Critical evaluation of prosthetic materials in repair of abdominal wall hernias: new criteria of tolerance and resistance. Six different materials (3 mesh and 3 cloth) commonly used for the repair of abdominal wall hernias were evaluated in 180 rats after implantation through musculature and peritoneum. Serial macroscopic and bacteriologic investigations were done, and bursting strength of the wound was determined in all groups at intervals with an original device. Histologic criteria were used to characterize the resistance of the wound and the tolerance of the host to the foreign material. Statistical analysis of the results demonstrated: (1) after postoperative day 15, the resistance of the wound was similar for each material tested; (2) during this early period mesh materials exhibited more resistance to bursting pressures than cloth materials; (3) the incorporation of mesh material was constant, whereas encystment or extrusion was always observed after implantation of cloth material; (4) no infection occurred with mesh material, but significant bacteria were found in 18 per cent of cloth material implantations; (5) the extent of the cellular reaction, the enumeration of giant, inflammatory, and fibroblastic cells showed the superiority of mesh materials; and (6) the ratio of fibroblasts to inflammatory cells reflected closely the mechanical resistance and tolerance of the foreign material.


Effects of denervation and tenotomy on the gastrocnemius muscle in the frog: a histologic and histochemical study. The effects of denervation on the gastrocnemius muscle of the frog were studied by histologic and histochemical methods. Thirteen Rana pipiens underwent unilateral sciatic neurotomy and were sacrificed weekly as long as 46 days. Of the three normal populations of muscle fibers, the small fibers underwent atrophy, the intermediate sized fibers remained unchanged in size, and the large fibers either did not change or underwent hypertrophy between 21 and 46 days. Necrosis of muscle fibers did not occur. Histochemical stains showed persistence of the normal pattern after denervation. The small fibers continued to have a high concentration of both oxidative and glycolytic enzyme activity (NADH-TR, SDH, phosphorylase), and the large fibers continued to have a low concentration of these enzymes. Depletion of glycogen stores was seen with PAS. Hypertrophic muscle fibers had mostly subsarcolemmal nuclei and few internal nuclei, suggesting that they may be physiologically tonic rather than twitch fibers. Achilles tenotomy at the time of denervation prevented the hypertrophy of large fibers. Abnormal inclusions have been demonstrated in mammalian muscle following tenotomy alone, but were not seen in the frog.


Paternal age effect in Down's syndrome.

Paternal age effect in Down's syndrome. Increasing incidence of Down's syndrome with advancing paternal age for given maternal age has been demonstrated. Comparisons are made between an almost complete Down's syndrome sample from the Copenhagen Metropolitan Area and a randomly selected sample of births from the same area and the same time period. Men above 55 years have a significantly increased risk of getting children with Down's syndrome.


Statistical methods for detecting a moderate paternal age effect on incidence of disorder when a maternal one is present. A statistical method is developed for detecting a moderate effect on the incidence of a disorder caused by advancing age of one parent when it is known that advancing age of the other parent is of great aetiological importance. The method is a conditional test procedure given the parental ages, therefore no assumptions about the parental age distributions have to be made. The method is applied on Danish material on Down's syndrome, for which a paternal age effect is demonstrated. Methods used in some well-known previous investigations have been discussed. Several of them, e.g. the classical one of Penrose (1933), could hardly detect any paternal age effect in Down's syndrome on the available data, because these methods are heavily affected by certain fertility patterns not recognized previously.


[Myocardium in valvular cardiopathies. Ultrastructural and histoenzymological changes. Functional correlations]. The morphologic and hemodynamic datas are compared in 24 patients with mitroaortic valvular cardiopathy. The morphologic study was carried out on left ventricular biopsies and consisted first of all in ultrastructural analysis. Thus, 5 degrees of alterations were determined with regard to myofibrillar and, accessorily, to mitochondrial changes. The microscopic data, especially rarefaction of the Z material on semifine sections stained with Toluidine Blue and the demonstration of phospholipids by staining with acid hematein, are consistent with the ultrastructural gradient. Also, in cases with the most advanced electronic gradient, histoenzymology revealed important anomalies in the activity of the myofibrillar ATPase and of the S.D.H. The correlation between the so-established morphologic gradient and the preoperative hemodynamic data proved satisfying in the majority of cases which points out the interest of the morphologic tests for the appreciation of the functional value of the myocardium.


[Immune complex disease of Swan and nude mice]. Swan and Nude mice with antinuclear antibodies (AN Ab) show early deposits of Ig and C in the mesangium and the basement membrane of glomerulus, marking an immune complexes disease. We have studied the Ig class of these immune complexes after acid elution and by direct immunofluorescence with suitable conjugates. In the Swan mice, fixed Ig are chiefly IgM and IgA, but AN Ab in the eluate kidney are principally IgG2 and IgM as the circulating AN Ab. In these mice the Ag of the fixed immun complexes are AN and murine leukemia virus Ag. In the Nude mice, fixed Ab are IgM and IgG2 and in part AN Ab. The circulating AN Ab and the AN Ab of eluates are also IgM and IgG2.


[Functional activity of cytolytic T lymphocytes in the rejection of tumor target cells]. Sensitized T lymphocytes can specifically destroy in vivo target cells with which they enter into direct contact. This suggests an important role for cytolytic T lymphocytes (CTL) in the destruction of grafts and tumours. Quantitative methods allow the assay of CTL and of CTL memory cells which appear in lymphoid tissues, blood, grafts and tumours during rejection. The formation of CTL and CTL memory cells can also be induced in vitro, and the functional activity of such cells has been demonstrated in transfer experiments. In the present report, results of experiments will be discussed which were aimed at the analysis of the characteristics of memory cells, of the functional activity in vivo of CTL induced in secondary mixed leucocyte cultures (MLC), and of the specificity of CTL formed in response to tumours induced by the Moloney sarcoma virus (MSV). Our results show that there exists a qualitative difference between the original precursor of the CTL and its memory cell, and that the transfer by the intravenous route of CTL induced in secondary MLC protects irradiated mice effectively and specifically against allogeneic tumour growth. In the MSV system, studies of the specificity of CTL show that syngeneic tumour cells are lysed preferentially (and that allogeneic MSV induced tumour cells are not lysed), indicating a genetic restriction in their activity.


[New method of diagnosis of drug hypersensitivity of the delayed type--the scarificaiton-film test]. An original method for diagnosis of drug allergy, i.e. a scarification-film test (SFT) in 2 variants has been worked out for the first itme on the basis of examination of 241 persons and is recommended for use. 10 mm parallel scarifications with intervals of 3-4 mm between them were made on the epidermis of the antibrachial flexor and the following agents were applied to them: 1) a film-forming aerosol composition with medicines (tetracycline)--the 1st variant of SFT. (2) powder or solutions of medicines (antibiotics, sulfanylamides, vitamins, etc.) covered with a film-forming ocmposition without any medicine--the 2nd variant of SFT, 3) film-forming aerosol composition without any medicine-control of both variants out of 46 patients with professional dermatosis and in 10 (7.8 per cent) out of 127 practically healthy persons with latent sensitization (workers of pharmaceutical factories) being in contact with tetracycline. Hypersensitivity of the delay type to benzylpenicillin, streptomycin, sulfanylamides and other medicines was found in 54 79.4 per cent) out of 68 patients with drug allergy of professional and non-professional etiology. The method was shown to be highly sensitive, handy, simple and safe, which provided its recommendation for the use in wide clinical practice for diagnosis of drug allergy of the retarded type which developed because of drug use or professional contact.


[Olemorphocycline treatment of calculous and noncalculous pyelonephritis]. 40 patients suffering from calculous and non-calculous pyelonephrits were treated with olemorphocycline administered intravenously in a dose of 250 00 Units 2 times a day for 5--7 days. The clinical effect was observed in 62.5 per cent of the cases. In 54.7 per cent of the cases the isolates were resistant to oleandomycin and morphocycline. The kidney tissue and urine from the kidney pelvis taken during surgical operations of 20 patients were plated out. Staphylocci were most often detected in the platings of the kidney tissue. The microflora of the urinary bladder before the treatment and that of the urine from the kidney pelvis and the kidney tissue taken during the surgical operation was mainly ident


[Lymphocyte impairment in aging].

[Lymphocyte impairment in aging]. In vitro stimulation of peripheral blood lymphocytes  by T mitogens and in MLC is reduced in aging. As the number of T lymphocytes and their antigenic power are in the normal range, a functional T-cell defect is anticipated in high age.


Acne fulminans: investigation of acute febrile ulcerative acne. Eight cases of acne fulminans (acute febrile ulcerative acne) are compared with 13 previously reported cases. This rare disorder of male teenage patients is characterized by the sudden appearance of highly inflammatory, tender, ulcerative and crusted lesions on the back, chest, and face: it is one of the most scarring acute dermatologic disorders of young patients. At the onset of the disease, all patients showed febrile temperatures and marked leukocytosis; other systemic symptoms occurred with varying frequency. Polyarthralgia was noted in half of our series. Investigations of potential trigger mechanisms (eg, septisemia due to Corynebacterium acnes, immunologic defects, altered polymorphonuclear leukocyte function, Shwartzman phenomenon) did not yield any clues regarding the pathogenesis of this disease. Clinical and laboratory findings suggest that acne fulminans is not directly related to acne conglobata.


Hidrotic ectodermal dysplasia: study of a large Chinese pedigree. Hidrotic ectodermaldysplasia was found, to our knowledge, for the first time in a Chinese family in Malaysia, and it affected 15 members in five generations. The disease, which is transmitted as a non-sex-linked autosomal dominant trait, presumably originated from southern China. All 15 members had the typical nail, hair, and skin lesions, and we observed three different types of nail defects. Scalp alopeica was more extensive in the female members while keratoderma of the palms and soles was more notable in the male members. The nail and skin lesions also became severer with age. Except for the infectious eczematoid dermatitis present in the propositus, none had other skin or systemic disorders. All were relatively healthy and had normal life expectancies;


Metronidazole ("Flagyl") in dracunculiasis: a double blind study. Two-hundred and twenty-nine cases dracunculiasis were selected for a double-blind trial of metronidazole against placebo. A cure rate of 85% was observed with metronidazole. A dosage of 400 mg metronidazole three times daily for 10-20 days appears suitable. Even with secondarily infected lesions, it was unnecessary to administer any other chemotherapeutic agent. In most cases symptomatic relief, especially of pain and pruritus, was obtained within two weeks. In patients with only subcutaneous worms, metronidazole did not apparently prevent the development of lesions and seemed to stimulate the worm to emerge quickly, with resultant less severe lesions. Complete cure was delayed in patients with multiple lesions, where worms reached the emergence state at different times. There did not appear to be any direct relationship between severity of the disease and response to metronidazole. If the worm was broken during treatment with metronidazole, no abscess formed nor was there any local inflammation. Metronidazole was very well tolerated even when administered for 20 to 25 days. No serious side-effects or toxic effects were observed.


Prefabricated Dacron baffle for use in correction of transposition of the great arteries. Interatrial venous transposition for correction of transposition of the great arteries (TGA) is facilitated by using a premolded Dacron baffle. Fine-mesh Dacron is easily fashioned to the desired shape in a metallic brass mold at 200 degrees C. The resultant baffle material can be autoclaved without losing its shape or altering the fabric characteristics of flexibility and porosity. This premolded baffle has been used in our last 7 patients with TGA over the past two years without complication, and the use of Dacron in 13 survivors over the past seven years has shown no pulmonary or venous obstructions related to the material.


Studies of urticaria and acute serum sickness with the C1q precipitin test. The C1q precipitin test was performed in serum samples from five groups of patients: (1) 20 patients with acomplementemic systemic lupus erythematosus glomerulonephritis (SLE), (2) 2 patients with serum sickness due to the administration of horse serum, (3) 2 patients with serum sickness preceding hepatitis B, (4) 50 patients with chronic urticaria, and (5) 30 normal controls. Positive C1q precipitin tests were found in all patients with SLE and the four cases of serum sickness. Positive tests correlated with depressed serum complement (C3 and C4) levels and were found only in the early phase of serum sickness. Urticaria patients uniformly had negative C1q precipitin tests and normal serum complement levels.


[Left atrial myxoma: a surgical emergency. Apropos of 2 cases]. This study reports on two more cases of left atrial myxoma. They were two women with embolic manifestation for several years and in whom the diagnosis was not clinically considered. It was made by echocardiography. Emergency surgery was applied in both cases one of whom without hemodynamic study. The presence of a recent brain embolism did not seem to us to be a contraindication to this surgery. In both cases the tumoral pedicle was implanted on the posterior wall of the left atrium, an unusual location.


[Cardiovascular signs in drepanocytosis. Apropos of 12 cases]. Sickle cell disease is the commonest of the haemoglobinopathies. The pathophysiology consists on the one hand of chronic haemolytic anaemia whose course is punctuated by acute crises with pain, fever and severe haemolysis, and on the other of a tendency to microthrombotic accidents. Of all the very varied clinical features of the disease, cardiac signs are both common and variable. The authors report a series of 92 cases, and compare their findings with those in the literature.


Pathways of D-fructose and D-glucose catabolism in marine species of Alcaligenes, Pseudomonas marina, and Alteromonas communis. Cell-free extracts of D-fructose grown cells of marine species of Alcaligenes as well as Pseudomonas marina contained an activity which catalyzed a P-enolpyruvate-dependent phosphorylation of D-fructose in the 1-position as well as activities of the following enzymes: 1-P-fructokinase, fructose-1,6-P2 aldolase, PPi-dependent 6-P-fructokinase, fructokinase, glucokinase, P-hexose isomerase, glucose-6-P dehydrogenase, 6-P-gluconate dehydrase, and 2-keto-3-deoxy-6-P-gluconate aldolase. The presence of these enzyme activites would allow D-fructose to be degraded by the Embden-Meyerhof pathway and/or the Entner-Doudoroff pathway. In cell-free extracts of D-glucose grown cells, the activity catalyzing a P-enolpyruvate-dependent phosphorylation of D-fructose as well as 1-P-fructokinase activity were reduced or absent while the remaining enzymes were present at levels similar to those found in D-fructose grown cells. Radiolabeling experiments suggested that both D-fructose and D-glucose were utilized primarily via the Entner-Doudoroff pathway. Alteromonas communis, a marine species lacking 1-P-fructokinase and the PPi-dependent 6-P-fructokinase, contained all the enzyme activites necessary for the catabolism of D-fructose and D-glucose by the Entner-Doudoroff pathway; the involvement of this pathway was also consitent with the results of the radiolabeling experiments.


Huntington disease: early indentification by H reflex testing. This investigation was undertaken to determine the potential use of the H reflex of Hoffmann as a means of early detection of Huntington disease. Three groups of subjects were tested: 9 having Huntington disease, 8 first-generation offspring "at risk" of Huntington disease, and 30 normal persons as controls. The deep peroneal nerve was stimulated by a monopolar needle electrode, and the muscle potentials of the anterior tibialis and the extensor digitorum brevis were recorded with surface elctrodes. The action potentials were examined for the direct M response and the presence or absence of an H reflex response. The H reflex response was found in the anterior tibialis muscle, at least unilaterally if not bilaterally, in eight of nine subjects with Huntington disease and in five of eight persons at risk. The H reflex was present in 1 of 30 controls unilaterally. This significant difference between the patients with Huntington disease and at risk subjects compared to the control group, indicates that H reflex testing may be useful for the early identification of at risk individuals who will ultimately develop Huntington disease. This would be most helpful in family counseling terms, and also may aid in further understanding of this complex diseases.


Bus use by disabled arthritics: functional requirements. Twenty-five rheumatoid arthritic patients were studied to determine the extent of independence and functional requirements necessary to use public bus transportation in Philadelphia. To measure the extent of independence, a functional assessment scale was developed that evaluates 19 bus activities, each according to a four-point scale. Of the 11 subjects who stated that they used the public bus prior to admission, only 4 were found to be able to use the public bus independently at the time of evaluation. Ascending, descending and motion activities provided considerable difficulty for the sample population. Functional requirements of boarding and disembarking the bus are described in detail.


[Morphogenesis of amyloidosis].

[Morphogenesis of amyloidosis]. Basing on the literature data and findings of their own investigations with the use of sophisticated methods of morphological analysis, the authors showed that morphogenesis of experimental amyloidosis comprised the following links: 1) cellular transformations of the reticulo-endothelial system with formation of a clone of amyloidoblasts; 2) synthesis by these cells of protein of amyloid fibrils; 3) aggregation of fibrils with formation of a "carcass" of amyloid substance; 4) combination of aggregated fibrils with proteins and glucoproteids of plasma and also with acid mucopolysaccharides of the tissue and formation of a complex glucoproteid -- amyloid. The sources of amyloblasts in various organs were precursors of different cells of mesenchymal nature: in the spleen -- reticular and endothelial cells; in the liver -- Kupffer's cells; in the kidney -- mesangial and endothelial cells. In the course of formation of amyloid fibrils, the latter being anomalous protein of the body, a "competition" started between the synthesis of fibrillar protein and its resorption (amyloidoclasia). It completed in favour of the synthesis, which may be explained by the development of immunological tolerance to amyloid protein. At the final stage of amyloidogenesis of particular importance was an increase in the tissue-vascular permeability, which ensured the extracellular formation of amyloid substance and inclusion therein of hematogenic "additions" (fibrin', immune complexes, etc.).


[Work of the centralized pathoanatomical department of the Cheliabinsk district clinical hospital]. The article is dedicated to the organization of work of the centralized pathologoanatomic department of the Cheliabinsk  regional clinical hospital which offers its service to 53 curative institutions with a total number of 7965 beds. A required material base has been created, and the department is equipped with modern apparatus and appliences. Staff members of the Department include 13 medical pathoanatomists, 21 laboratory technicians-histologists, and 15 aid men; the available personnel makes it possible to organize in the most rational way the work of every staff member, and to put into practice the methods of scientific organization of labour. The department has become a pathologoanatomic centre where an important organizational-methodological work is being carried out on a large scale.


[Pigmented epitheliomas of the eyelids]. Pigmented epitheliomas are generally basocellular epitheliomas tatooed with melanin. Certain clinical characteristics differentiate them from true melanomas. They respond to radiotherapy.


[Opsoclonus and associated neurologic disorders. General review and commentary on a case]. A case report on a young man who developed after cranial trauma a typical opsoclonus and neurological symptoms dominated by left sensory hemiparesis of the thalamic type; this deficit affected all sensory modalities and involved the whole of the left side, with disturbances of motility caused by impairment of sensation. A general review of opsoclonus is presented and supported by various neurological tables. The discussion on aetiology takes into account the frequent benign causes (encephalitic syndromes, ataxic forms of poliomyelitis), but also the possibility of serious lesions developing in the cerebellum and cerebrum; and finally there remain a number of cases of unknown aetiology. Clinical observations of opsoclonus are rare, and anatomical data is still less; this is why physiopathological interpretations, the object of interesting discussions, are only reached with wisdom.


[Color discrimination in heterozygotes]. By the help of Rabkin polychromatics tables and of the AN-59 anomaloscope it is studied the chromatic eye sight at 33 heterozygots. The used tests have permitted the identification of the different types of heterozygots: heterozygots which transmit and manifest only one chromatic anomaly, double heterozygots which manifest clinical one or both anomalies and unmanifested clinical heterozygots. The study of the chromatic eyesight at men from the same family has confirmed the clinical diagnosis of the heterozygots type in every case.


[The use of freeze-etching in the ultrastructural study of the cornea]. The freeze-etching and freeze-fracturing techniques allow a good examination of fixed and unfixed tissues by the mean of transmission electron microscope. As compared with usual techniques of fixed and embedded tissues we can observe the ultrastructural characteristics of the rabbit cornea: epithelium, stroma and endothelium. This technique is the best one to give a good view of the intercellular junctions. 1) On the epithelial layers we describe intracytoplasmic bunches of filaments and numerous desmosomes. After freeze-fracturing the desmosomes appeared to be formed of aggregated particles on fractured faces. These particles are possibly the point of attachment of intracytoplasmic tonofibrils.


[Diagnostic and therapeutic use of cyclopentolate cycloplegia]. 0,5 p. 100 Cyclopentolate is an important short-acting cycloplegic and mydriatic preparation. It has the same effect on accommodation as the prolonged instillation of Atropine. Nevertheless it must be remembered that it is not a perfect cycloplegic; just as much as with Atropine resistance is frequently found in young children, in subjects with brown irides, in pigmented races, and above all in cases where there is a tonic spasm of accommodation producing functional signs or oculomotor imbalance.


[Ocular microsurgery. Contemporary ideas]. Microsurgery in ophthalmic practice consists of performing certain operations on the eye with the aid of a powerful binocular telescopic system which gives a greatly magnified view of the ocular structures. The equipment in use has improved progressively since 1953. The classical operations have been improved and new surgical techniques are now possible.


[Eyelid magnets for facial paralysis].

[Eyelid magnets for facial paralysis]. The author presents his experience in the use of lid magnets. The positioning is delicate but tolerance is good and the aesthetic and functional result satisfactory. The essential indication is the development of corneal problems overnight from defective occlusion.


[Residual astigmatism after transfixing keratoplasty in keratoconus]. The authors made a statistical study of the refraction and corneal topography in 21 patients who had penetrating grafts for keratoconus performed by the same surgeon one year or more previously. They used various criteria among which the following were important: -- the importance of Javal ophthalmometry to determine the astigmatism for prescription; -- good visual results; -- the recording of significant corneal deformities during the topographical studies.


[Ultrastructural study of the oculomotor muscle in 2 familial cases of Steinert's disease]. A biopsy specimen of an extra-ocular muscle from a brother and sister affected by typical familial Steinert's disease was examined by electron microscopy. Considerable disorganisation of the contractile element was observed with a gross accumulation of abnormal mitochondria. This was a very different appearance to the findings observed in skeletal muscle in Steinert's disease and much closer to the findings in skeletal muscle in certain 'ocular' myopathies. One cannot fail to be impressed by the discordance between the appearance of the muscular disorganisation and the rarity of clinical oculomotor disturbances in this disease.


[Closed angle glaucoma induced by phospholine iodide and epinephrine in a case of open angle glaucoma]. A patient suffering from open angle glaucoma was treated with topical phospholine iodide and epinephrine. Before treatment the anterior chamber was deep and the angle wide open in both eyes. Two weeks after beginning treatment there was an acute elevation of pressure to 52 mm hg in the right eye accompanied by myopia, shallowing of the anterior chamber and closure of the angle. These findings were dramatically reversed by the instillation of one drop of tropicamide and one drop of cyclopentolate. Simultaneous contraction of the dilator and sphincter muscles of the iris in this case probably caused pupillary block and closure of the anterior chamber angle, in spite of the initially wide open angle. Phospholine iodide may also have caused a spasm of the ciliary muscle and a forward movement of the lens, in a manner similar to that thought to occur in malignant glaucoma. Strong miotics together with epinephrine should be used cautiously in the treatment of glaucoma even when the initial examination shows a wide open angle.


Effect of heptane treatment on the response of sarcoplasmic reticulum preparations to phosphate. The calcium-stimulated (extra) ATPase and calcium uptake activities of sarcoplasmic reticulum (SR) preparations treated with aqueous heptane mixtures were compared with those of untreated SR, and with those of SR treated with aqueous ether. Both treatments altered the kinetic behaviour of the extra ATPase, the Lineweaver-Burk plot being changed from its normal non-linear shape to a straight line. Kinetic constants, Vmax, Km for ATP and Ki for phosphate, were measured. The extra ATPase activity of heptane-treated SR was inhibited by phosphate as was that of ether-treated SR, to a lesser extent. The magnitude of this inhibition by phosphate was found to be considerably less than the degree of stimulation of the extra ATPase activity of untreated SR caused by phosphate through its calcium-precipitating action. The steady-state concentrations of the phosphoryl-enzyme intermediates were measured and together with the Km and Ki values they indicate that the bidning of ATP to heptane-treated SR is weaker than it is to untreated SR, and that phosphate is an efficient competitor for the binding sites.


[Episcleral indentation by lyophilized organic implants in surgery of retinal detachment]. The purpose of this paper is to describe an original technique for surgical treatment of idiopathic retinal detachment. Since 1971, the authors use human organic implants (lyphoylised scléral tissue). These implants are included and tied in front of the retinal tears, to obtain a large and deep scléral indentation. There is neither immunological rejection nor local inflammation. The results of 455 cases of retinal detachment are reported and analysed. This operative procedure gives 80 p. 100 of good résults. At this day, 1000 patients has been operated with this surgical procedure, at the University Eye Clinic A. of Lyons.


[Phakoemulsification using the Girard apparatus. Experimental study of the effects on the corneal endothelium of the rabbit]. The purpose of this paper was to investigate the potential alterations of the corneal endothelium following phacofragmentation with the Girard Ultrasonic Phacofragmentation Unit. The study was done in rabbit corneas using the specular microscope, nitroblue tetrazoium staining, and scanning electron microscopy. The perfusion fluid was Plasmalyte 148 R (Travenol), a solution which has been previously evaluated in this type of operation. Two experiments were done: (1) The effects of ultra-sound and irrigation for a period of three minutes, and (2) The effect of phacofragmentation for an average time of ten minutes. Results of the first experiment showed no cell destruction, but increased permeability to nitroblue tetrazolium. The second study showed areas of isolated or confluent cell destruction probably caused by mechanical trauma due to instrumentation or by lens nucleus rubbing against the corneal endothelium. These lesions have been observed with other instruments in similar techniques of lens removal.


The epidemiology of hepatitis B in a residential institution for the mentally retarded. A longitudinal study carried out over 43 months in a residential home for the mentally retarded confirmed the marked propensity of individuals with Down's syndrome (DS) to develop chronic HB antigenaemia. This could not be accounted for by environmental factors, and a genetic basis is postulated. In addition, DS subjects appeared innately more susceptible to hepatitis B virus (HBV) infection, requiring one-third the patient-months of exposure of non-Down's subjects (ND) before showing evidence of infection. The reciprocal geometric mean titre of anti-HBS in DS subjects was 110 compared with 705 in ND subjects, suggesting a deficiency of humoral immunity in the DS subjects. Transmission of HBV was occurring by means other than therapeutic, prophylactic, dental or diagnostic procedures, in the institution, and possible modes of transmission are discussed. These were probably both parenteral and non-parenteral. Eighty-one per cent of HBV infections in the Home were subclinical.


Drug induced interstitial nephritis, hepatitis and exfoliative dermatitis. Acute interstitial nephritis associated with hepatitis, exfoliative dermatitis, fever and eosinophilia is uncommon. The syndrome has been described previously in association with phenindione administration, leptospirosis and heavy metal poisoning. Four cases are described, two of which were due to phenindione sensitivity. The other two patients had been exposed to a number of toxins including allopurinol, frusemide, chlorothiazide and methyldopa so that the exact aetiological agent is unclear. Interstitial nephritis should be considered as a cause of acute renal failure in patients with other features of drug hypersensitivity.


[Transmission of the anterior chamber pressure to the center of the vitreous body and to the posterior pole of the eye of the miniature swine. Measurement of isovolumetric and direct pressure]. Experimental variations of the anterior chamber pressure (P.I.C.) are transmitted at about 300 ms to the middle of the vitreous. The response time to a sudden experimental increase in P.I.C. to the centre of the vitreous is variable and depends essentially on the control intra-ocular pressure: the more it is raised the shorter the response time. The pressure in the centre of the vitreous (P.I.V.) has a pulsatile character probably due to transmission of arterial pulsation. The higher the intra-ocular pressure the greater the amplitude of these pulsations. Recording of P.I.V. close to the surface of the retina show abrupt falls despite the fact that the P.I.C. and systemic arterial pressure remain stable. These results are discussed in relationship to autoregulation of the retinal circulation and the physiopathology of glaucoma.


The role of immune complexes in the pathogenesis of disease. Circulating antigen-antibody complexes are incriminated in the pathogenesis of auto-immune and inflammatory disease, and more recently malignancy. Extensive knowledge of the immunopathological reactions has evolved from from the study of experimental serum sickness in animals and of the potential aetiological agents (e.g. viruses) from spontaneous immune complex diseases in animals. Numerous techniques, both direct and indirect, have now been described to identify immune complexes in serum, though no single technique will identify regularly immune complexes in all clinical situations, nor will it demonstrate the pathogenicity of the immune complex in a given patient. Human disorders with a definite immune complex basis (glomerulonephritis, systemic lupus erythematosus, rheumatoid arthritis) and others with a possible immune complex basis (e.g. cutaneous vasculitis, are presented. Management of immune complex disorders is based on removal of the initiating agent if known (e.g. infection, drug, malignancy) or the use of non-specific anti-inflammatory therapy. Specific immunotherapy, in practice and theory, is discussed.


The peritoneal complications of ventriculo-peritoneal shunts. One hundred and two patients with ventriculo-peritoneal shunts were studied. The aim of the study was to record the complications caused by the peritoneal end of the catheter and the frequency with which they occurred. Some of the complications involving tube migration are illustrated by case reports emphasizing the possible morbidity adn mortality that may be associated with this problem. Fity-one patients in the series had a complication; of these, 23 had only a mild temporary intestinal ileus following initial insertion of the shunt system. In the remaining 28 patients, the complications could have been significantly reduced by careful attention to operative detail and a closer follow-up of the patients following the shunt insertion.


[Acetylcholine in the bovine retina and the parasympathomimetic effect of pilocarpine]. Using titrations of acetylcholine (ACh) on guinea-pig ileum the authors have demonstrated that there is no significant statistical difference between the cholinergic activity of extracts of 150 mg of retinal tissue from pilocarpinised eyes (0.3 mug of ACh) and the activity of extracts from control eyes not subjected to pilocarpine (0.29 mug of ACh). The pilocarpine had been instilled into the bovine conjunctival sac at the rate of three drops of 3 p. 100 pilocarpine HCl ten times at intervals of three minutes. One hour after the last application the animals were killed and the eyes enucleated. The role of pilocarpine in the regulation of tissues in the posterior segment of the eye is discussed.


Optical properties and denaturation by guanidinium chloride and urea of the adenosine triphosphatase of Micrococcus lysodeikticus. A comparison of four molecular forms of the enzyme. 1. The fluorescence and circular dichroism of four homogeneous preparations of ATPase (adenosine triphosphatase) from Micrococcus lysodeikticus differing in molecular structure and enzymic properties were examined at pH 7.5 and 25 degrees. Emission was maximum at 325 and 335 nm and the relative intensities at these wavelengths may be used to characterize the different ATPase preparations. The circular-dichroism spectra exhibited negative extrema at 208 and 220 nm, and the relative value of the molar ellipticity at these wavelengths was also different for each molecular form of the enzyme. 2. The four preparations undergo two consecutive major unfolding transitions in guanidinium chloride (midpoints at 0.94 and 1.5 M denaturant), with concomitant destruction of the quaternary structure of the protein. A comparatively minor alteration in the ATPase structure also occurred in 0.05-0.2M-guanidine and led to complete inactivation of the enzyme. The inactivation and the first unfolding transition were reversible by dilution of the denaturant; the transition with midpoint at 1.5M-guanidine was irreversible. 3. Similar results were obtained in urea, except that the successive transitions had midpoints at concentrations of denaturant of 0.4, 2.0 and 4.5M. Low concentrations of urea caused a noticeable activation of the enzyme activity and alterations of the electrophoretic mobility of the ATPase. 4. A model is proposed in which one of the major subunits, alpha, is first dissociated and unfolded reversibly by the denaturants, followed by the irreversible unfolding and dissociation of the other major subunit, beta, from subunit delta and/or the components of relative mobility 1.0 in dodecyl sulphate/polyacrylamide-gel electrophoresis (rho).


Molecular conformation of sodium heparan sulphate in the condensed phase. By using the X-ray-diffraction results reported previously for sodium heparan sulphate, a twofold helical conformation with an axially projected disaccharide repeat (h) equal to 0.93 nm has been examined in detail. On the basis of a repeating sequence of 1,4-alpha-D-glucosamine and 1,4-beta-D-glucuronic acid, trial and stereochemically feasible molecular models were computer-generated. An optimum twofold helical conformation is proposed, incorporating stabilizing intra-chain hydrogen bonds across both glycosidic linkages.


The subunit structure of the arom multienzyme complex of Neurospora crassa. A possible pentafunctional polypeptide chain. A new procedure for the purification of the arom multienzyme complex from Neurospora crassa is presented. Important factors are the inactivation of proteinases by phenylmethanesulphonyl fluoride and the use of cellulose phosphate as an affinity adsorbent. A homogeneous enzyme, with a specific shikimate dehydrogenase activity of 70 units/mg of protein, is obtained in 25% yield. Polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate, combined with cross-linking studies using dimethyl suberimidate, suggest that the complex is composed of two subunits of molecular weight 165000. Glycerol-density-gradient centrifugation indicates a molecular weight for the intact complex of about 270000. Evidence for the effects of proteolysis, both during the preparation and on storage of the purified complex, is presented, and previous reports in the literature of the occurrence of multiple subunits are discussed in this light.


Mitochondrial adenosine triphosphatase of the fission yeast, Schizosaccharomyces pombe 972h-. Changes in activity and oligomycin-sensitivity during the cell cycle of catabolite-repressed and -de-repressed cells. 1. Changes in activity of ATPase (adenosine triphosphatase) during the cell cycle of Schizosaccharomyces pombe were analysed in cell-free extracts of cells harvested from different stages of growth of synchronous cultures and also after cell-cycle fractionation. 2. Oligomycin-sensitive ATPase oscillates in both glucose-repressed synchronous cultures and shows four maxima of activity approximately equally spaced through the cell cycle. The amplitude of the oscillations accounts for between 13 and 80% of the total activity at different times in the cell cycle. 3. Oligomycin sensitivity varies over a fourfold range at different stages of the cell cycle. 4. The periodicity of maximum oligomycin sensitivity is one-quarter of a cell cycle. 5. These results were confirmed for the first three-quarters of the cell cycle by cell-cycle fractionation. 6. In cells growing synchronously with glycerol, ATPase activity increases in a stepwise pattern, with two steps per cell cycle; the first of these occurs at 0.54 of the cell cycle and the second at 0.95. 7. These results are discussed in relation to previously obtained data on the development of mitochondrial activities during the cell cycle.


The adenosine triphosphatase-inhibitor content of bovine heart submitochondrial particles. Influence of the inhibitor on adenosine triphosphate-dependent reactions. 1. The activity of the ATPase (adenosine triphosphatase) of phosphorylating particles prepared by sonication of bovine heart mitochondria in the presence of MgCl2 and ATP is influenced by the isolation method for the mitochondria used in the preparation of particles. Type-I particles, made from mitochondria isolated in a medium lacking succinate, have a lower ATPase activity than to Type-II particles, which are prepared from mitochondria isolated in a medium containing succinate. 2. Centrifugation under appropriate energized conditions increases the ATPase activity of Type-I particles almost to that of the Type-II particles. The ATPase activity of Type-II particles was only slightly stimulated by this procedure. These data are interpreted as indicating a higher content of the ATPase-inhibitor protein in the Type-I particles. 3. A comparison was made of the ATP-driven enhancement of 8-anilinonaphthalene-1-sulphonate fluorescence and the exchange of the endogenous tightly bound nucleotides of the ATPase in Type-I and Type-II particles. The effect of exogenous inhibitor protein on both these reactions was also studied. 4. The time-scale on which the inhibitor protein can exchange between ATPase molecules is discussed.


[Fluorometric determination of glibornuride in plasma and serum using 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole]. Glibornuride, tolbutamide and gilbenclamide react on heating with 7-chloro-4-nitrobenzo-2-oxa-1.3-diazole (NBD-chloride) in amylacetate to form a highly fluorescent NBD-amine. A fluorometric determination of glibornuride is presented. Reaction conditions and the extraction of glibornuride from plasma and serum are described. The method is sensitive to measure drug levels after therapeutic doses. The detection limit is ca. 40 ng/ml and the recovery rate is ca. 100%. The coefficients of variation for the determination of drug levels of 1 to 0.1 mug/ml range from 4.1% to 12.1%.


Influence of dietary status and diabetes on aortic acyl-CoA hydrolase activity. We have postulated that the accelerated snythesis of cholesteryl ester in atherosclerotic microsomes may result in part from decreased acyl-CoA hydrolase activity in arterial tissue, because acyl-CoA is a common substrate for both reactions. We have now investigated the influence of nutritional status, type of diet, and diabetes on the acyl-CoA hydrolase activity of otherwise normal aortic microsomes. Fasting rabbits for 16 hr diminished the acyl-CoA hydrolase activity approximately 30%. The activity of this aortic microsomal enzyme in rats maintained on a high-carbohydrate diet for 5 weeks was comparable to the activity observed on a high fat (olive oil) diet. The type of fat in the diet influences the acyl-CoA hydrolase activity: oils containing 77% oleic acid (high-oleic safflower oil) and containing 70% linoleic acid (conventional safflower oil) lowered the aortic microsomal acyl-CoA hydrolase activity in comparison to a more saturated fat (cocoa butter). Aortic preparations of rats made diabetic by streptozotocin exhibited higher acyl-CoA hydrolase activity than the normal. The results show that conditions associated with human atherogenesis (diabetes and saturated fat diet) increase rather than suppress the activity of this arterial enzyme in normal arterial tissues of the rat.


Hyperlipidaemia in patients with Down's syndrome. Plasma lipid levels were measured in 20 mongoloid and 16 non-mongoloid mentally retarded subjects. Significant elevations of plasma triglyceride levels were found in patients with Down's syndrome compared with mentally retarded controls. However, no significant difference was found in plasma cholesterol, phospholipid and free fatty acid levels between the mongoloids and control subjects.


The left ventricle in congenital isolated pulmonary valve stenosis. A morphological study. The wall thickness of the left ventricle was measured in the hearts of 16 children and 2 adults who died of congenital pulmonary valve stenosis. A right-to-left shunt through a patent foramen ovale had existed in 12 cases and was excluded in the other 6. The thickness of the left ventricular wall and the interventricular septum was increased in the majority of hearts, especially in the older patients. There was a good correlation between wall thickness and histological appearance of the left ventricular myocardium; in 2 cases there was also evidence of left ventricular myocardial fibrosis. The coronary arteries appeared normal in all cases. A direct interaction between the ventricles was thought to be a possible mechanism.


Further evidence of response by leukaemia patients in remission to antigen(s) related to acute myelogenous leukaemia. Fifteen patients with acute myelogenous leukaemia were studied to determine if their remission blood leucocytes could be stimulated into taking up [3H] thymidine after in vitro culture with their own cryo-preserved irradiated AML leukaemia cells. In 6/15 patients it was possible to show autologous recognition and equal recognition of their stored leukaemia cells, even when they had previously been maintained in in vitro proliferative cultures in liquid suspension and undergoing myeloid maturation for one week. After in vitro proliferative culture, 4 populations of leukaemia cells produced material in the supernatant media between 3 and 7 days capable of inducing [3H] thymidine uptake in autologous (2 pts, 5 supernatants) and allogeneic (2 pts, 2 supernatants) AML remission lymphocytes, but not in normal donor lymphocytes. The relevance of these observations to tumour-associated AML antigen is discussed.


Fixed drug eruption: ultrastructural study of dyskeratotic cells. Electron microscopic studies have been carried out on three cases of fixed drug eruption, with particular regard to the dyskeratotic cells. The authors have tried to show the sequence of events leading from a normal basal keratinocyte to a dyskeratotic body. Ribosomes were first increased in number; then the tonofibrillar system looked thicker, cytoplasmic organelles degenerated and numerous melanosomes appeared. Many dyskeratotic bodies were later found in epidermal macrophages and in the intercellular space. Several intra-cytoplasmic desmosomes were found in the dyskeratotic cells and their evolution is discussed. The increased number of melanosomes seen in the dyskeratotic bodies could be due to injury to epidermal lysosomal catabolism or it could be due to an apparent increase in melanosome numbers, within cells whose volume has been reduced.


Serum zinc and retinol-binding protein in acne. The serum levels of zinc and retinol-binding protein (RBP) have been determined in 173 patients with acne and compared with those of a control group. The RBP is a specific transport protein and its level in plasma reflects the amount of vitamin A available to the tissues. Patients with severe acne were found to have lower levels of RBP than either patients with mild acne or healthy subjects of the same age. In the case of males with severe acne, the mean serum zinc level was significantly lower than that of the control group. No such difference was observed for girls. The observed condition of low levels of zinc and vitamin A in the serum of patients with severe acne may provide a rationale for the clinically good effect of oral zinc treatment.


An acne-free naevus.

An acne-free naevus. Investigation of rare disorders may sometimes help in the understanding of common problems. We describe a patient with an unusual and previously unreported acne-free naevus occurring in an area severely affected by acne. Investigation revealed smaller sebaceous glands, a reduced sebum excretion rate, decreased number of Corynebacterium acnes in the pilosebaceous ducts and a reduced conversion of testosterone to 5 alphadihydrotestosterone in the areas which were free of acne compared with acne-bearing skin. The results of these investigations support some of the suggested mechanisms responsible for the development of acne.


Myosin in cultured human endothelial cells. Myosin was isolated from cultured human endothelial cells by extraction with 0.6 M KCl and chromatography on Sepharose 4B. The extracted endothelial cell protein was identified as myosin by the characteristic ATPase profile, that is, the ATPase was activated by Ca2 + and EDTA and inhibited by Mg2 +. On sodium dodecyl sulphate polyacrylamide gel electrophoresis, the endothelial cell myosin heavy chain migrated with a molecular weight of 200 000 as did rabbit uterine and human platelet myosin heavy chains. A crude preparation of the endothelial cell myosin reacted immunologically with an antiserum to platelet myosin, a smooth muscle type of myosin. In indirect immunofluorescence studies, antiserum to the purified endothelial cell myosin stained cultured endothelial cells in a fibrillar pattern. The fibrillar pattern was more intense when the endothelial cells were stained with antiserum to platelet myosin. The presence of myosin in the endothelial cell provides a basis for the contractility of these cells. This contractile property may plan an important role in the physiologic function of these cells.


Amino acid sequence studies on the alpha chain of human fibrinogen. Covalent structure of the alpha-chain portion of fragment D. The alpha-chain portion of fragment D has been purified from an exhaustive plasmic digest of human fibrinogen. The major polypeptide species has 91 amino acid residues, although a small amount of a 97-residue chain representing an earlier digestion stage remains. The amino acid sequence of the first 44 residues was determined by stepwise degradation with an automatic solid-phase sequencer. Another large stretch of sequence was revealed by the finding that the alpha chain of fragment D overlaps the cyanogen bromide fragments alphaCNIVA and alphaCNIII (Doolittle, R. F. Cassman, K. G., Cottrell, B. A., Friezner, S. J. Hucko, J. T., and Takagi, T. (1977), Biochemistry 16 (preceding paper in this issue)). The automatic sequencer results were confirmed and extended by the isolation and characterization of 18 of 19 expected tryptic peptides from the fragment D alpha chain. As a result, almost the entire sequence has been obtained. The overlap with key cyanogen bromide fragments has also allowed us to propose an order for the first 198 residues of the fibrinogen alpha chain. A striking homology with the gamma chain and beta chain is apparent which has interesting structural implications.


Filamin, a new high-molecular-weight protein found in smooth muscle and nonmuscle cells. Purification and properties of chicken gizzard filamin. Filamin, a major high-molecular-weight protein of chicken gizzard smooth muscle, was purified to homogeneity by salt extraction, ammonium sulfate precipitation, agarose gel filtration, and diethylaminoethylcellulose ion-exchange chromatography. Purified filamin is an asymmetric oligomer consisting of two large subunits of identical size (2 X 250 000 daltons) as indicated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, chemical cross-linking, sedimentation analysis (s10, wo = 10S) and Stokes'radius estimation (a = 120 A), It has no intersubunit disulfide but appears from oxidation studies to have adjacent thiols near the subunit interface. Filamin contains no amino sugars, methylated lysine, methylated histidine, or hydroxyproline, nor does it exhibit myosin-like ATPase activities. Its amino acid composition and physical properties differ from those of gizzard myosin, for which a pruification procedure is described. Filamin and the protein spectrin of erythrocyte membranes have strikingly similar physical properties, but they are chemically distinct.


Evidence supporting the identity of beef heart mitochondrial chloroform-released adenosine triphosphatase (ATPase) with coupling factor I. Highly purified mitochondrial chloroform-released beef heart ATPase had molecular weight 330 000, five bands (alpha, beta, gamma, delta, epsilon) in sodium dodecyl sulfate gel electrophoresis and could restore the oxidative-phosphorylation function of A particles. Maximal inhibition (90%) of the enzyme by N,N'-dicyclohexylcarbodiimide was achieved at a molar ratio of inhibitor to protein of 30 : 1. Chloroform introduced into an aqueous solution of beef heart coupling factor I protected it from cold inactivation.


Specific photolabelling of beef-heart mitochondrial ATPase by 8-azido-ATP. 1. 8-Azido-ATP is a suitable photoaffinity label for beef-heart mitochondrial ATPase (F1) 2. 8-Azido-ATP is hydrolysed slowly by F1 in the dark. Photolysis at 350 nm in the presence of F1 leads to inhibition of the ATPase activity. The presence of ATP during illumination prevents the inhibition. Illumination of F1 in the absence of 8-azido-ATP causes no inhibition. 3. Added Mg2+ is not necessary for the binding of the 8-azido-ATP to F1. 4. 8-Azido-ATP binds specifically to the beta subunits of F1. 5. The ATPase activity is completely inhibited when 2 mol of 8-azido-ATP are bound per mol F1.


A study of the dynamic properties of actomyosin systems by quasi-elastic light scattering. A laser light source and a digital autocorrelator were employed in the study of the molecular dyanmics of acto-heavy meromyosin during the splitting of ATP. Low protein concentrations were used, so that molecular and not gel properties were evident. The addition of Mg2+ to acto-heavy meromyosin solutions in the presence of ATP caused a marked widening of the spectrum at high scattering angles. No such change was observed when chemically inactivated heavy meromyosin was used when actin was cross-linked or when the proteins were in a high ionic strength solution. The data can be interpreted in terms of pronounced change in flexibility of acto-heavy meromyosin induced by active mechanochemical coupling.


Isolation and characterization of the surface membranes of fast and slow mammalian skeletal muscle. Fast (extensor digitorum longus) and slow (soleus) rat skeletal muscles served as the source for isolation and biochemical comparison of two distinct surface membrane fractions with properties of the sarcolemma and transverse tubular system. Enriched sarcolemmal membrane from soleus demonstrated a lighter density after sucrose density centrifugation. Sialic acid content was 1.5-fold higher in soleus (62 nmol/mg) than extensor (40 nmol/mg). The specific activity of (Na+ + K+ + Mg2+)-ATPase was similar (1.40 and 1.65 micronmol Pi/mg per 5 min) with the soleus enzyme displaying a (1) greater resistance to inhibition by ouabain, and (2) broader ionic ratio (Na+/K+) requirement than extensor enzyme. The polypeptide and phospholipid composition showed no major differences between the two muscle types. The second surface membrane fraction, tentatively identified as transverse tubule, differed in membrane composition. The major polypeptide of extensor was of 95 000 molecular weight whereas for soleus a Mr=28 000 species was dominant. Total phospholipid content of soleus was 1.5-fold greater than extensor due mostly to increased levels of phosphatidylcholine and phosphatidylethanolamine. Endogenous membrane protein kinase for the 28 000 molecular weight polypeptide was found exclusively in this membrane. The reaction conditions were identical for extensor and soleus since both required divalent cations (Ca2+ and Mg2+) and neither was affected by cyclic AMP. Soleus showed a 2-fold higher capacity for phosphate incorporation than extensor. These studies show that surface membrane fractions derived from fast and slow muscles differ in terms of functional and compositional properties. These differences are specific not only for the surface membrane but for the muscle type and may relate to the known physiological differences observed between fast and slow mammalian muscle.


The effect of calcium ion transport ATPase upon the passive calcium ion permeability of phospholipid vesicles. The uptake and release of Ca2+ by sarcoplasmic reticulum fragments and reconstituted ATPase vesicles was measured by a stopped-flow fluorescence method using chlortetracycline as Ca2+ indicator. Incorporation of the Ca2+ transport ATPase into phospholipid bilayers of widely different fatty acid composition increases their passive permeability to Ca2+ by several orders of magnitude. Therefore in addition to participating in active Ca2+ transport, the (Mg2+ + Ca2+)-activated ATPase also forms hydrophilic channels across the membrane. The relative insensitivity of the permeability effect of ATPase to changes in the fatty acid composition of the membrane is in accord with the suggestion that the Ca2+ channels arise by protein-protein interaction between four ATPase molecules. The reversible formation of these channels may have physiological significance in the rapid Ca2+ release from the sarcoplasmic reticulum during activation of muscle.


Purification and characterization of (Na+ + K+)-ATPase. VI. Differential tryptic modification of catalytic functions of the purified enzyme in presence of NaCl and KCl. 1. Two distinct patterns of tryptic modification of the catalytic functions of purified (Na+ + K+)-ATPase can be related to the two previously described patterns of enzyme inactivation and cleavage of the large chain seen with NaCl and KCl (Jorgensen, P.L. (1975) Biochim. Biophys. Acta 401, 399-415). 2. With NaCl, in phase A, the rapid inactivation of 50-55% of the (Na+ + K+)-ATPase activity is associated with loss of 85% of the K+-phosphatase activity and an increase in Na+-ADP-ATP exchange activity to 150% of control. ATP binding and phosphorylation are unchanged and the inactivation may result from cleavage of bonds within the large chain which are involved in dephosphorylation reactions. In phase B with NaCl, ATP binding and phosphorylation are lost slowly in parallel to inactivation of (Na+ + K+)-ATPase and cleavage of the large chain to a fragment with Mr=78 000. 3. With KCl, cleavage of the large chain to almost equal fragments abolish ATP binding and phosphorylation in parallel to the inactivation of (Na+ + K+)-ATPase. An additional split seems required for inactivation of the K+-pNPPase activity. 4. After completion of the digestion in phase A with NaCl a stable preparation can be isolated in which the activity of (Na+ + K+)-ATPase is 40%. ATP binding and phosphorylation are 90%, K+-phosphatase is 15%, and Na+-ADP-ATP exchange is 150% of control. We currently examine if these levels are related to changes in phosphorylation kinetics. 5. The ATP binding area is much more stable to trypsin with NaCl than with KCl, but loss of the binding capacity is in both cases correlated to a distinct cleavage of the large chain. The relationship between the fractional loss of ATP binding and cleavage of the large chain suggests that the nucleotide binding area is confined to one of the two large chains in the protein complex with Mr=270 000 which binds one molecule of ATP. 6. The data also suggest that the phosphatase site is remote from the ATP binding area. It is proposed that the protein complex with Mr=270 000 contains two large chains with different catalytic functions and that each chain forms a cation channel.


Is there a plasma membrane-located anion-sensitive ATPase? A study of the intracellular localization of HCO-3-stimulated, SCN--inhibited magnesium-dependent ATPase was performed in gill tissue of the rainbow trout (Salmo irideus), rabbit kidney and rabbit gastric mucosa. Tissue homogenates were subjected to centrifugal fractionation, and the microsomal (60 min 100 000 X g) and light mitochondrial (20 min 20 000 X g) fractions were further fractionated by density gradient centrifugation. Subfractions were characterized by marker enzyme assays and electron microscopic observation. In trout gill indications for an exclusively mitochondrial localization were found. In kidney no definite conclusions could be drawn. In rabbit gastric mucosa initially an apparently non-mitochondrial HCO-3-stimulated ATPase, in addition to a mitochondrial one, was found and its characteristics were studied. Further studies showed that this ATPase also appears to be of mitochondrial origin and probably represents mitochondrial inner membranes. Possible explanations for earlier conflicting reports concerning the localization of this enzyme in gastric mucosa and other tissues are discussed.


Crossed immunoelectrophoresis of bovine milk fat globule membrane protein solubilized with non-ionic detergent. Detergent solubilized bovine milk fat globule membrane material studied by crossed immunoelectrophoresis combined with histochemical techniques revealed four major protein complexes. All four were found to bind to concanavalin A and three were identified as sialoglycoproteins. Xanthine oxidase activity was associated with the non-sialoglycoprotein precipitate. Immunoabsorption with intact milk fat globules showed an internal location of the xanthine oxidase, whereas the three other main proteins plus Mg2+-ATPase and 5'-nucleotidase were disposed on the outer membrane surface. The major proteins from milk fat globule membrane and membrane material isolated from skim milk showed immunochemical identity.


Seasonal variation in the active transporting ability and in the membrane ATPase activity of the frog intestinal epithelium. The active sugar and amino acid transport in the small intestine of the American leopard frog (Rana pipiens) and a species of European frog (Rana esculenta) decreases during the winter months. Parallel with this the (Na+, K+)-stimulated ("pump") ATPase activity is markedly depressed. No seasonal changes are observed in the intestine of the tropical bullfrog (Rana catesbeiana). It is assumed that the low pump-ATPase activity is caused by the hibernation of the frogs living in moderate or subtropical areas and is connected to a biological clock. The decreased active transport of non-electrolytes appears to be a consequence of the change of the ATPase activity.


Interaction of caffeine with the DNA of Chinese hamster cells. Incubation of Chinese hamster cells with labelled caffeine leads to transfer of radioactivity to DNA. This association occurs during the S phase of the cell cycle and involves parental as well as newly synthesised strands. The replacement of thymidine by BrdUrd prevents the incorporation radioactivity from caffeine into the DNA strands containing BrdUrd. Thymine is the only base which becomes labelled and data suggesting the participation of methyl groups of caffeine in the biosynthesis of thymine are presented. Ultraviolet irradiation increases the incorporation of radioactivity from caffeine to DNA.


A complete cleavage map of Neurospora crassa mtDNA obtained with endonucleases Eco RI and Bam HI. A physical map of Neurospora crassa mitochondrial DNA has been constructed using specific fragments obtained with restriction endonucleases. The DNA has 5 cleavage sites for endonuclease Bam HI, 12 for endonuclease Eco RI and more than 30 for endonuclease Hind III. The sequence of the Eco RI and Bam HI fragments has been established by analysis of partial fragments. By digestion of the Eco RI fragments with Bam HI, a complete overlapping map has been constructed. The position of the largest Hind III fragment on this map has also been determined. The map is circular and the added molecular weight of the fragments is 40 - 10(6), which is in good agreement with earlier measurements on intact DNA, using the electron microscope.


A comparative study on the influence of cysteamine and metyrapone on mixed-function oxygenase activities in variously pretreated liver microsomes from rats and mice. It has been found that metyrapone can inhibit both type I and type II mixed-function oxygenase reactions, while cysteamine inhibits only type I activity in this mammalian system. Following pretreatment with phenobarbital and 3-methylcholanthrene the half-maximal inhibiting concentrations for the O-demethylation of paranitranisol are increased for cysteamine and decreased for metyrapone. Both cysteamine and metyrapone give type II binding spectra with oxidized cytochrome P-450. The negative and positive peaks are at 393 and 426 nm respectively for metyrapone, and 410 and 434 nm for cysteamine. Cysteamine showed no binding comparable to that of metyrapone for reduced cytochrome P-450. Metyrapone showed little or no inhibition of the NADH cytochrome-c reductase (EC 1.6.1.1) or NADPH (EC 1.6.2.3) cytochrome-c reductase while cysteamine had a more or less strong inhibiting effect depending on the pretreatment of animals. Neither the binding to P-450 heme nor the inhibition of NADH and NADPH cytochrome-c reductase correlates well with cysteamine inhibition of total activity. It is therefore suggested that cysteamine reacts with an intermediate electron carrier of non-heme iron or glycoprotein character thus inhibiting mixed-function oxygenase activity.


Thymine 7-hydroxylase from Neurospora crassa. Substrate specificity studies. A partially purified preparation of thymine 7-hydroxylase (thymine, 2-oxoglutarate : oxygen oxidoreductase (7-hydroxylating), EC 1.14.11.6) from Neurospora crassa was incubated with a number of pyrimidines chemically related to tyymine. 1. Pyrimidines with oxygen or sulfur substituents on atoms Nos. 2 and 4 as well as an alkyl group on atom Nos. 1 or 5 were substrates. 2. Km values were determined for 1-methyluracil, 1-ethyluracil, thymine, 6-azathymine, 1-methylthymine, 1-ethylthymine, 5-formyluracil and 5-hydroxymethyluracil. 3. Uracil was identified as one of the metabolites after incubation with 1-methyluracil. The one-carbon metabolite has not been  characterized. 4. Several pyrimidines with polar groups on atoms Nos. 2 and 4 were inhibitory. 5. Addition of 1-methyluracil, 1-methylthymine, 1-ethylthymine or 5-hydroxymethyluracil to incubations with thymine and 2-oxo[1-14C1]glutarate did not result in additional formation of 14CO2, indicating that the same enzyme acts on the different compounds. It has previously been found (Bankel, L., Holme, E., Lindstedt, G. and Lindstedt, S. (1972) FEBS Lett. 21, 135-138) that a mutant strain of N. crassa which is devoid of thymine 7-hydroxylase activity also lacks ability to perform the coupled oxygenation of 2-oxoglutarate and 1-methyluracil, 5-hydroxymethyluracil and 5-formyluracil, respectively. It is concluded that one and the same oxygenase is responsible for the activities studied.


Kinetic studies of the urokinase-catalysed conversion of NH2-terminal glutamic acid plasminogen to plasmin. Initial velocities for the urokinase (EC 3.4.99.26)-catalysed conversion of glutamic acid plasminogen to plasmin (EC 3.4.21.7) have been determined at various urokinase and glutamic acid plasminogen concentrations. As has been found for the corresponding reaction with lysine plasminogen this conversion obeys the Michaelis rate equation. The apparent Michaelis constants are of the same order of magnitude for lysine and glutamic acid plasminogens. The difference in conversion rates for the reactions has been shown to be connected with their having different catalytic constants. The data were analysed according to two reaction schemes, in one of which only one peptide bond is split during the glutamic acid plasminogen-plasmin conversion and in the other of which the cleavage of two peptide bonds with the obligatory formation of an intermediate plasminogen is assumed. The results favour the former.


Kinetics studies on the interaction between ouabain and (Na+,K+)-ATPase. The association and dissociation rate constants for the interaction of [3H]-ouabain with partially purified rat brain (Na+,K+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) in vitro were estimated from the time course of the [3H]-ouabain binding observed in the presence of Na+, Mg2+ and ATP by a polynomial approximation-curve-fitting technique. The reduction of the association rate constant by K+ was greater than its reduction of the dissociation rate constant. Thus, the affinity of Na+,K+)-ATPase for ouabain was reduced by K+. The binding-site concentration was unaffected by K+. Consistent with these findings, the addition of KCl to an incubation mixture at the time when [3H]-ouabain binding to (Na+,K+)ATPase is close to equilibrium, caused an immediate decrease in bound ouabain concentration, apparently shifting towards a new, lower equilibrium concentration. Dissociation rate constants which were estimated following the termination of the ouabain-binding reaction were different from those estimated with above methods and may not be useful in predicting the ligand effects on equilibrium of the ouabain-enzyme interaction.


Studies on the initial phase of dynein ATPase activity. Kinetic measurement of the reaction of dynein ATPase (ATP phosphohydrolase, EC 3.6.1.3) extracted from the gills of Mytilus edulis shows that in the presence of Mg2+ there is a very rapid initial liberation of Pi from the dynein-ATP system, followed by a slower liberation in the steady state. In view of following results, we have confirmed that this phenomenon is not due to the accumulation of end products, a fall in substrate concentration, nor to the presence of labile impurities in ATP but is due to the catalytic activity of dynein ATPase. 1. The replacement of native dynein by heat denatured dynein or other kinds of Mg2+-ATPase could not produce such a burst phenomenon under the same condition. 2. Both the rate of initial burst and that of steady state were proportional to enzyme content over a wide range under our standard condition. 3. Initial burst was also observed under the constant ATP level by using a ATP generate system. 4. Preincubation of dynein with Pi prior to initiation of the reaction did not eliminate the initial burst. Some properties of the initial rapid liberation of dynein ATPase were also examined. These are shown below. 5. The free ADP liberation did not show any initial burst though the Pi liberation did in the initial phase and the rate of free ADP liberation was almost equal to that of Pi liberation of the steady state. 6. Mg2+ was more effective than Ca2+ for the appearance of the initial burst while the liberation of Pi in the steady state was activated more by Ca2+ than by Mg2+. The addition of K+ in the presence of Mg2+ resulted in a marked increase of Pi liberation in the steady state but not in the initial state. 7. The activation energy of the initial burst was 9.7 kcal, which is slightly smaller than that of myosin ATPase.


The effect of chelators on Mg2+, Na+-dependent phosphorylation of (Na+ + K+)-activated ATPase. 1. The effect of free Mg2+, MgEDTA and MgCDTA on the phofphorylation of the (Na+ + K+)-activated ATPase (ATP phosphohydrolase, EC 3.6.1.3) has been studied. 2. 10 mM trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) added simultaneously with [gamma-32P]ATP to a solution containing the enzyme, 1 mM Mg2+ and 150 mM Na+ does not prevent formation of phospho-enzyme. When [gamma-32P]ATP is added after CDTA the level of phospho-enzyme obtained decreases with increase in the time interval between addition of CDTA and ATP. The inability of CDTA to prevent the formation of phospho-enzyme becomes more pronounced when the medium contains MgEDTA. In the presence of CDTA the maximum amount of phospho-enzyme formed increases with the MgEDTA concentration. 3. Without CDTA the steady-state level of phospho-enzyme is directly proportional to the logarithm of free Mg2+ concentration. Neither with suboptimal nor with optimal concentrations of free Mg2+ does MgEDTA have an effect on the level of phospho-enzyme formed. 4. Using the phospho-enzyme level as a measure of free Mg2+ the experiments show that CDTA reacts slower with Mg2+ than does EDTA, but the stability constant of MgCDTA complex is higher than of MgCDTA, complex. 5. Due to the higher stability constant, of MgCDTA, as compared to MgEDTA, addition of CDTA to a medium containing free Mg2+ and MgEDTA will not only chelate the free Mg2+, but it will also shift the equilibrium from MgEDTA towards MgCDTA, i.e. MgEDTA acts as a source of free Mg2+ which is then chelated by CDTA. The experiments show that it takes minutes before Mg2+, EDTA and CDTA come to equilibrium. Provided the dissociation of MgEDTA is faster than the formation of the MgCDTA complex, the medium will contain a concentration of free Mg2+ which at any given instant is near in equilibrium with a slowly decreasing concentration of MgEDTA; this free Mg2+ can support phosphorylation. This can explain why the rate with which CDTA stops phosphorylation decreases with an increase in the MgEDTA concentration. 6. When phosphorylation is stopped by addition of unlabelled ATP, the rate of dephosphorylation is faster than when it is stopped by addition of CDTA both with and without EDTA in the medium. CDTA reacts too slowly with Mg2+ to be used as a chelator in studies where a fast removal of Mg2+ is required. 7. A previous finding has been verified, namely that the rate of spontaneous, of K+-stimulated and of ADP-stimulated dephosphorylation is independent of the Mg2+ concentration during formation of phospho-enzyme.


Functional integrity of the SH1 site in myosin from hypertrophied myocardium. The hypothesis that an alteration in the SH1 site of hypertrophy myosin is reponsible for the reduced Ca2+-stimulated ATPase activity is examined.The functional integrity of the SH1 site was evaluated by measurement of the (K+)-EDTA-stimulated and Mg2+-inhibited ATPase activities. Neither activity differed from control although the Ca2+-stimulated ATPase of the same preparations was significantly reduced. The reduction in Ca2+-activated ATPase was independent of ionic strength. Titration with N-ethylmaleimide elevated the Ca2+-stimulated ATPase of hypertrophy myosin to the same peak activity as control. Actin-stimulated ATPase activity of hypertrophy myosin was also reduced. The results indicate that the SH1 of hypertrophy myosin is functionally intact for (K+)EDTA-stimulated ATPase and Mg2+ inhibition, but functionally deficient with regard to Ca2+-stimulated and actin-activated ATPase activities. This implies a partition of the functional aspects of SH1.


Correlation between microsomal (Na+ + K+)-ATPase activity and [3H]ouabain binding to heart tissue homogenates. ATP plus Mg2+ plus Na+ supported [3H]ouabain binding to canine left ventricular tissue homogenates and microsomal (Na+ + K+)-ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity from the same tissue were measured. A linear relationship was found between the initial velocity of [3H]ouabain binding to tissue homogenates and microsomal (Na+ + K+)-ATPase activity from the same tissue in the presence and absence of in vivo bound digoxin. In vivo bound digoxin reduced both measurements. With tissue from digoxin-free hearts, a linear relationship was also obtained between the initial velocity and the maximum level of [3H]ouabain binding to tissue homogenate. Binding of [3H]ouabain to whole tissue homogenate is a convenient method for estimating (Na+ + K+)-ATPase activity in small left ventricular biopsy samples.


The pre-steady-state hydrolysis of ATP by porcine brain (Na+ + K+)-dependent ATPase. The hydrolysis of [gamma-32P]ATP by porcine brain (Na+ + K+)-stimulated ATP phosphohydrolase (EC 3.6.1.3) has been studied at 28 degree C in a rapid mixing quenched-flow apparatus. An "early burst" in the release of Pi from ATP has been observed when the enzyme is mixed with ATP, Na+ and a relatively high concentration of K+ (10 mM) but the burst is less pronounced with 0.5 mM K+. This "early burst" of Pi release is suppressed when the enzyme is pre-mixed with 10 mM K+ or 20% (v/v) dimethylsulphoxide before mixing with ATP and Na+, and premixing of enzyme with Na+ antagonizes this effect of dimethylsulphoxide. The results have been analysed by a non-linear least squares regression treatment and are consistent with a mechanism involving three steps, one of which may be a relatively slow change in enzyme conformation following release of Pi from its covalent linkage with the enzyme, in addition to formation of the enzyme-substrate complex. Rate constants (and S.E.) for these steps have been calculated and the roles of phospho-enzyme and other intermediates in the reaction mechanism of the transport ATPase are dicussed.


Lipoprotein lipase in cultured heart cells: characteristics and cellular location. Lipase activity extracted from cultured neonatal rat heart cells was characterized and identified as lipoprotein lipase. Enzyme activity was stimulated by human apoC-II and rat serum; serum stimulation was prevented by human apoC-I and by apoC-II. Lipolysis was maximal at pH 8.0 and was inhibited by protamine sulfate, NaCl, and high concentrations of heparin. About 50% of heart cell lipase activity applied to heparin-Sepharose bound to the gel and was eluted with a NaCl gradient. A peak of lipase activity was observed at 0.84 M NaCl. Neonatal rat heart cells in culture are a mixture of muscle and non-muscle cells. To determine the cellular location of the lipoprotein lipase, enzyme activity and muscle cell content of the cultures were determined. Myosin ATPase was used as an index of muscle cell content since ATPase specific activity correlated (r = +0.97) with muscle cell content determined immunofluorescently. When muscle cell content of cultures was decreased or increased by differential plating, lipase specific activity was constant. Moreover, lipase specific activity was constant during culture growth despite a decrease in muscle cell content. It was concluded that lipoprotein lipase activity of cultured heart cells is not associated solely with either muscle or non-muslce cells.


Renal calcium binding protein in the diabetic and vitamin D-depleted rat. A calcium binding protein has been purified 220 fold from rat kidney. The molecular weight of this protein (26 000-28 000) is more than double that of the duodenal calcium binding protein of the rat. In response to the stimuli of both streptozotocin diabetes and depletion and repletion with vitamin D, changes in the renal protein are minimal. This contrasts markedly with responses of the duodenal protein to the same stimuli: (a) there was marked depression of duodenal calcium binding protein by vitamin D depletion and diabetes; (b) duodenal calcium binding protein was restored by vitamin D treatment of depleted rats. The renal protein appears to be identical with a previously described 28 000 molecular weight protein from the kidney purified by a different technique (Hermsdorf, C.L. and Bronner, F. (1975) Biochim. Biophys. Acta 379, 553-561). In contrast to findings of the current study, previous investigators were unable to isolate the protein from vitamin D-deficient rats and postulated vitamin D dependence. The protein activator of cyclic AMP phosphodiesterase is a calcium binding protein found in many tissues including kidney. Based on lack of response to stimuli we used and similarity in method of isolation and properties, our renal calcium binding protein may be this protein activator.


Production of enterochelin by Escherichia coli 0111. The major neutral iron-transporting compound produced by Escherichia coli 0111/K58/H2 has been isolated from iron-deficient cultures of the organism and compared with the corresponding compound, enterochelin, produced by E. coli K12. The product contained serine and 2,3-dihydroxybenzoic acid and formed a complex with Fe3+. Since the PMR spectra of the products from the two strains were identical, it was concluded that E. coli 0111 also secreted enterochelin under iron-deficient conditions. Although it was not possible to establish the optical configuration of the serine residues in the molecule, the CD spectra of the metal free and Fe3+,  complexes were found to be of the same sign and magnitude. The spectra show that metal binding results in considerable conformational changes in the enterochelin molecule. The biological properties of the two compounds appear to be identical as judged by their ability to abolish the bacteriostatic effect of serum on E. coli 0111.


[Some peculiarities of the reaction kinetics of ATP hydrolysis by Ca2+-dependent ATPase of chloroplasts]. The dependence of the reaction rate of ATP hydrolysis by CF1-ATPase of chloroplasts on the substrate (CaATP) concentration is of complex nature. It is assumed that such dependence may be due to either conformational changes of the enzyme during the increase in CaATP content in the reaction medium, or consecutive binding of the substrate to its active sites, differing in their affinity to the substrate of the reaction.


[Study of ADP effect on brain Na+, K+-ATPase]. A mechanism of K-insensitive, ouabain-dependent liberation of Na+ from the cell during an increase in ADP intracellular concentration is studied. It is shown that the increase in the ADP/ATP ratio does not change the Na+, K+-ATPase affinity to K+ ions and does not result in the Na-activated, K-independent ATPase reaction. ADP protects ATPase from the inhibition by ouabain which is accounted for by a decrease in the concentration of a glycoside-sensitive form of the enzyme E2-P due to a turnover of the phosphokinase step of the reaction, but not due to the binding of free Mg2+ ions. The results obtained suggest that the increase in ADP concentration within the cell activates Na-Nan exchange along Na-transporting channels of the ionic pump.


[Isolation of highly active preparations of sarcoplasmic reticulum and Ca2-dependent ATPase from cardiac muscle]. A microsomal preparation with a high ability for Ca2+ uptake has been isolated from pigeon heart. A method of further purification of Ca2+-accumulating system of heart, based on the ability of sarcoplasmic reticulum for the energy-dependent Ca2+ accumulation in the presence of oxalate, has been developed. Upon centrifugation in the gradient of sucrose and KCl concentration the fragments of sarcoplasmic reticulum, rendered "heavy" by calcium oxalate, can be separated from foreign cell membranes. The main component of heart "calcium pump" is Ca2+-dependent ATPase (making up to about 50% of all proteins of the purified reticulum), having a molecular weight of 100.000--105.000. Specific activity of heart Ca2+-ATPase as well as the ability of purified heart sarcoplasmic reticulum for Ca2+ uptake are only slightly less than those of the skeletal muscle reticulum. The data obtained suggest that heart sarcoplasmic reticulum may be efficient for providing heart muscle relaxation.


[Mechanism of oxidation reaction of NADH models and phynylglyoxal with hydrogen peroxide. Hypothesis on separate transport of hydrogen and electron atom in certain enzymatic reactions with the participation of NADH and NADPH]. Kinetics of co-oxidation of 1-benzen-3-carbamido-1,4-dihydropyridine (BDN) and phenylglyoxal (PG) with hydrogen peroxide is studied. Dimeric product (di-e11-benzen-5-carbamido-1,2-dihydropyridyl-2]) is found to be formed at pH 9, and quaternal pyridinium salt (BNA)--at pH 7. Molecular oxigen is determined to participate in the reaction at pH 7. Copper (II) ions catalyze this process. Significant catalytic effect of p-dinitrobenzen (p-DNB) is found. The reaction mechanism is postulated to form hydroperoxide from PG and hydrogen peroxide which are capable to split the hydrogen attom from dihydropyridine, molecular oxigen or p-DNB being an acceptor of the electrone. Hypothesis on separate transfer of hydrogen atom and electrone in biological systems are proposed.


[Study of the interaction of Na+ and K+-ATPase of erythrocytes with ouabain. Effect of acetyl phosphate and p-nitrophenyl phosphate]. Effects of ATP, acetyl phosphate (AcP) and p-nitrophenyl phosphate (p-NPP) on the inhibition of the Na+, K+-ATPase activity were studied. ATP, AcP and p-NPP were found to facilitate the ouabain-induced inhibition of the enzyme activity only after the injection of these phosphorylyzing agents into the erythrocyte ghosts. Inside the ghosts Na+ ions enhanced the effects of the phosphorylyzing agents. K+ ions in the environment removed the stimulating effects of ATP, AcP and p-NPP on the ouabain-induced inhibition of Na+, K+-ATPase activity. It is concluded that the sites of AcP and p-NPP hydrolysis as well as the active center for ATP are localized on the inner surface of the cell membrane.


[Determination of the secondary prothrombin structure under different degrees of citraconylation by means of ring dichroism]. A process of spontaneous activation of bovine prothrombin under acylation of cytacone anhydride is studied by means of ring dichroism and disc electrophoresis in sodium dodecylsulphate. Ring dichroism data have shown that native prothrombin contains 14% of alpha-helical structures, 36% of beta-structures and 50% of random coil. Cytraconylation results in a fragmentation of native prothrombin chain and is accompanies by the decrease of alpha-helical regions in the fragments formed. Poly-L-lysine, modified by citracone anydride, does not form alpha-helical regions.


[Changes of prothrombin activity by plasmin immobilized on L-lysine Sepharose 4B]. It is demonstrated that the method of immobilization of human plasmin by its adsorption on L-lysine-Sepharose 4B can be successfully applied for bovine plasmin isolation. Proteolysis of bovine prothrombin by bovine prothrombin by bovine plasmin preparations of high caseinolytic activity results in the increase of prothrombin level. Prothrombin proteolysis by bovine plasmin preparations of high caseinolytic activity results in the decrease of prothrombin level.


[Changes in the ionization equilibrium of erythrocyte suspension under heating]. The effects of ionic strength, urea, calcium and fluorine ions, ouabain and cholinesterase inhibitors on the changes in the ionization equilibrium of an erythrocyte suspension under heating were studied. Proton release by erythrocytes was compared to a release of potassium ions and hemoglobin from the cells. The proton release under heating is mainly determined by the physico--chemical properties of superficial structures of erythrocytes and does not depend on the activity of cholinesterase, ATPase and glycolytic processes.


[Resting membrane potentials and miniature postsynaptic potentials of frog intrafusal muscle fibers]. Electrophysiological charact eristics of frog intrafusal muscle fibers were studied by means of the intracellular potential recording. Analysis of distribution of the amplitude of miniature postsynaptic potentials suggests the nonuniformity of the intrafusal muscle fibers. A conclusion was drawn on the presence of three types of intrafasal muscle fibers in frog muscle spindle: single-innervated, multiple innervated, and fibers with both types of innervation.


[Effect of tetanus toxin and colchicine on synaptic membranes of rat cerebral cortex]. It was shown that purified tetanus toxin did not influence the activity of the Na, K-ATP-ase fractions of the synaptic membranes of the rat cerebral cortex, it had no effect on the inhibition of Na, K,-ATP-ase under electrical stimulation of the synaptic membrane suspension, or the GABA--3H binding by the synaptosomes in vitro. Tetanus toxin (400--4000 DLM) and colchicine (1 mM) induced a decrease of osmotic sensitivity of the nerve endings. Colchicine in low concentrations (10(-5)-10(-3) M) failed to influence Mg-and Na, K-ATP-ases, but considerably inhibited both ATP-ases at higher concentrations.


[Effect of amizil and arecoline on the activity of Na, K-ATP-ase and concentration of Na+ and K+ ions in rat brain]. A study was made of the activity of Na, K-ATP-ase and the Na+ and K+ content in the brain of rats with the action of arecoline and amizyl. Both arecoline and amizyl increased the Na, K-ATP-ase activity. This could be associated with the changes in the redistribution of the Na+ and K+ ions in the nerve cell. Arecoline proved to cause changes in the electrolyte distribution by the depolarization type, whereas amizyl--by the type of hyperpolarization of the nerve cell membrane.


[Monoamine oxidase type B and the function of Ca2+, Mg2+-dependent adenosine triphosphatase of preparations of sarcoplasmic reticulum vesicles]. Oxidative deamination by monoamine oxidases of B type in the preparations of sarcoplasmic reticulum vesicles from rabbit skeletal muscles of beta-phenylethylamine or benzylamine was accompanied by a decrease of both the active transport of Ca2+ into the vesicles and Ca2+, Mg2+-dependent ATP-ase activity. This decrease was prevented by pretreatment of the vesicles with deprenyl, a specific monoamine oxidase type B inhibitor. The aldehydes formed in the course of enzymatic deamination of the substrates of type B, monoamine oxidases, are considered as possible participants in the regulation of Ca2+, Mg2+-dependent ATP-ase activity.


[Structure of interphase chromatin in patients with Down's syndrome and their mothers]. A method of fluorescent microscopy with the aid of acridine orange was applied in these studies; some features of the changes in the structure of interphasic chromatin characteristic of their sick children were revealed on the short-term cultures of lymphoyctes obtained from the mothers with children suffering from Down's syndrome. Sibling girls also displayed deviations similar to the changes revealed in their mothers. The data obtained permit to suppose the existence of a definite population of women, peculiarities of whose genotype promoted the appearance on the structural chromatin organization was revealed only in the mothers and sibling girls it is suggested that the mentioned genotype peculiarities were hereditary and connected with genes (or certain chromatin areas) limited by sex.


Leukemic reticuloendotheliosis: "hairy cell leukemia," functional and structural features of the abnormal cell in a patient with profound leukocytosis. The development of profound leukocytosis in a patient with leukemic reticuloendotheliosis (LRE) enabled us to obtain purified LRE cells for the investigation of their structural and functional characteristics. The LRE cells of our patient bore surface immunoglobulin and had complement receptors but did not bear Fc receptors and did not form rosettes with sheep erythrocytes. By electron microscopy, the cells were observed to contain typical ribosome lamella structures and to phagocytize both 0.81 micron latex particles and complement-coated zymosan particles. They were adherent to both glass and nylon wool fibers. The mitogenic response to erythroagglutinating phytohemagglutinin was normal to magnitude but delayed chronologically. The binding of 125I-labeled plant lectins was used to characterize the surface topography of LRE cells. Results of these studies indicated that the LRE cell surface differed significantly from the surface of normal T and B lymphocytes and chronic lymphatic leukemia cells. The LRE cells were capable of both stimulating and responding in a one-way mixed lymphocyte culture. However, the LRE cells were not active as effector cells of either cell-mediated lympholysis, a T cell function, or antibody-dependent cellular cytotoxicity, a null cell function. In contrast, they were effector cells of lectin-induced cellular cytotoxicity showing that they did possess the capacity to function as cytotoxic effector cells. These data indicated that the LRE cells in our patient had surface and functional characteristics of both lymphocytes and monocytes.


Intention myoclonus in Huntington's disease. A patient is described with severe intention myoclonus which was made worse by treatment with L-Dopa and improved by clonazepam. Family history and examination of several siblings suggested the diagnosis of Huntington's disease. Subsequent to improvement of the myoclonus the patient appeared to have the rigid form of Huntington's disease. This case represents a unique expression for an otherwise well defined genetic abnormality. Stimulus activated myoclonus in a common feature of a number of disorders of the nervous system. Intention, or action myoclonus has been the subject of increasing interest because of its association with the syndrome of post-anoxic encephalopathy (1), although it may occur with other disorders as well. Many abnormal movements have been reported in Huntington's disease, but myoclonus is a relatively uncommon feature of this disorder and to our knowledge intention myoclonus has not been reported as a major symptom. We recently have evaluated a patient with disabling intention myoclonus and examined several members of his family who have typical Huntington's disease. We therefore report this case, a unique presentation of an otherwise well described movement disturbance.


Effects of delta9-tetrahydrocannabinol and cannabidiol on a Mg2+-ATPase of synaptic vesicles prepared from rat cerebral cortex. 1. delta9-Tetrahydrocannabinol and cannabidiol both exhibited a concentration-related inhibition of Mg2+-ATPase of vesicles prepared from synaptosomes isolated from rat cerebral cortex. Cannabidiol was about 3 times more potent than tetrahydrocannabinol. 2. These results were similar to those obtained previously using drugs with well established anticonvulsant activity. 3. Tetrahydrocannabinol at a sub-inhibitory concentration (1 micronM) increased the activity of the Mg2+-ATPase relative to values obtained with vehicle controls.


The plasma cholinesterase variants in mentally ill patients. The distribution of the plasma cholinesterase variants found in 1,374 mentally ill patients differs from that of a random control sample. The patients are more likely to have a rare phenotype than an individual from the normal population. None of the diagnostic groups have been shown to differ in the distribution of the E1a gene, but there is strong statistical evidence that Group IV (psychosis) patients have a higher frequency of the E1f gene than the other groups. The overall frequency of the electrophoretic variant C5 + did not differ significantly from that observed in a Caucasian population, with the exception of the increase observed in Group IV c2. Twenty-eight unrelated patients with Huntington's chorea were found to have a significantly altered incidence of the C5 + variant and six patients from this group were found to have the rare E1f gene. Our results indicate that the plasma cholinesterase variants may provide some insight into the inheritance of Huntington's chorea.


The impact of an abnormal child upon the parents. Thirty families with a newborn mongol baby were matched with thirty families with a normal baby. Both groups were followed for eighteen months to two years and interviewed six times. Few differences could be found in the mental or physical health of the two groups of parents, but marital breakdown or severe marital disharmony was found in nine of the mongol families and in none of the controls.


Direct puncture and angiographic evaluation of axillary-to-femoral bypass grafts. Arteriography of patients with axillary-to-femoral bypass grafts can be performed by direct puncture of the graft and passage of a catheter through the graft. This article reviews some technical considerations as well as some potential complications and how they might be minimized. Three cases are presented.


The concept and measurement of bladder work. The methodology for the calculation of bladder work from pressure volume traces has been described. Bladder work values have been calculated from the urodynamic data of 50 male patients. When bladder work values were related to the micturition time and the volume passed, a highly significant correlation was demonstrated with urethral resistance. Bladder work values may be a useful index in the diagnosis of bladder outflow obstruction.


The use of fine double siliconised dacron in urethral replacement. A knitted Dacron mesh was used to form a tube of tissue which was used to bridge at least 5-cm defect in the urethra of 9 dogs. The tube developed a urothelial lining and this showed no change in calibre after 7 months.


Treatment of uncomplicated gonorrhoea with spectinomycin hydrochloride (Trobicin). 110 patients with uncomplicated gonococcal urethritis were treated with 2 g spectinomycin (Trobicin) intramuscularly. Seventy-five patients were available for follow-up and there was only one treatment failure, giving a cure rate of 98-6%. Of the 99 gonococcal isolates, 59 were sensitive to 5 microng/ml of spectinomycin or less and 40 were sensitive to 10 microng/ml. In a small sample of patients 2 g spectinomycin intramuscularly produced a serum concentration averaging 100 microng/ml after one hour.


Penicillinase-producing strains of Neisseria gonorrhoeae isolated in Rotterdam. This appears to be the first report of the isolation of a penicillinase-producing strain of Neisseria gonorrhoeae in the Netherlands. The presence of a penicillinase-producing gonococcal strain (PPGS) was demonstrated in two of a group of three patients with gonorrhoea. In addition, a penicillin- and ampicillin-sensitive gonococcus strain was isolated from all three patients. The PPGS was most probably imported into Holland from the Philippines. The epidemiological and clinical findings and the results of treatment of infection with this PPGS are discussed.