interpretations are considered.


[Acute arthritis of knee with intra-articular presence of D.medinensis. Comparative clinical and immunologic evolution. 1 case]. The authors report a case of knee acute arthritis with intra-articular presence of microfilariae D. medinensis observed in a black African worker immigrated in France. It is a classical manifestion of dracunculosis articular complications which are however less frequent than intra-articular reaction arthritis without microfilariae, or than pyogenes arthritis due to neibourhood superinfections. The authors emphasize the significance of a double clinical and immunological supervision in the course of a convenient treatment including antibiotics, anti-inflammatory agents, local treatment, and rehabilitation.


Assay of an antitumor protein, neocarzinostatin, and its antibody by fluorescence polarization. I evaluated use of the fluorescence polarization technique to measure neocarzinostatin, a proteinaceous antitumor antibiotic, and its antibody, in serum. The antigen (neocarzinostatin), labeled with fluorescein isothiocyanate, was allowed to interact with its antibody in a cuvet, in the instrument, yielding an increase in the fluorescence polarization value. Antibody content was determined in the presence of a definite amount of the labeled antigen, fluorescence polarization values increasing in parallel with each addition of antibody. Antigen content was determined with a known amount of antibody, which reacted at first with an unknown amount of antigen in samples, followed by addition of a definite amount of the labeled antigen (competition). I used the method to determine a pharmacokinetic parameter, the apparent volume of distribution for neocarzinostatin in rabbits, using drug-injected rabbit sera. I evaluated precision, accuracy, and reproducibility, using various samples or possible interfering substances such as bilirubin and hemoglobin, and also compared results for antigen with those by single radial immunodiffusion assay. The present assay is fast (less than 2 min), sensitive (less than 10 nmol/liter can be detected), and simple (there is no separation step before readout of the results).


An electronic aid for teaching writing and drawing skills. Conventional methods for the teaching of writing and drawing skills among the handicapped are highly teacher-intensive. This paper describes an electronic drawing aid which, after an initial teaching session, may be used by an unsupervised patient to develop his drawing and writing capabilities. A major feature of the system is its flexibility in that its performance may be tailored to the needs of any particular patient, in terms of both the degree of complexity of a required drawing and in the level of ability required by the patient to reach a desired level of attainment.


Endocrine effects of spironolactone in man. A double blind, controlled study was carried out in order to investigate the effects of administering spironolactone, 200 mg daily, to five healthy male volunteers. The patterns of change in plasma testosterone (T) and luteinizing hormone (LH) after spironolactone were significantly different from placebo and there were significant increases in the urinary excretion of androsterone (A), aetiocholanolone (EC) and total oestrogen. Urinary dehydroepiandrosterone (DHA) excretion, after an initial rise, declined progressively during the treatment period relative to controls. The results are discussed in the light of previous observations. It is concluded that treatment with spironolactone for 2--4 days will lead to a transient rise in plasma T and urinary DHA. Continued treatment (4--10 days) is thought to cause increased LH secretion, with normalization of plasma T and DHA excretion. These changes are accompanied by increased androgen catabolism and a slightly increased conversion of androgens to oestrogens. Healthy men may therefore show alterations in sex steroid metabolism if treated for several days with high doses of spironolactone.


Immunodeficiency in Down's syndrome: T-lymphocyte subset imbalance in trisomic children. In fourteen children with Down's syndrome the percentage of circulating T-cells forming rosettes with sheep erythrocytes, either in the presence or in the absence of foetal calf serum, was significantly lower than in appropriately matched controls. In contrast the percentage of T-cells forming rosettes with human erythrocytes was significantly higher in children with Down's syndrome than in controls. These data support the hypothesis that a defective T-cell maturation is an early integral feature of Down's syndrome.


Effect of retinyl acetate on sulfated glycosaminoglycan biosynthesis in dermal and epidermal cells in vitro. The effects of retinyl acetate on the biosynthesis of sulfated glycosaminoglycans in dermal and epidermal cells isolated from newborn mice was investigated. Three compartments were analysed for [35S]-glycosaminoglycans; the culture medium, the cellular matrix, and the cells. The individual levels of chondroitin 4-sulfate, dermatan sulfate and heparin and/or heparan sulfate in these compartments as a function of retinyl acetate was also analysed. The addition of retinyl acetate resulted in a dose dependent increase 35SO4 incorporation in the cellular and matrix compartments of the dermis in vitro. At the optimum concentration of 1.8 X 10(-6) M, this increase was 50%. The levels of35SO4 incorporated into medium glycosaminoglycans were relatively constant. There were also changes in the levels of the individual sulfated glycosaminoglycans. The glycosaminoglycan profile was modified differently in each of the three compartments analysed. In the epidermal cells, retinyl acetate at an optimum concentration of 1.8 X 10(-6) M resulted in a dose dependent increase in 35SO4 incorporation in the cellular, matrix and medium compartments. There was no apparent change in the glycosaminoglycan profile, with heparin and/or heparan sulfate being the major sulfated glycosaminoglycan.


Technical note: an improved McCutchen miniviscometer for connective tissue fluids. McCutchen's miniviscometer is a Couette-type instrument that requires only 0.2 ml of synovial or other test fluid. The modification described here adds to the capabilities of the original: control of temperature good to +/- 0.2 degrees C; increase of the maximum measureable viscosity at low shear rates (20 sec-1) to 3000 centipoise; and expansion of the usable range of shear rates to (20--1500 sec-1. The device is rugged and data are reproducible to +/- 1.2% (S.D.) of the mean over the full range of shear rates.


Segmental differences in morphogenetic activity of arterial smooth muscle cells. Histochemical and radioautographic studies. Differences in morphogenetic and metabolic activities of the arterial smooth muscle cells (s.m.c.) of the young rat's aorta and femoral artery were studied by histochemical, radiochemical and quantitative radioautographic methods. 3H-proline was found to be incorporated into the medial myocyte of both vessels and released into the extracellular connective tissue matrix during the first 6 hours. The intracellular and extracellular phases of this process were similar to those of other scleroprotein-synthesizing cells. The 3H-proline incorporation, the metachromasia (GAG) and the activities of acetyl-cholinesterase, beta-glucuronidase, aryl-sulfatase and 5'-nucleotidase were more intense in the aortic media. On the other hand, some oxido-reductases linked with cellular respiration, glycogenolysis and energy production as well as the myosin-ATPase and MAO activities are more intense in the femoral artery. These differences suggest the morpho-functional diversity of the arterial s.m.c.: greater morphogenetic activity of the aortic myocyte; earlier and higher contractile differentiation of the femoral one.


Further observations on the pathological responses of rat skeletal muscle to toxins isolated from the venom of the Australian tiger snake, Notechis scutatus scutatus. 1. Some aspects of the response of mammalian skeletal muscle following the injection of purified toxins from the venom of the Australian tiger snake, Notechis scutatus scutatus, are described. 2. The toxins used were notexin, notechis II-5, notechis II-1 and a modified form of notexin (PBP-notexin). They were injected into the dorso-lateral aspect of one himd limb so that the soleus muscle would be exposed to the toxins. 3. Within 1 h after the injection of notexin, the soleus muscles were oedematous and by 3--6 h, polymorphonuclear leucocytes had entered the interstitial spaces. The invasion of necrotic muscle fibers was extensive by this time. Muscle spindles appeared relatively unaffected by the toxin. 4. The muscle regenerated via myoblasts at 2--3 days to myotubes at 3--5 days, immature muscle fibers at 7--14 days and fully differentiated muscle fibers by 21--28 days. Even after 6 months, however, the nuclei of many muscle fibres remained in a central position. 5. A second component of Australian tiger snake venom was also found to be myotoxic. It was slightly less potent than notexin, but caused qualitatively similar damage to that caused by notexin. It was identified as notechis II-5. A third fraction, notechis II-1, was found to be inactive. 6. Notexin could be neutralized by incubation with tiger snake antivenene; the simultaneous injection of antivenene with notexin did not afford complete protection against muscle damage.


The effect of halides (NaCl and NaI) on in vitro pancreatic elastase activity. The effect of I- and C1- on the enzymatic degradation of elastin pancreatic elastase has been studied. At low halide concentrations both ions show a marked stimulation of elastolysis, I- being significantly more effective than Cl-. Kinetic plots of enzyme activity indicate that anions enhance the susceptibility of the substrate to enzymatic and non-enzymatic degradation. If the halide concentration is raised inhibition of elastolysis occurs starting at 30 mM NaCl and 300 mM NaI. This inhibitory effect is explained by a direct attack of the halides leading to increasing destabilization and inactivation of the enzyme.


Special topical approach to the treatment of acne. Suppression of sweating with aluminum chloride in an anhydrous formulation. A new topical approach to acne treatment--the use of aluminum chloride hexahydrate in anhydrous ethanol (ACAE)--was studied in 141 patients. Using sequential treatment schedules, paired comparison techniques, and various concentrations of ACAE, we established maximal efficacy with minimal local irritation for the 6.25% strength solution. Clinical efficacy and lack of toxicity of this formulation were confirmed by the additional clinical study of 65 patients. The antiperspirant and antibacterial actions of 6.25% ACAE solution were then verified on acne skin areas. It is postulated that the clinical improvement in acne that follows the topical use of ACAE results from one or both of these actions.


[Conservative therapy of post-traumatic splenic rupture]. Our experiences with conservative treatment of traumatic rupture of the spleen in children are described. Laparoscopy is an important diagnostic aid, which also influences the indication for the conservative procedure. An overwhelming, mostly lethal sepsis arises in a small percentage of splenectomized patients. In children, therefore, we favor preserving the traumatized spleen, even if ruptured.


A flowmeter for respiratory monitoring.

A flowmeter for respiratory monitoring. A variable-orifice pneumotachograph is described. It has a resistance that is relatively constant over a flow range from 0.1 to 2.5 liter/sec so it puts out a differential pressure signal that is directly proportional to flow. It is specifically designed for monitoring respirator patients and sacrifices some of the accuracy of a laminar flow (Fleisch) type device to gain greatly improved immunity to artefacts caused by airway water.


Present status of the infant of very low birth weight treated in a referral neonatal intensive care unit in 1974. In 1974, 250 infants with birth weights of 1500 g or less were referred to the Intensive Care Unit at the Hospital for Sick Children, Toronto. Sixty-six per cent survived. Prospective follow-up studies of the survivors have been completed over a minimum of two years. Infants at highest risk for handicapping defects (about 85%) are those who survived intracranial haemorrhage during the neonatal period. Neurological defects encountered in these children were severe--hydrocephalus, spastic quadriplegia, hemiplegia or microcephalus--and were usually associated with low psychometric scoring. The second highest risk group were small for gestational age infants. Of these, 53% were significantly handicapped at two years of age, and 43% had developmental quotients of less than 80. These defects could not be related to postnatal events. The third category at high risk were infants with birth weights of 1000 g or less. Among these, 30% had a significant handicap by two years of age. The outcome in the remainder of the sample was generally good. These results suggest that the decreased mortality of the very premature infant is associated with an increased number of relatively normal survivors and that the infants at highest risk for subsequent defects can be identified at or shortly after birth.


Outcome for infants at high risk of major handicap. Perinatal intensive care had been introduced in University College Hospital, London, by 1966. In the succeeding 10 years, 28-day mortality rates fell among infants of birth weight 1500 g or less. Among the survivors, the incidence of major handicap was 10% or less and the mean IQ increased to within the range expected for a normal population. Of the children aged 8 years or more 76% had no handicaps and were attending normal schools; 18% had minor handicaps or problems for which they were receiving extra help in normal schools; and only 6% were attending special schools. Throughout the 10 years of the study, the overall prognosis for these infants of very low birth weight improved significantly. Results among other high-risk groups were equally encouraging. Analysis of variance of the data from the infants who weighed 1500 g or less at birth indicated that perinatal complications, particularly illnesses associated with abnormal neurological signs or acidaemia in the infants around the time of birth, were the principal factors determining the condition of the survivors at follow-up. Thus, it is likely that additional refinements in the management of the perinatal period may result in further improvements in the prognosis of these and other high-risk newborns.


Application of cost-benefit analysis to programmes for the prevention of mental handicap. The paper is a discussion of the role of cost-benefit analysis in evaluating programmes for preventing mental handicap. It is divided into three broad sections. In the first section the technique of cost-benefit analysis is defined and the possible variants considered. It is concluded that policies for the prevention of mental handicap are most appropriately analysed using a mixture of monetary and non-monetary measures of outcome. The second section lays down some guidelines for undertaking a study: the importance of specifying alternatives to the policy being analysed; specifying input-output relationships; counting only the appropriate costs and benefits; and discounting costs and benefits which occur in the future. The third section looks at the current state of the art in the UK. It concludes that little cost-benefit analysis has been done in the field of mental handicap, indicates where work could usefully be directed, and highlights information that would have to be available before a cost-benefit study could give useful results.


Perinatality: French cost-benefit studies and decisions on handicap and prevention. The Perinatal Care Programme (or 'Périnatalité') was implemented between 1970 and 1975 by the French Ministry of Health, with the aim of reducing the number of birth-related accidents, i.e. the number of deaths and injuries occurring during the perinatal period. It aimed to reduce the perinatal mortality rate from 26/10(3) in 1970 to 18/10(3) in 1980. To give an order of priorities to the actions, a cost-effectiveness evaluation was made. The ratio (Cost over 15 years)/(Number of deaths + number of handicap cases avoided during the same period) indicated that first place should go to resuscitation in the labour room, which cost only 200 francs to save a life while avoiding after-effects. The other measures concerned the training of medical personnel, the development of statistics and research, inoculation against German measles, intensification of antenatal supervision, improvement of supervision during confinement, and the creation of intensive resuscitation units. Evaluation of the programme is still going on. The perinatal mortality rate has considerably decreased, with the objective foreseen for 1980 already reached in 1975. The handicaps will be traced through the results of obligatory screenings at birth, nine months and two years.


Severe mental retardation in Swedish children born 1959-1970: epidemiological panorama and causative factors. The findings from two recent Swedish retrospective studies on severely mentally retarded children born 1959-70 are surveyed. One study concerned an average county--Uppsala--in the middle of Sweden, the other a vast northern county, Västerbotten, with a sparse population. Figures for incidence and prevalence are given, as well as the percentage distribution of aetiological/pathogenic groups and subgroups, and the frequency of multihandicaps. Prenatal causes were apparent in about 70% of affected children, Down syndrome being the main condition (32%). A background of mutant genes in severe mental retardation was found significantly more often in Västerbotten than in Uppsala (17% and 7% respectively). Perinatal causes were revealed in 10 and 8% (9% of the two series pooled together) and postnatal causes in only 3 and 1% (2%). Associated CNS handicaps were present in about half of the children (42% in Uppsala and 52% in Västerbotten), with epilepsy in 30 and 36%, cerebral palsy in 18% and 19%, and a combination of epilepsy and cerebral palsy in 8 and 16%, respectively.


Main causes of major mental handicap: prevalence and epidemiology. The prevalence of educational subnormality of a severe form (between 3 and 3.6 per thousand children of school age) and the prevalence of cerebral palsy (between 2 and 2.4 per thousand) have been fairly stable up to recent years. This stability has also applied to the relative proportions of the different major causes contributing to the handicaps. Where the ascertainment of such conditions is good, their prevalence monitored and the life expectancy of affected individuals estimated, any changes in prevalence can be used to measure the effectiveness of new forms of prevention, or alternatively to indicate the existence of new environmental hazards. Only a multi-pronged campaign against many of the recognized causes will have a substantial impact on prevalence.


[Fibronolytic treatment with activator: clinical results, angiographic findings and laboratory results (author's transl)]. Two-day treatment with activator (equimolar streptokinase-human plasminogen complex) was given to 26 patients. Indications were chronic arterial occlusion and stenosis of the lower-limb arteries (25 patients), and thrombosis of the subclavian veins. The onset of claudication had on average been 7.5 months previously, average walking distance 288 metres. Five occlusions of iliac arteries and three of femoral arteries were removed. In addition, stenosis of the aorta (one case), the iliac artery (ten cases) and femoral artery (one case) was partly removed. Occlusion of the axillary vein was also removed. Average duration of the successfully treated occlusions was 4.3 months. Those stenoses that responded to activator had the well-known morphological criteria for fibrinolysis. The walking distance of the patients increased from an average of 288 to 401 metres. Activator treatment was well tolerated. No serious, especially cerebral, complications occurred. The most striking laboratory finding was of circulating plasmin during the entire duration of the infusion.


Premature closure of the ductus arteriosus (P.C.D.A.): a possible cause of intrauterine circulatory failure. In the course of some 800 perinatal necropsies (corresponding to approximately 26,000 deliveries), 3 stillborn infants were found to have an almost completely closed ductus arteriosus. Each of these showed cardiomegaly, dilatation of right-side chambers, pulmonary hyperaemia and oedema of varying degree and extent; intrauterine death was considered to be due to P.C.D.A. In several further cases the ductus was also narrowed but not necessarily responsible for intrauterine death. This frequency suggests that P.C.D.A. is more common than might be surmised from very scanty documentation in the literature. Possible aetiological factors of P.C.D.A. as well as its effects on the circulation are briefly discussed.


Early phosphate burst of heart myosins- Phylogenic variations. The phosphate burst of cardiac myosins from different animal species was studied by a rapid quench-flow technique. The rate constant of the phosphate burst varied from one species to another. Moreover it was lower for cardiac myosins than for skeletal myosin. The phosphate burst rate correlated with the ATPase in the steady state and with the muscle-shortening speed. By contrast, the amplitude of the phosphate burst did not vary significantly. Its value was 0.8--0.9 per myosin site for each of the myosins studied. Deviations from this stoichiometry were related to the mode of preparation of the myosin and to denaturation by a high pH.


Delayed cutaneous hypersensitivity in children with severe multiple handicaps treated with phenytoin. Peripheral lymphocyte counts, T-cell and B-cell populations, DNCB skin reactions, Mantoux reactions, mitogenic transformation using PHA, and determination of serum phenytoin concentration were carried out in 45 children with severe handicaps. Twenty two of these patients were receiving phenytoin treatment. When compared with control subjects, the skin reactions were significantly depressed in the handicapped children, amongst whom those treated with phenytoin had more impaired reactions than those who were untreated. No significant relationship between the results of the skin reactions and the other parameters was found.


Clinicians, the Mount Sinai program and the Veterans' Administration program evaluated against clinico-pathological data derived independently of the electrocardiogram. 221 electrocardiograms (ECGs) recorded from adult subjects with cardiac symptomatology were interpreted by clinicians and the Mount Sinai (MS) and Veterans' Administration (VA) computer programs. After the clinicians had eliminated their interobserver variability, their interpretations and those of the computer programs were compared with the corresponding clinico-pathological data that had been derived independently of the ECG. A measure of overall diagnostic accuracy was used which showed that the performance of the 2 computer programs was similar and somewhat worse than that of the clinicians. The clinicians and the computer programs were not as good at diagnosing the ECGs of women as at diagnosing those of men. As the VA program's interpretations may be altered by including information about the patient's provisional diagnosis, 6 different sets of these prior probabilities were used to analyse 141 of the original 221 ECGs. Different fixed sets of prior probabilities made only a small difference to overall diagnostic accuracy but a marked difference to the VA program's ability to differentiate Normal from Abnormal. Optimizing the prior probabilities for the individual subjects increased the VA program's overall diagnostic accuracy up to that of the clinicians. Both computer programs correctly diagnosed sinus rhythm as the dominant rhythm in 165 out of 177 subjects. The MS program diagnosed the dominant rhythm as Atrial Fibrillation in 35 out of 41 subjects and the VA program in 27 (X(2) = 3.24; not significant). ECG interpretation; Mount Sinai program; Veterans' Administration program; independent clinico-pathological data; sex differences in the ECG; prior probabilities.


The permanent form of junctional reciprocating tachycardia: further elucidation of the underlying mechanism. A case of the permanent form of junctional reciprocating tachycardia studied by endocardial and epicardial mapping is presented. The ability of the retrograde impulse to penetrate the atrium in 2 different areas, one near the region of the recorded His potential and the other near the orifice of the coronary sinus, is demonstrated. Retrograde conduction persisted after the surgical creation of complete antegrade heart block suggesting the existence of an accessory ventriculo-atrial connection. The decremental properties of retrograde conduction observed suggest that an accessory ventriculo-artrial nodal structure may be the underlying substrate of this arrhythmia.


[Reproductive lifespan and reproductive performance in SPF C3H mice: the onset of reproductive life and production efficiency (author's transl)]. To improve the production system, the onset and the termination of reproductive life of C3Hf/HeMsNrs mice mated immediately after weaning and reared for 400 days of life, were studied. From weaning females mated with a full grown male (group A), the first litter was obtained at a mean age of 47 days, suggesting the first copulation at 26 days of age. The age of males at the first copulation was estimated to be at 44 days of age from the age giving the first litters in weanling males mated with weanling (group B) and full grown (group C) females. The sex ratio of litters delivered by young dams tended to be excess in males. The reproductive performance of dams in later life was not affected by the parturition in earlier age. The production efficiency with weaned youngs per pair during the first 200 days after mating was the highest in group A. It was found from these results that the C3H females attained their sexual maturity at 5 to 6 days after weaning, being available for breeding without any deletion in reproductive performance.


Cerebral changes in the course of intoxication with mercury phenylacetate. A histoenzymic study of cerebral phosphatases and esterases was performed on rats subjected to experimental intoxication with mercury phenylacetate. Following intragastric application of mercury phenylacetate to experimental animals, decreased activities of cerebral ATPase, acP and AChE were observed. The intoxicated animals displayed enhanced cerebral TPPase and partially also NsChE activities. Apart from changes in the histoenzymic pattern of the experimental brains, the ingestion of mercury phenylacetate brought about evident morphological changes in form of neuronal vacuolisation and spongious degeneration of the white matter. The extent of morphological as well as histoenzymic alterations was dependent on the duration of the experimental poisoning.


Adenosine triphosphate and diphosphoglycerate levels in red blood cells from patients with Down's syndrome. The levels of ATP and ATP plus DPG were significantly elevated in erythrocytes from Down's syndrome patients when compared to erythrocytes from age matched controls. The hemoglobin content and hematocrit values were significantly reduced. The resultant tendency towards anemia probably explains the elevation in metabolite levels.


Lack of relationship between Langerhans cells, epidermal cell proliferation and epidermal G1 chalone. Mouse tail interscale epidermis contains numerous Langerhans cells, whereas the adjacent scale regions are devoid of these cells. No difference in a) proliferative activity and b) inhibitory effect of the epidermal G1 chalone can be demonstrated in both regions. A direct relationship between Langerhans cells and growth control may be excluded.


Tubal sterilization by a combined laparoscopic and external technique: preliminary report on 75 cases. A preliminary report is given on a new laparoscopic sterilization procedure. By a double-puncture method, a loop of tube is brought out of the abdomen through the suprapubic incision. The loop is then resected by a modified Pomeroy method. Between Octorber 1976 and June 1978, 75 sterilizations were performed without any major problem. Sterilization failed in one case when one tube could not be seen. All recoveries were uneventful. Safety, effectiveness, simplicity, and potential reversibility seem to be the qualities of this method.


[In vitro stimulation of peripheral blood lymphocytes with allogeneic lymphocytes (mixed lymphocyte culture, MLC) or phytomitogens in patients with Hashimoto's thyroiditis (author's transl)]. To investigate the role of peripheral blood lymphocytes in thyroid auto-immunity, the in vitro responsiveness to allogeneic lymphocytes (MLC) or phytomitogens was examined in the lymphocytes from patients with Hashimoto's thyroiditis. The MLC responses of 16 patients were compared with those of age- and sex-matched normal controls. All possible combinations between lymphocytes from the patients and controls were mixed in both one-way and two-way MLC. From the results of one-way MLC, the stimulatory capacity and responding capacity of lymphocytes from the patients and controls were calculated respectively as follows: : formula: (see text) To the same allogeneic stimulation of Nm (mitomycin-treated stimulating lymphocytes from controls), lymphocytes from the patients (H) exhibited a lower response (P greater than 0.05) than did those from the controls (compare (1) with (3)). Phytomitogen-response was simultaneously studied for residual cells of MLC. Lymphocytes from the patients showed a significantly decreased (P less than 0.01) response to phytohemagglutinin-p and concanavalin A  in comparison with the response of normal lymphocytes. From these results, it was suggested that cell-mediated immunity in vivo was impaired in Hishimoto's thyroiditis.


Myometrial cells in primary culture: characterization and hormonal profile. A method for primary culture of ovine myometrial cells is described. After dissection, myometrium of ewe uteri was digested in trypsin and collagenase. The cells were preplated for 1 h at 37 degrees C. The non-attached cells were grown in appropriate medium supplemented with 2% fetal calf serum. They had a doubling time of 3 days, reached confluency at 10 days and did not exhibit contact inhibition. Cultures were maintained up to 22 days. Characterization of the cells was achieved by electron microscopy, analysis of myosin in cell extracts and assessment of hormone sensitivity. The cells were found to contain myofilaments, characteristic of smooth muscle. A high content of myosin (6--13%) was demonstrated on SDS-polyacrylamide gel electrophoresis: this was confirmed by ATPase activity assay. Cells responded to estradiol stimulation by increased protein synthesis, and bound [3H]estradiol in a specific and saturable way. These results suggest that myometrial cells grown in primary culture should provide a useful model for studying the hormonal control of contractile protein synthesis.


Body temperature and 5-hydroxytryptamine during early postnatal maturation in mice. A relationship between the maturational increases in body temperature and brain indoleamines has been explored through altering brain 5-hydroxytryptamine (5-HT) content. Effects on body temperature at approximately constant ambient temperature (about 24 degrees C) have been studied following pharmacological manipulations in mice up to 16 days postpartum. Administration of 5-hydroxytryptophan (5-HTP) to increase 5-HT levels was associated with decreased body temperature throughout the maturational period. Depletion of 5-HT with p-chlorophenylalanine (p-CPA) was associated with increased body temperature at all ages studied, although 5-HT depletion was less effective in young animals than in older animals. The drug NSD-1034 acts at a different enzymatic step, and is effective in both central and peripheral tissues to reduce 5-HT and norepinephrine (NE) levels. NSD-1034 decreased body temperature up to 10 days of age; however, the effect was reversed at about 14 days, and the drug increased body temperature significantly in 16-day-old animals. These results suggest a role for 5-HT in the mechanisms of heat production in early postnatal life.


[Investigation of the fibroblast chalone (author's transl)]. By ultrafiltration of a cytoplasm preparation from diploid human fibroblasts (Flow 2000) we obtained a fraction which inhibited the proliferation of the same cells. We succeeded in demonstrating fibroblast chalone, because the activity was endogenous, reversible and affected murine fibroblasts, but not human cervical carcinoma cells (HeLa S3) or neuronal tumor cells of rats (B104). The chalone activity was found in the range of molecular weights between 10 000 and 100 000. The active part was of peptide nature, for proteolytic treatment destroyed the chalone activity.


Differentiation of mouse myeloid leukemia cells is inhibited by a factor from non-differentiating leukemia cells. Mouse myeloid leukemia cells (MI) were induced to differentiate by a factor(s) (D-factor) in ascitic fluid. An inhibitory activity (I-activity) for the induction of differentiation was present in conditioned medium and lysate of MI cells resistant to the D-factor. The I-activity was non-dialyzable, heat-labile and protease-sensitive. Most of the activity was recovered in the fraction precipitated with 30-50% saturated ammonium sulfate. The fraction inhibited induction of phagocytic activity, migrating activity and morphological changes in MI cells, which are typical properties of differentiated MI cells. Low levels of I-activity were detected in conditioned medium or lysate of MI cells sensitive to the D-factor. The resistant MI cells were sensitized to the D-factor by treatment with a low concentration (5-10 ng/ml) of actinomycin D. The I-activity in conditioned medium of actinomycin D-treated resistant cells decreased with development of sensitivity to the D-factor. These results suggest that production of the I-activity in the resistant cells is closely associated with resistance of the MI cells to the D-factor.


Toward an integrated approach to the institutionalized aged: an encounter with the medical model. The author describes the delivery of medical, psychiatric and social work services to residents of a facility for the aged. Although the Home had already benefited from the introduction of sound medical standards and the influence of an acute medical hospital, a satisfactory staff organization needed to be developed to answer the unique and individual needs of the aged residents. Specifically, the task was aimed at organizing services to implement a small unit concept of individual care for which the new facility was architecturally designed, to bring together those involved in patient care to coordinate services, to help the staff sustain a therapeutic attitude and to carry out specific therapeutic tasks.


A comparison of two response-reinforcement methods in pure-tone testing of the retarded. The purpose of this study was to determine whether a visual reward method or play audiometry is a better motivator in eliciting the lowest possible pure-tone thresholds from a group of Down's syndrome adults. Thresholds were obtained at 250, 500, 2000, 4000, and 6000 Hz for both procedures using a modified ascending method. The method of testing was constructed such that each of the two procedures served as a comparison for validity of the other. The level of significant difference between the play audiometry and the visual reward method was determined using the Sign Test. In each case, there appeared to be a significant difference between the two tests, but the difference was attributed to the order of administration of the tests and practice rather than by the tests themselves. The data obtained suggests that more accurate auditory thresholds may be determined by administering two pure-tone tests and accepting the second as most valid.


Specificity of nucleoside transport in Neurospora crassa. The specificity of nucleoside uptake in germinating conidia of Neurospora crassa was investigated by examining the kinetics of [2-14C]uridine and [8-14C]-adenosine uptake in the wild-type, ad-8, and ud-1 pyr-1 strains. The results obtained strongly indicate that nucleoside transport in N. crassa is mediated solely by a general transport system which accepts both purine and pyrimidine nucleosides. Studies directed at characterizing the specificity of the transport system indicate that general structural features of the nucleoside which enhance its efficiency in binding to the transport system include: (i) a purine or pyrimidine as the heterocyclic ring, (ii) an unfunctionalized ribose or 2'-deoxyribose as the sugar unit, (iii) a beta-configuration about the anomeric carbon, (iv) the absence of substituents at C8 in the purine series and at C5 and C6 in the pyrimidine series, (v) the presence of a C5-C6 double bond in the pyrimidine series, and (vi) the absence of a charge on the heterocyclic ring.


Isolation and characterization of glycerol-3-phosphate dehydrogenase-defective mutants of Neurospora crassa. Three glycerol-nonutilizing mutants deficient in the mitochondrial glycerol-3-phosphate (G3P) dehydrogenase (EC 1.1.99.5) were isolated from inl(ts) derivatives of Neurospora crassa following inositolless death at elevated temperatures on minimal glycerol medium. These mutants failed to grow on glycerol as a sole carbon source, but could grow on acetate, glucose, or mannitol media and were female fertile in genetic crosses, thereby distinguishing them from the previously reported polyol-protoperithecial defective Neurospora mutants. In addition, these glp mutants exhibited a distinct morphological alteration during vegetative growth on sucrose slants and colonial growth on sorbose-containing semicomplete medium. The glp-2 locus was assigned a location between arg-5 and nuc-2 on chromosome IIR on the basis of two-factor crosses and by duplication coverage by insertional translocation ALS176, but not NM177. All mutations were allelic as judged from the absence of both complementation in forced heterokaryons and genetic recombination among glp-2 mutations. The reversion frequency of all three mutations was less than 10(10), indicating probable deletions in these strains. No G3P dehydrogenase activity could be detected in either cytosolic or mitochondrial extracts from mutant strains grown on glycerol, glucose, or galactose media. These results suggest that the glp-2 locus may be the structural gene for both the cytosolic and mitochondrial forms of G3P dehydrogenase or for a cytosolic precursor of the mitochondrial G3P dehydrogenase. The defect is specific for the G3P dehydrogenase since normal activities of the mitochondrial cytochrome oxidase and succinate dehydrogenase and the cytosolic glycerol dehydrogenase and dihydroxyacetone phosphate reductase are detected in mutant extracts. During attempted growth of glp-2 mutants on glycerol media, there was an accumulation of G3P in culture filtrates, a reduction in the mycelial growth rate, and a decreased level of glycerokinase induction.


Cell wall composition and associated properties of methicillin-resistant Staphylococcus aureus strains. Methicillin-resistant (MR) Staphylococcus aureus strains have previously been reported to be deficient in surface negative charge; this has been correlated with methicillin resistance and ascribed to a deficiency of teichoic acid at the cell surface (A. W. Hill and A. M. James, Microbios 6:157-167, 1972). Teichoic acid was present in walls of MR organisms as revealed by appreciable phosphate levels and detection of ribitol residues. Phosphate levels in walls from five MR strains (0.54 to 0.77 mumol/mg of wall) were lower than in three unrelated methicillin-sensitive (MS) strains (0.86 to 1.0 mumol/mg of wall). However, two MS strains derived from two of the MR strains had wall phosphate levels very similar to those of the MR strains. No evidence for unusual wall polymers was found. Simple deficiency of wall teichoic acid does not result in methicillin resistance since an independently isolated teichoic acid-deficient strain (0.1 mumol of phosphate per mg of wall) was not methicillin resistant. In studies of biological properties possibly related to wall teichoic acid, it was discovered that walls isolated from MR organisms grown in the presence of methicillin autolyzed more rapidly than those isolated from organisms grown in the absence of the drug. Since methicillin resistance is enhanced by NaCl and suppressed by ethylenediaminetetraacetate, the effects of these compounds on autolysis of isolated walls were studied. NaCl (1.0 M) and ethylenediaminetetraacetate (1.0 mM) inhibited the autolysis of walls isolated from MR and MS strains. An MR strain bound phage 47, 52A, and 3A only slightly less well than their respective propagating strains.


Rapid purification of canine cardiac sarcoplasmic reticulum Ca2+-ATPase. A pure, enzymatically active Ca2+-dependent adenosine triphosphatase (Ca2+-ATPase) has been isolated from canine ventricular sarcoplasmic reticulum. In contrast to that derived from skeletal muscle, the Ca2+-ATPase from cardiac sarcoplasmic reticulum was more active when solubilization and subsequent purification took place in the presence of its substrates, Ca2+ and ATP. Cholate- or deoxycholate-solubilized Ca2+-ATPase is recovered following rapid glycerol dilution and centrifugation. The Ca2+-ATPase is stable and possesses hydrolytic capacities up to 4 mumol/mg/min. Sodium dodecyl sulfate-polyacrylamide gels reveal the presence of one protein in the range of 95,000 to 100,000 daltons. This method also yields purified Ca2+-ATPase from fast skeletal muscle of similar activities to those reported by other laboratories.


A comparative study of the myosin light chain kinases from myoblast and muscle sources. Studies on the kinases from proliferative rat myoblasts in culture, rat thigh muscle, and rabbit skeletal muscle. Myosin light chain kinases have been isolated from rat thigh and rabbit skeletal muscle and cultured rat myoblasts. From these preparations, two types of kinases can be distinguished: calcium-dependent and calcium-independent. Both types of kinases can phosphorylate isolated P-light chains of myosin from several sources (skeletal muscle, cardiac muscle, and platelet). Data are shown which support the phosphorylation of the same site on the non-muscle P-light chains by both types of kinases. The rates of these reactins are, however, different for the two types of kinases. Kinetic analysis of the myoblast kinase shows differing affinities for various P-light chains (non-muscle greater than cardiac greater than skeletal). In the proliferative rat myoblast, phosphorylation of myosin is a prerequisite for actin activation of the myosin ATPase activity.


Biochemical and structural characterization of actin from Dictyostelium discoideum. Actin has been purified from amoebae of Dictyostelium discoideum by a procedure which is notable in that proteolysis has been diminished to undetectable levels and "selective" purification steps have been avoided. The overall yield of this procedure is 5- to 10- fold greater than that of a previous report (Spudich, J. A. (1974) J. Biol. Chem. 249, 6013-6020). The detailed biochemical and structural properties of this new preparation (preparation B) have been compared to those of Dictyostelium actin prepared by the previous procedure (preparation A) as well as to rabbit skeletal muscle actin. Preparation B actin is similar to muscle actin in its molecular weight, ability to activate myosin, filament structure, and polymerization properties. Preparation B actin has the same molecular weight and isoelectric point as preparation A actin, which is more acidic than that of skeletal muscle actin. However, preparation B actin and muscle actin form longer filaments than preparation A actin, as judged by viscometry and electron microscopy.


Proximal femoral resection-interposition arthroplasty. In the severely retarded, multiply handicapped child who has a dislocated hip and severe adduction of the lower extremity, the deformity often interferes with perineal hygiene, nursing care, and positioning in bed and in a wheelchair. In twelve such patients we did an extensive resection of the proximal part of the femur, down to below the lesser trochanter, and constructed a capsular flap across the acetabulum. The quadriceps muscle was sutured around the resected end of the femur. This one-stage, uncomplicated operation allowed our patients to sit confortably and nursing care was made easy. In contrast, three patients who had single resection of the femoral head and neck had recurrence of deformity and pain.