Penetration of an ascitic reticulum cell sarcoma of the golden hampster into the body wall and through the diaphragm.  A scanning electron microscopic (SEM) study.

SEM studies on infiltration of the ascitic form of the hamster reticulum cell sarcoma HaTu 25 into the ventral body wall and through the diaphragm were performed during 6 consecutive days after intraperitoneal transplantation.  The findings allow an interpretation of the course of events based on 3 main stages: 1) Contraction of mesothelial cells with partial exposure of the submesothelial stratum.  2) Preferential attachment of tumor cells to these denuded areas.  3) Advance of tumor cells within defects gradually extening from the submesothelial stratum of the musculature.  These stages were more pronounced and took a more rapid course at the peritoneal side of the diaphragm than at the body wall.  At the pleural side of the diaphragm the appearance of single tumor cells within widened intercellular spaces of the mesothelium was recorded prior to the onset of penetration at the peritoneal surface.  The rapid migration of tumor cells through the diaphragm as well as the particularly intensive tumor infiltration into this organ is thought to be connected with the mechanism of intravasation of tumor cells into the lymphatic plexus of the diaphragm.  During the whole sequence of events, HaTu 25 cells were found to have maintained their spherical configuration and characteristic surface architecture.  Apparently, growth pressure is of minor or no importance in this spacial mode of tumor penetration, rather the action of proteolytic enzymes elaborated by the tumor cells has to be taken into consideration.


[Hormonal therapy of acne vulgaris. Review and results of treatment (author's transl)].

Hormones play a major role in the development of acne vulgaris of pubertal onset. In addition to a seborrhoeic diathesis, an imbalance of oestrogens and androgens, as well as altered enzyme metabolism in the sebaceous glands are factors frequently implicated in the aetiology of this condition. There are many reports on hormonal treatment in the literature; the therapeutic regimes and results achieved varied greatly. The present report is concerned with a discussion of the results obtained in 50 patients with acne vulgaris treated with a drug containing oestriol.


[Changes in the ATPase activity of the brain and erythrocytes in hypoxia].

An effect of hypoxic hypoxia on the total, Mg2+-dependent and Na-K-dependent ATPase activities was studied in rat brain and erythrocytes. In brain tissue under effect of 15 min hypoxia a distinct decrease in all the ATPase activities was observed; the activity of Na-K-dependent ATPase decreased especially distinctly (by 32% of the initial level). Within 90 min of hypoxia the enzymatic activity was partially restored but did not reach the initial level. In erythrocytes 15 min hypoxia was also accompanied by a significant decrease in Na-K-dependent ATPase activity (30.5% of the initial level). But within 90 min the enzymatic activity was distinctly increased (up to 196.4%). Mg2+-dependent ATP-ase was more resistant to oxygen deficiency, Variations in the enzymatic activity were less distinct, although they showed the same pattern as those of the Na-K-dependent ATPase.


[Effect of cytoplasmic and serum factors on the uptake of (3H) thymidine-triphosphate (TTP) by isolated nuclei of liver cells].

The actions of cytoplasm (ultracentrifuged supernatant 105 000 times g) and of serum on the DNA synthesis (uptake of (3H) TTP) by isolated nuclei of liver cells were studied. Cytoplasm from liver cells obtained at various intervals after partial hepatectomy revealed a variable effect only with isolated nuclei of liver cells from partially hepatectomized rats. Cytoplasm obtained from liver cells eight hours after partial hepatectomy failed to show the usual inhibitory effect. Presumably the inhibitory substances normally present in cytoplasm are inactivated within a certain time after partial hepatectomy. Serum of partially hepatectomized animals enhanced the uptake of (3H) TTP by nuclei of normal liver cells and those from partially hepatectomized animals. The stimulating effect was present only in the serum obtained between four and twelve hours after partial hepatectomy. It remains unclear whether the effects described are growth factors specific for the liver.


The effect of 5-hydroxytryptamine and histamine on glycolysis in the mouse brain.

Following the intraventricular injection of 5-hydroxytryptamine into the lateral ventricles of conscious mice, the concentration of brain lactate rose immediately but then decreased significantly compared with the saline injected controls. The concentration of brain glucose increased 20 min after the injection of 5-hydroxytryptamine. These effects of 5-hydroxytryptamine on lactate and glucose were qualitatively similar to those found following the administration of methysergide. After the parenteral administration of 5-hydroxytryptophan, the lactate levels showed a bephasic change but brain glucose was significantly decreased for up to 90 min following the injection. p-Chlorophenylalanine potentiated the effect of 5-hydroxytryptamine by further increasing glucose and decreasing lactate levels. After histamine had been injected into the ventricles, the concentrations of bound glycogen and lactate were decreased whereas free glycogen and glucose were raised. It is suggested that 5-hydroxytryptamine, and possibly histamine, are involved with control of glycolysis in the mouse brain.


Influence of spermine on some membrane-disturbing actions.

The influence of spermine on the potassium loss from yeast cells as a consequence of membrane-disturbing actions was investigated. Spermine interfered strongly in the action membrane-active bactericides on yeast cells; this interference resembles the action of certain metal ions. Spermine provides a protection against the positively charged parasosaniline and accomplishes a strong potentiation of the action of the negatively charged sodium dodecyl sulphate.


Determination of exotoxin in Bacillus thuringiensis cells.

The presence of exotoxin in Bacillus thuringiensis was demonstrated and its quantity in the cells determined. The concentration of exotoxin in the producing microorganism is approximately half the concentration of ATP. Exotoxin is produced at such a rate that the cell excretes 1/5 to 1/4 of its exotoxin content into the medium per minute.


Interactions between black lipid membranes and the loosely bound proteins from erythrocyte membranes.

The loosely bound proteins from human erythrocyte membranes induce an up to more than 103-fold increase in the electrical conductivity of black lipid membranes made from oxidized cholesterol. The conductivity increase continues until the membrane breaks. Its magnitude insreases with increasing protein concentration and salt concentration. The results are compared with earlier results obtained with the strongly bound proteins from erythrocyte membranes.


[Microsome-associated 17 beta-hydroxysteroid dehydrogenases of human placenta, i solubilization, enrichment and separation of two 17 beta-hsd-activities after phospholipase-Treatment].

Treatment of human placenta microsomes with phospholipase A or D inhibits the 17beta-hydroxysteroid dehydrogenase (17beta-HSD) activity, parallel with the hydrolysis of membrane phospholipids. The 17beta-HSD activity of phospholipase treated microsomes is reactivated by synthetic phospholipids. The distribution of 17beta-HSD activity in subfractions of original microsomes and of phospholipase treated microsomes obtained by zonal centrifugation was studied. Solubilization of the microsomal 17beta-HSD was achieved by phospholipase A treatment. Two 17beta-HSD were solubilized from human placenta microsomes by phospholipase A treatment and were further purified by ammonium sulphate precipitation, gel filtration on BioGel A-0.5 m, DEAE-Sephadex chromatography and by isoelectric focusing. The enzymes were purified 25.8 and 17.4 times. The isoelectric points and molecular weights of the two 17beta-HSD were determined. Both enzymes are of a 17beta-HSD type. One of the 17beta-HSD, however, was sensitive to estradiol-17beta, the other to testosterone. The question of whether the two enzymes constitute a monomer and a dimer of the same 17beta-HSD or are completely different enzymes, is discussed.


[Phytochrome system and synthesis of lipoquinones in the plastids of etiolated hordeum seedlings].

Short pulses of red light induce in etiolated barley seedlings an enhanced synthesis of plastidic benzoquinones and vitamin K1, which can be reverted by subsequent irradiation with short pulses of far-red. As compared to the dark there is more formation of plastoquinone-9 than alpha-tocopherol. The enhanced formation of vitamin K1 is coupled with a concomitant decrease in the level of the second naphthoquinone vitamin "K". The data show that active phytochrome Pfr, also in its ground state, induces the light triggered lipoquinone synthesis which is connected to thylakoid formation. The red light induction of enhanced plastoquinone-9 synthesis cannot be fully reverted by subsequent far-red and seems to be a very fast phytochrome response.


[The influence of phytochrome on the formation of individual carotenoids in etiolated hordeum seedlings].

Short time red pulses, given 6 times for 5 min within 36 h, induce in etiolated barley seedlings an enhanced synthesis of the main chloroplast carotenoids beta-carotene, violaxanthine, lutein and neoxanthine. The level of antheraxanthine and zeaxanthine decreases by red light treatment. These red light effects are reverted by subsequent short time far-red pulses. The results show that the white light induced change in the accumulation rate of individual carotenoids is initiated and regulated by active phytochrome Pfr. In the case of neoxanthin and zeaxanthin the red light effects cannot be fully reverted by far-red; this points to a very fast phytochrome reaction.


[The influence of protoplast media on the tumor induction on kalanchoe leaves].

Experiments concerning tumor transformation of plant cells or protoplasts need complete knowledge of the effects of media components. Here inhibiting effects of cellulase Onozuka, glucose, sucrose and mannitol (up to 0.3M), of tissue culture media, and of the antibiotic aureomycin on the tumor initiation process in Kalanchoe leaves are reported. Also the influence of these substances on the growth of the tumor inducing Agrobacterium tumefaciens is studied. The results lead to the design of conditions which should be favourable to in vitro transformation.


[Laws of substrate supply, of cell kinetics and of therapy mechanisms in the intercapillary region of cancer tissue].

Up to now, the microtopography of glucose and O2 concentration in the intercapillary region of tumour tissue has been determined under the simplifying assumption that substrate consumption is constant up to the periphery of the envisaged cylindrical space around a capillary. The general diffusion field equation presented in this paper takes into account that substrate consumption is constant up to the periphery of the envisaged cylindrical space around a capillary. The general diffusion field equation presented in this paper takes into account that substrate consumption decreases in the unsaturated region of cancer cell glycolysis and respiration so that - compared to former computations - the critical supply radii needed for maintained proliferation increase by a factor of almost 1.8. With the aid of its general solution and the discussed parameters, the laws governing substrate supply, cell kinetics and therapy mechanisms in the intercapillary region of intact and treated tumour tissue are presented and discussed. The essential results there are as follows: - Even in case of hyperglycemia it is glucose supply (and not the O2 supply) which determines proliferation and proliferation rate. - Under the conditions of longtime-hyperglycemia (400 mg%), the tumour volume being accessible to the attack of cancerostatica or radiation, is almost three times as high as under standard conditions. - For glucose (and cancerostatica) the time constant for interactions between circulation and tumour tissue near the necrotic region is equal to or greater than 400 min. This is why stimulation of cancer cell glycolysis in the most therapy-resistant cancer cell portions (i. e., increased proliferation rate and tumour hyperacidification) can only be achieved if hyperglycemia is maintained for 24 hours or more. - Therapeutically treated cancer tissue is characterized by the discussed changes in the diffusion field of glucose and O2 as well as a drop in the interaction time constant for glucose.


Non-Gaussian behavior of the EEG in Down's syndrome suggests decreased neuronal connections.

Computer analyses of the electroencephalograms of normal children, and children with Down's syndrome who were mentally retarded, revealed differences in the distribution of amplitude between the two groups. Normal children, in the early postnatal period, generate EEG's which have a non-Gaussian distribution of amplitude that becomes increasingly Gaussian before one year of age and remains so throughout subsequent development. Conversely, the EEG's of children with Down's syndrome exhibit highly non-Gaussian properties at all ages studied. The EEG's of two mentally retarded autistic children did not show this property, so it is not merely a concomitant of mental retardation. The first-order Gaussian distribution may reflect the degree of inteneuronal coupling; since an increased number of connections on any neuron implies decreased functional dependence of that neuron on any other particular single neuron, the present evidence, which indicates stronger interneuronal coupling in mongoloids, suggests that Down's syndrome may be associated with incomplete postnatal development of interconnections between cortical neurons.


Trancutaneous determination of aortic blood-flow velocities in man.

A transcutaneous ultrasonic Doppler technique for measurement of aortic blood-flow velocities has been developed and compared to more established techniques in order to evaluate its potential usefulness.  It is possible by this method to quantitate blood velocity in both the ascending aorta and the aortic arch with ease and reliability.  Ultrasonic access to the aorta from the suprasternal notch proved adequate in more than 90 per cent of the normal subjects examined.  If further clinical trials prove as encouraging, this technique may be of significant value for patient monitoring and cardiac diagnosis.


Cor pulmonale (pulmonary heart disease): present-day status.

In reviewing the present-day status of cor pulmonale, it is clear that considerable progress has been made in almost all instances of the disease.  It is clearly a preventable form of heart disease in most cases and it is treatable and curable in its most common form, i. e., in COPD.  One must agree with Petty, that today we have effective means of caring for the majority of respiratory cripples.  Therapy for lung disease now appears even to have reduced the expected rate of pulmonary function deterioration in some patients.  Surely with improved gas exchange and early detection of respiratory insufficiency the outlook for patients with respiratory diseases leading to cor pulmonale is better today than it was 30 years ago.


Electrocardiographic evidence of left ventricular hypertrophy in otherwise normal children. Clarification by echocardiography.

Eleven apparently normal children whose electrocardiograms demonstrated greatly increased precordial QRS voltages suggestive of left ventricular hypertrophy were studied by echocardiography to determine whether this finding represented an early manifestation of cardiac disease. Echocardiographic measurements of the thickness of the ventricular septum and of the left ventricular posterobasal wall were normal in all 11 subjects when compared with normal values for age. Also within normal limits were left ventricular, left atrial and aortic root internal diameters, left ventricular mass and the motion and structure of the anterior and posterior mitral valve leaflets. We conclude: (1) These children probably have normal hearts; (2) increased precordial QRS voltage in children and adolescents, in the absence of other abnormalities, is an unreliable indicator of left ventricular hypertrophy; and (3) echocardiography is a powerful diagnostic test in determining the significance of abnormal electrocardiograms suggestive of left venticular hypertrophy.


Adenine arabinoside in idoxuridine unresponsive and intolerant herpetic keratitis.

Twenty-two patients with active herpes simplex dendritic keratitis, in whom topical idoxuridine was unsuccessful in controlling their disease, were treated with topical adenine arabinoside (ara-A), a purine analogue effective against many DNA viruses. This drug was an effective antiviral agent in these patients. No signs of ocular or adnexal toxicity were noted.


Harold Cummins--and the birth, growth and development of dermatoglyphics.

The work of Harold Cummins in dermatoglyphics is reviewed, in the areas of populational variability, twin studies, dermatoglyphics of non-human primates, and dermatoglyphic methodology.


Dermatoglyphics of schizophrenics, patients with Down's syndrome and mentally retarded males as compared with Australian Europeans using multivariate statistics.

Dermatoglyphics of schizophrenics, patients with Down's syndrome and mentally retarded males were compared with those of normal Australian Europeans.  A computer programme of multivariate analysis of fifteen dermatoglyphic features was utilized.  This analysis produces two significant variates of each of the populations plotted in two-dimensional space.  The distance, measured in arbitrary units, between any two populations was studied for its significance.  It was noticed that the patients with Down's syndrome separated significantly from the rest of the groups.  The importance of multivariate analysis in the study of dermatoglyphics in comparing two or more populations is discussed.


Changes of sodium transport in erythrocytes of the maturing rabbit.

The red blood cells of New Zealand white rabbits have a low sodium and high potassium content. As the animals mature, the sodium concentration rises and the potassium content falls; studies of red cells from a group of five young and five mature animals revealed a highly significant increase of cell sodium with age that was associated with a significant fall in the rate of ouabain-inhibited active sodium efflux. This difference was still seen when the sodium concentration within the cells from old and young animals was equalized and elevated to saturating levels for active pump efflux. Total sodium efflux, however, increased significantly with age as did total sodium influx so that a steady state was reached. Ouabain-sensitive ATPase activity fell significantly in the cell membranes from older animals and ouabain-insensitive ATPase increased with age. The survival time of 51Cr-labeled red cells was significantly longer in old than in young animals and it is concluded that as the rabbit matures its red cells survive for a longer period and this is associated with the changes of sodium transport and ATPase activity that have been documented.


Action of peptide-B from bovine fibrinogen on ATPase activity and superprecipitation of myosin B.

Bovine peptide-B from fibrinogen was capable of accelerating the structural and enzymatic effects associated with superprecipitation of myosin B. The rate of superprecipitation coupled with the hydrolysis of ATP are increased during the structural transformation. In the concentration range from 10-8 to 10-4 M peptide-B, the rate of superprecipitation is increased 12-fold while the hydrolysis of ATP doubles and the time to reach the final extent of superprecipitation is decreased 68%. Under these same conditions, the hydrolysis of ATP by myosin A was unaffected. The concentrations of magnesium and calcium were between 10 and 20 muM, and no additional divalent metal ions were added to the system. Superprecipitation was treated as a model for muscle contraction to explain the in vivo studies of bovine peptide-B in which the peptide behaves as a vasopressor substance producing vascular vasoconstriction. A possible mechanism for the participation of bovine peptide-B in the model for muscle contraction, based on the polarizing interaction of the highly charged density of negativity of the peptide with the actomyosin complex, is presented. Furthermore, bovine peptide-B is speculated as participating in vasoconstriction via attachment to some smooth muscle receptor.


Dimensional analysis of the left ventricle: effects of acute aortic regurgitation.

The effects of acute severe aortic regurgitation on the left ventricle were investigated in conscious, chronically instrumented dogs. Left ventricular dimensions and volumes were measured from biplane cineradiographs of beads positioned near the endocardium. Data were collected before and after the production of aortic regurgitation by a catheter technique. The aortic regurgitation resulted in increases in mean aortic pulse pressure from 44 to 73 mmHg (P smaller than 0.001), heart rate from 87 to 122 beats/min (P smaller than 0.02), and left ventricular end-diastolic pressure from 11 to 25 mmHg (P smaller than 0.05). Mean end-diastolic volume rose from 61 to 69 cc (P smaller than 0.001), while end-systolic volume remained unchanged at 37 cc. The end-diastolic dilatation following regurgitation was asymmetrical in that the increase in size was due principally to an increase in the septal-lateral axis. The acute volume load of aortic regurgitation was accomplished by an increase in end-diastolic volume, i.e., the Frank-Starling mechanism. The tachycardia probably reflects augmented cardiac sympathetic activity, but the constant end-systolic volume at a similar mean systolic pressure suggests that the net contractile state was unchanged.


Hypothalamic heating and cooling in monoamine-depleted rabbits.

The role of monoamines in the thermoregulatory responses induced by hypothalamic heating and cooling was investigated in conscious rabbits. Depletion of hypothalamic catecholamines by pretreatment with 6-hydroxydopamine (6-OHDA) greatly attenuated the rectal temperature and vasomotor responses to hypothalamic heating and cooling. Pretreatment with p-chlorophenylalanine (PCPA) depleted the animals of 5-hydroxytryptamine (5-HT). The results obtained are consistent with 5-HT having an inhibitory role in the rabbit's vasomotor response to heat stress and activating heat conservation during cold stress. Reserpinized rabbit were depleted of both norepinephrine (NE) and 5-HT and were found to be unresponsive to hypothalamic temperature changes. We conclude that the integrity of the monoaminergic system is viral for the correct functioning of the hypothalamus in maintaining a constant body temperature.


Guinea worm in southern Ghana: its epidemiology and impact on agricultural productivity.

In southern Ghana guinea worm disease was found to occur almost exclusively in villages dependent upon pond water during the dry season. The recent occurrence of guinea worm for the first time in many villages in the survey area suggests that the disease is spreading. The risk of increasing disease in the Accra plains is serious, because almost half of the 159 villages surveyed use pond water, and residents frequently travel to endemic areas. In this study adult male farmers were at greatest risk of becoming infected. The average work loss in untreated adults was more than 5 weeks. Because guinea worm disease is seasonal, conciding with peak agricultural activities, and few alternative labor sources are available for the incapacitated farmer, a marked reduction in agricultural output occurs. Additional research is needed to guide health education programs, to evaluate the effectiveness of chemical control of cyclops in ponds, and to develop low-cost improved rural water supplies.


Incipient atrophy of cornea. Experimental study.

The essence of the process in which the corneal tissue disappears in atrophy of the eye has not yet been studied. The authors of this investigation studied the changes in cornea as they developed after a gross perforating injury to the eye in rabbit. They found that the diameter of the cornea had grown smaller by 2 mm and its radius by 0.5 mm. The tensile strength of the cornea had decreased and hydration increased. Histologically there were but little changes and histochemically a decreased amount of acid mucopolysaccharides was ascertained in the corneal stroma. Incorporation of radioactive sulphate (Na2-35SO4) in vivo disclosed decreased biosynthesis of glycosaminoglycans. Clinically, even though the eye ball atrophied, complete atrophy with flattening and complete diminution, as in the human eye, never developed. The question arises whether the six-months period was sufficient for rabbit cornea to develop clinically similar atrophy as in human cornea or whether the clinical features are principally different in the two species. Foci of ossification, as found in atrophic eye balls, which are known to develop after a long time, would be evidence of the second alternative.


Corneal superficial unwettable defect. I. Clinical and histopathological study.

In 11 patients, the clinical, histological and histochemical picture of an unusual corneal affection termed "corneal superficial unwettable defect", is reported. It develops during the healing of various corneal lesions. Clinically, it is described as a defect of the epithelium with a denuded and changed Bowman's membrane, the surface of which repells tears and cannot be stained with fluorescein. In the histological picture, the changed Bowman's membrane has fibrillar or foamy appearence. It stains strongly with colloidal iron and alcian blue, but PAS and toluidine blue reactions are weak. Histochemical reactions for neutral fats are strongly positive, but those for phospholipids are negative. The results of extractions speak for the presence of some lipid component, which is neither neutral fat nor phospholipid. Hence it is supposed that there may be some waxes-ointment bases, reacting with the pathologically changed Bowman's membrane. The possible mechanisms injuring or influencing the Bowman's membrane are also discussed.


[Experience with using leukocytic interferon with methacil in influenza in children].

For prophylaxis influenza in the infection foci under stationary conditions 213 children were treated with leucocytic interferon with metacyl and 192 children were observed as the control group. The drug was administered intranasally in a dose of 0.25 ml once a day. The observations showed that among the children treated prophylactically with interferon and metacyl the number of influenza cases decreased 2.17 times a compared to the control group. In contact children who were taken ill with influenza the severety of the infection and the number of complications decreased. Under the stationary conditions in the children treated with interferon and metacyl the numbers of the wave-like infection and complications decreased 2.5 and 2.9 times respectively as compared to the children not treated with the drugs. Addition of metacyl to leucocytal interferon provided its administration once a day instead of 3-4 times a day.


Production of Sézary-like cells from normal human lymphocytes.

The present study was designed to determine if lymphocytes from healthy humans could be stimulated to assume the morphologic appearance of Sézary-like cells. Lymphocyte-rich populations of cells were incubated for 72 hours with pokeweed mitogen and phytohemagglutinin, both potent cellular mitogens. With light microscopy, differential cell counts performed on 0.5mu epoxy resin-embedded sections showed that 5% to 11% of the lymphocytes had cerebriform nuclei, and were designated as Sézary-like cells. The morphological results were confirmed by electron microscopy. Production of Sézary-like cells from stimulated, normal, human lymphocytes suggests that cells with cerebriform nuclei may represent reactive lymphocytes, and may explain their presence in benign inflammatory dermatoses.


Acne detergicans.

Popular brands of soaps and shampoos were assayed for comedogenicity in the rabbit ear. Only a small number produced hyperkeratosis when applied at a 25% concentration. Bacteriostatic substances, especially hexachlorophene, were mildly comedogenic. Conventional soaps include salts of fatty acids; the latter are known comedogens. The obsessive use of soaps by patients with acne vulgaris may aggravate the disease and result in its extension to unusual locations.


Nevoid follicular epidermolytic hyperkeratosis.

Comedo-like follicular papules of hair-follicles, confined to the left side of the upper part of the body, were found in a 22-year-old woman. Histological features were of epidermolytic hyperkeratosis with involvement of the follicular epithelium and the sebaceous duct with compact hyperkeratosis, but almost no interfollicular epidermal alterations were found. Because of the nevoid distribution, the prominent follicular plugging, and epidermolytic hyperkeratosis, we describe this condition as nevoid follicular epidermolytic hyperkeratosis.


Cutaneous hydroxystereoid dehydrogenases and rat-hair cycle.

The histochemical activity pattern of a number of hydroxysteroid dehydrogenases was studied during a provoked hair cycle in the rat skin. The intensity of reaction was high with dehydroepiandrosterone, estradiol, androsterone and pregnenolone, and none with 11-beta-hydroxyprogesterone, testosterone, dihydrotestosterone, 17-alpha-hydroxypregnenolone and 17-alpha-hydroxyprogesterone. Reactivity also varied with the hair cycle. The reactive sit-s were the acini of sebaceous glands and the hair sheaths.


Repeated topical applications of 1,2,4-trichlorobenzene. Effects on rabbit ears.

In a study to evaluate its acnegenic potential, increasing concentrations of 1,2,4-trichlorobenzene were applied topically to the ventral surface of the rabbit ear three times weekly for 13 weeks. Additional groups of rabbits received similar treatment with petroleum ether (solvent controls), received no treatment (negative controls), and received four once-weekly treatments with hexachlorodiphenyloxide, a known chloracnegenic agent (positive controls). Skin response to 1,2,4-trichlorobenzene was characterized grossly by dermal irritation directly related to the concentration of test material; there were the associated histologic changes of acanthosis and hyperkeratosis; there was no primary follicular involvement characteristic of acneform dermatitis. Dermal responses to hexachlorodiphenyloxide consisted of gross follicular enlargement, with waxy excretion on pressure, and severe scaling. The affected ear appeared thickened up to three times normal size and histologic sections showed primary follicular involvement characterized by marked thickening of the sheath and marked distention of the follicles with keratin, with resultant comedone formation, typical of chloracne.


Pulmonary microembolism associated with massive transfusion: I. Physiologic effects and comparison in vivo of standard and dacron wool (swank) blood transfusion filters in its prevention.

Pulmonary microembolism of microaggregates associated with massive blood transfusion may be a cause of post-traumatic pulmonary embolism. The purpose of this study was to investigate in the dog the influence on certain physiologic parameters of transfusion of blood containing platelet: white blood cell: fibrin (PWF) aggregates and to evaluate the effects of using blood transfusion filters of varying pore sizes during such transfusions. Exchange transfusions of approximately twice blood volume were performed in three groups of animals. Screen filtration pressure measurements verified the presence of large numbers of PWF aggregates in the transfusions. When no transfusion filters or standard commercially available blood transfusion filters of pore size 170 mu were used, experimental animals developed pulmonary hypertension, a decrease in total body 92 consumption, and metabolic acidosis. Interposition of Dacron wool (Swank) blood transfusion filters prevented these changes.


Conduit reconstruction of right ventricular outflow tract. Experience with 17 patients.

Evaluation was made of 17 patients who underwent conduit reconstruction of the right ventricular outflow tract (for anomalies other than truncus arteriosus) at the Texas Heart Institute between December, 1965, and June, 1974. Fifteen patients survived the operation and have shown substantial clinical improvement. Several different conduits were used, principally an allograft aorta with the aortic valve and mitral leaflet attached, a woven Dacron prosthesis containing a xenograft (porcine) valve, and a valveless Dacron tube graft. Allograft conduits are sometimes difficult to procure and keep and tend to calcify with passage of time. Woven Dacron prostheses are favored because they are readily available in a wide range of sizes. We believe it is not necessary for the conduit to contain a valve unless the patient has pulmonary hypertension, in which case we use a xenograft (porcine) valve because this valve does not require the long-term use of anticoagulants, a difficult regimen to manage in children. The largest possible prosthesis must be used; otherwise right ventricular hypertension will persist. Indications for conduit reconstruction include anomalous coronary arteries crossing the right ventricular outflow tract, discontinuity of the pulmonary arteries, and pulmonary atresia with a ventricular septal defect. Our current method of managing pulmonary atresia with ventricular septal defect (pseudotruncus arteriosus) includes palliative shunting to relieve hypoxemia during infancy and to permit full development of the pulmonary arteries for eventual total correction at a more optimal age when a larger conduit may be used.


Investigation of the nature of Australia antigen. I: The absence of biological activity of Australia antigen in human blood leukocyte culture.

Purified and concentrated preparations of Australia antigen had no stimulating effect on leukocytes of human subjects under study when tested either on DNA-polymerase activity, 3H-thymidine uptake or chromosomal alterations. Moreover, in patients with chronic hepatitis and cirrhosis of the liver no correlation between antigenemia and chromosome aberrations in blood leukocyte cultures could be detected. On the other hand, a serum obtained from a virus hepatitis patient with Australia antigen in the blood was found to stimulate leukocyte cultures from one patient with Down's syndrome and antigenemia, one mentally retarded patient and three normal donors. This stimulating agent is obviously not associated with Australia antigen.


Lipid disturbances associated with spiral muscular atrophy. Clinical, electromyographic, histochemical, and lipid studies.

Twelve patients with juvenile- and adult-onset spinal muscular atrophy have been studied. Eleven of the twelve patients had either type II, type IV, or borderline abnormal phenotypes, suggesting a possible relationship between serum lipid abnormalities and neuronal degeneration in the spinal muscular atrophies. Muscle enzyme histochemical studies provided valuable diagnostic information. Extensor toe signs and talipes cavus were common clinical observations.


Clindamycin. A Trojan horse?

Clindamycin is a widely used antibiotic with a spectrum that includes Gram-positive bacteria, with the exception of enterococci, and Gram-negative anaerobes. Toxicities include clindamycin colitis, a pseudomembranous colitis that can be fatal. The colitis usually is related to dosage and duration of therapy, but can occur after ingestion of only several capsules. Once diarrhea develops in a patient taking clindamycin, the drug should be stopped and diagnostic measures to define colitis initiated. If colitis is present, it should be managed aggressively, probably with corticosteroids and with intensive supportive measures.


Delay and block of cardiac impulse caused by enhanced phase-4 depolarization in the His-Purkinje system.

The underlying mechanism of bradycardia-dependent bundle-branch and paroxysmal atrioventricular block appears to be enhancement of phase-4 depolarization in a branch or in a natural or acquired monofascicular pathway. Clinical records of these forms of impaired conduction occurring in the bundle-branches, with either longer or shorter cardiac cycle lengths, are presented and analysed. These also include the combination of Mobitz typw I atrioventricular block with variable degrees of bundle-branch block, as a representative example of narrow ventricular escape beats firing in the zone where prominent diastolic depolarization is present.


Occurrence of coronary ostial stenosis in a necropsy series of myocardial infarction, sudden death, and violent death.

A postmortem coronary angiography technique employing aortic injection of contrast medium and double contrast visualization of the aortic bulb and large epicardial coronary trunks was applied to the study of coronary ostia in a series of 124 deaths from acute myocardial infarction and a series of 89 sudden deaths without recent infarction and 42 violent deaths. A stenosis of 50 per cent or more of the lumen was found in the right ostium in 45 per cent and in the left ostium in 8 per cent of infarct cases. The corresponding figures in sudden deaths were 37 per cent on the right and 4.5 per cent on the left side, and in violent deaths 7 per cent in the right ostium and none in the left. Most ostial stenoses were caused by coronary atherosclerosis. In 9 patients, two with a recent infarct and 7 sudden deaths, an ostial stenosis was the only stenosed site in the coronary arterial tree. Of theses 9 patients, 7 were known to have suffered from symptomatic heart disease during life, chest pain on effort and arrhythmias being the most common complaint.


Electrocardiogram of secumdum type atrial septal defect simulating endocardial cushion defect.

Three groups of patients are identified in the series of secumdum atrial septal defect with left axis deviation in an analysis of 910 patients of secundum atrial septal defect with or without other accompanying anomalies: 1) 12 patients with isolated ostium secumdum defect; 2) 5 patients with associated prolapse of posterior leaflet of the mitral valve; 3) 4 patients with associated hypertrophic cardiomyopathy and/or single (left) coronary artery. The findings of these 21 patients with proved secumdum atrial septal defect are analysed. The combination of such anomalies should be considered in the differential diagnosis of primum endocardial cushion defect from the electrocardiographic viewpoints.


Effect of intermittent left anterior hemiblock on left ventricular function.

Studies of left ventricular function were performed during intermittent left anterior hemiblock, Max dP/dt and Vmax fell during left anterior hemiblock, with a rise in left ventricular end-diastolic pressure compared with a normally conducted beat.


An equilibrium binding study of the interaction of fructose 6-phosphate and fructose 1,6-bisphosphate with rabbit muscle phosphofructokinase.

Equilibrium binding studies of the interaction of rabbit muscle phosphofructokinase with fructose 6-phosphate and fructose 1,6-bisphosphate have been carried out at 5 degrees in the presence of 1-10 mM potassium phosphate (pH 7.0 and 8.0), 5 mM citrate (pH 7.0), or 0.22 mm adenylyl imidodiphosphate (pH 7.0 and 8.0). The binding isotherms for both fructose 6-phosphate and fructose 1,6-bisphosphate exhibit negative cooperativity at pH 7.0 and 8.0 in the presence of 1-10 mM potassium phosphate at protein concentrations where the enzyme exists as a mixture of dimers and tetramers (pH 7.0) or as tetramers (pH 8.0) and at pH 7.0 in the presence of 5 mM citrate where the enzyme exists primarily as dimers. The enzyme binds 1 mol of either fructose phosphate/mol of enzyme monomer (molecular weight 80,000). When enzyme aggregation states smaller than the tetramer are present, the saturation of the enzyme with either ligand is paralleled by polymerization of the enzyme to tetramer, by an increase in enzymatic activity and by a quenching of the protein fluorescence. At protein concentrations where aggregates higher than the tetramer predominate, the fructose 1,6-bisphosphate binding isotherms are hyperbolic. These results can be quantitatively analyzed in terms of a model in which the dimer is associated with extreme negative cooperativity in binding the ligands, the tetramer is associated with less negative cooperativity, and aggregates larger than the tetramer are associated with little or no cooperativity in the binding process. Phosphate is a competitive inhibitor of the fructose phosphate sites at both pH 7.0 and 8.0, while citrate inhibits binding in a complex, noncompetitive manner. In the presence of the ATP analog adenylyl imidodiphosphate, the enzyme-fructose 6-phosphate binding isotherm is sigmoidal at pH 7.0, but hyperbolic at pH 8.0. The characteristic sigmoidal initial velocity-fructose 6-phosphate isotherms for phosphofructokinase at pH 7.0, therefore, are due to an heterotropic interaction between ATP and fructose 6-phosphate binding sites which alters the homotropic interactions between fructose 6-phosphate binding sites. Thus the homotropic interactions between fructose 6-phosphate binding sites can give rise to positive, negative, or no cooperativity depending upon the pH, the aggregation state of the protein, and the metabolic effectors present. The available data suggest the regulation of phosphofructokinase involves a complex interplay between protein polymerization and homotropic and heterotropic interactions between ligand binding sites.


Rat testis mitochondrial adenylate cyclase. Partial purification and characterization.

1. Adenylate cyclase (EC 4.6.1.1) from rat testis mitochondria has been solubilized by treatment with the non-ionic detergent Lubrol PX. The soluble enzyme was further purified by DEAE-cellulose chromatography. 2. The specific activity of the adenylate cyclase eluted from the DEAE-cellulose column was found to be four times higher than that of an intact mitochondrial preparation. At this step the enzyme shows a sedimentation coefficient of 4.2 S and a diffusion coefficient (D) of 3.12 - 10- minus 7 cm-2/sec. 3. Solubilization of the adenylate cyclase resulted in loss of responsiveness to gonadotrophic hormones. Addition of phosphatidylserine to the soluble preparation partially restored the activation of adenylate cyclase by human chorionic gonadotrophin. 4. The results of this study suggest that the activity of the adenylate cyclase may be dependent on the membrane-bound phospholipids and that the enzyme attached to the mitochondrial membranes has some properties which are similar to the adenylate cyclase found to be associated with other membrane systems of the cell.?


Effects of N-acetylimidazole on human erythrocyte ATPase activity. Evidence for a tyrosyl residue at the ATP binding site of the (Na+ plus K+)-dependent ATPase.

1. Acetylation of human erythrocytes by N-acetylimidazole alters the structure of stroma prepared from these cells and the degree of alteration appears to be dependent upon the level of the initial treatment. These changes do not occur when stroma are acetylated. 2. Deacetylation by hydroxylamine or mild alkaline treatment causes a complete recovery of the (Na+ plus K+)-dependent and the Ca2+ -stimulated ATPase activities and indicates that the inhibition is due to the acetylation of a tyrosyl residue. There is only partial recovery of the Mg2+ -dependent ATPase after deacetylation. 3. ATP or Mg-ATP completely protect the (Na+ plus K+)-dependent ATPase, but not the Ca2+ -stimulated system. 4. The results indicate that the (Na+ plus K+)-dependent and the Ca2+ -stimulated ATPase activities have separate substrate binding sites and most likely are separate enzyme systems. 5. Acetylation of human erythrocytes has no effect on D-glucose transport.


Energy-dependent endocytosis in erythrocyte ghosts. IV. Effects of Ca2+, Na+ +K+, and 5'-adenylylimidodiphosphate.

The requirement of actual splitting of ATP for endocytosis in erythrocyte ghosts has been confirmed by use of the ATP analog, 5'-adenylylimidodiphosphate, (AMP-P(NH)P). This compound, in which the oxygen connecting the beta and gamma phosphorus atoms was replaced by an NH group, did not cause endocytosis nor was it a substrate for ATPase activity. AMP-P(NH)P was a competitive inhibitor both for the endocytosis and the Mg2+-ATPase activities. The K1 of AMP-P(NH)P for Mg2+ ATPase activity was 2.0 - 10-4 M and, while the Km of ATP for this activity was also 2.0 - 10-4 M indicating nearly identical affinities of ATP and AMP-P(NH)P for the active site. ADP, or ADP plus orthophosphate, did not cause endocytosis, showing that endocytosis was not due to binding of the products of ATP hydrolysis. Sodium or potassium ion or ouabain had no effect on endocytosis, which eliminated the possibility of involvement of the Na+, K+ ATPase in the endocytosis process. Calcium could not be substituted for magnesium; rather it inhibited endocytosis at the concentration of 1 - 10-3 M. EGTA relieved the inhibitory effect of Ca, which indicated that the binding of calcium to the membrane was reversible. These experimental results reaffirm the conclusion that ATP must be split to engender endocytosis under these conditions. Some characteristic parameters of the hemoglobin-free porcine erythrocyte ghosts were studied in order to characterize the system more adequately.


Surgical management and problems of heart pacing.

Experience with 40 patients and 150 pacemakersss is reviewed. In 13 patients the initial pacing was by epicardial electrodes and in 27 by transvenous catheter electrodes. When epicardial systems were used, two electrodes were always fixed to the heart, the additional electrode being provided for temporary pacing and as a spare in case of future need. The battery unit was usually buried behind the rectus abdominis muscle. Vacuum drainage of the pacemaker pocket was used and was considered helpful in avoiding some complications. The right cephalic vein was preferred for transvenous pacing but existing venous abnormalities sometimes required use of the external jugular vein. Five patients had epicardial systems changed to transvenous ones, and in 5 the reverse procedure was needed. Pacemaker battery problems included inadequate soft tissue protection for the unit, epoxy resion fracture, titanium case leak and problems relating to design change and the special needs for overseas travel. While the danger from diathermy and radiofrequency is now recognized, similar dangers exist for those paced by 'demand' units in large electromagnetic fields.


An abnormality in sebaceous function in phenylketonuria.

The sebum excretion rate has been found to be increased in female patients with phenylketonuria. This may be related to a depletion of midbrain dopamine and release of the sebotrophic hormone.


The density of synapses and neurons in normal, mentally defective ageing human brains.

The densities of synapses and neurons were measured in 7 neurosurgical specimens of cerebral cortex, regarded as normal controls. The average result was 6 times 10-11 synapses/cm3, 15-6 times 10-6 neurons/cm3, and 38,000 synapses per neuron. Specimens of frontal and temporal cortex from 7 ageing brains (68 to 89 years) gave closely similar counts of neurons and synapses. Seven specimens from 3 severely retarded brains gave similar counts of neurons and slightly higher counts of synapses. A deficit of synaptic development is thus not a necessarre discussed.


Molecular and biological studies on cardiac muscle calcium-binding protein (TN-C).

TN-C was purified from bovine cardiac muscle. In the absence of Ca-2+, cardiac TN-C has an intrinsic sedimentation coefficient of 1.93 S and a molecular weight of 18 000 daltons. Cardiac TN-C reverses the inhibitory effect of skeletal TN-I on the Mg-2+-activated ATPase of a skeletal synthetic actomyosin preparation in the presence of skeletal tropomyoson. Circular dichroism (CD) studies indicate that cardiac TN-C undergoes a major conformational change upon binding Ca-2+. A similar response is elicited by Sr-2+, whereas Mg-2+ has a much less pronounced effect. The presence of Mg-2+ does not alter the net effects of either Ca-2+ or Sr-2+. Cardiac TN-C is rich in acidic amino acid residues. UV absorption, near UV CD, and fluorimetric studies show that the protein lacks tryptophan and has a relatively high phenylalanine to tyrosine ratio. The results of this study invite direct comparisons with results reported for the skeletal muscle analogue of cardiac TN-C.


Participation of exogenous thymine and thymidine in deoxyribonucleic acid synthesis in Lactobacillus acidophilus.

The degree of participation (DP) of exogenous thymidine and thymine in overall DNA synthesis was studied in Lactobacillus acidophilus R-26. The DP of thymidine remains constant under a variety of conditions (except at low thymidine concentrations, when the DP is influenced by deoxyribonucleosides and folic acid). A 5-bromodeoxyuridine-resistant mutant was isolated, which displayed cross-resistance to 5-bromouracil and a significantly lower DP of thymidine than the parental strain. Thymine was poorly incorporated in the parental strain even in the presence of deoxyribosides. The results of this investigation would be compatible with the possibility of an alternative pathway for thymidylate synthesis other than the known thymidylate synthase pathway.


Thymic origin of abnormal lymphoid cells in Sézary syndrome.

A patient with a 5-year history of pruritus and progressive and generalized erythroderma was found to have abnormal lymphocytes in the peripheral blood and mononuclear dermal infiltrate, all of which are features consistent with those described in Sézary syndrome. Systemic chemotherapy produced an almost complete resolution of skin lesions and pruritus. Serial studies on the abnormal cells included responses to phytohemagglutinin, cytogenetics, and immunofluorescence tests for identification of B or T lymphocytes. The abnormal cells demonstrated hyperdiploidy; the ratio of B to T cells fell whenever abnormal lymphocytes appeared in the peripheral blood, and returned to normal when abnormal cells disappeared from circulation. Lymphocytes separated from a skin nodule labeled as T cells. We conclude that the abnormal lymphocytes in this patient are of thymic origin.


Membrane glycoprotein differences between normal lactating mammary tissue and the R3230 AC mammary tumor.

Membrane glycoproteins have been studied in the normal lactating mammary gland and R3230 AC mammary tumor of the rat. Plasma membrane-enriched fractions were obtained from these tissues by discontinuous sucrose gradient centrifugation of a microsomal preparation from the tissue homogenates. The lightest membrane fractions (F-1 and F-2) have the greatest enrichment of plasma membrane markers, with a 14- to 20-fold purification of 5'-nucleotidase and Na+-K+ -adenosine triphosphatase over the homogenate values in both tumor and normal tissues for F-1. Electron microscopy shows smooth membrane vesicles for these fractions. Polypeptide analysis by acrylamide gel electrophoresis shows essentially the same patterns for F-1 and F-2 and only relatively minor differences between membrane components of tumor and normal tissues. Glycoprotein analysis of the polyacrylamide gels by periodate-Schiff staining indicates more dramatic differences. Membrane Fraction F-1 from normal tissue contains two major glycoproteins, GP-II and GP-III, while Fractions F-2 and F-3 contain an additional glycoprotein, GP-I, with a higher apparent molecular weight. In the tumor, the component corresponding to GP-III is decreased or absent and a new component GP-IV is seen at a lower apparent molecular weight.


Estriol prevention of mammary carcinoma induced by 7,12-dimethylbenzanthracene and procarbazine.

The concentration of estrogenic, androgenic, progestational, and adrenocortical steroid hormones in body fluids of mature intact Sprague-Dawley female rats was increased by s.c. implantation of 5 to 7 mg NaCl pellets containing 1 to 20% steroid 48 hr before administration p.o. of either 7,12-dimethylbenz(a) anthracene or procarbazine. The incidence of rats developing one or more mammary carcinomas in each treated group was compared to that ovserved in simultaneously treated groups receiving only the carcinogen, steroid, or no treatment whatsoever, with weekly observation of all rats until palpably growing tumors were biopsied and proven carcinomatous or until death occurred from other causes determined by autopsy. A total of 105 untreated or steroid-implanted rats followed to death (234 to 256 days median observation) developed no breast carcinomas. Rats fed either of the carcinogens developed initial evidence of breast carcinoma, after 136 to 156 days median observation, in 51 to 57% of 318 total treated rats. Nonbreast carcinomas and sarcomas developed in 5 to 10% of the carcinogen-treated rats.


Variable effects of heart rate on phasic and regional left ventricular muscle blood flow in anaesthetized dogs.

Tachycardia to 240 beats/min in anaesthetized dogs increases coronary blood flow while flow in the left ventricular wall remains homogeneous. An arteriovenous fistula causes subendocardial ischaemia at 120 beats/min but pacing to 180 to 210 beats/min increases subendocardial blood flow by increasing diastolic coronary driving pressure per minute.


Haemodynamic changes in adrenaline-induced acute massive lung oedema.

During the production of adrenaline-induced acute massive lung oedema in the dog, plumonary arterial, pulmonary venous, systemic arterial, and bronchial arterial blood pressures all increase markedly. Pulmonary arterial and venous blood flows fall steeply after initial transient rises. Systemic arterial blood flow also declines, with or without an initial transient increase. The bronchial arterial blood flow shows an initial fall followed by a rise of late onset. The main determinant for the pathogenesis of adrenaline-induced lung oedema is apparently the enormously increased hydrostatic pressure in the pulmonary vascular bed.


The effect of cardiac tamponade on coronary haemodynamics in the awake dog.

Phasic left circumflex coronary artery and aortic blood flow were monitored in six awake dogs during a control period and at several degrees of cardiac tamponade. A mean pericardial pressure of 24 plus or minus 3 mm Hg (mean plus or minus SEM) was attained at the maximum tamponade level. Total left circumflex coronary blood flow decreased 51% while the systolic portion of this flow became negative or retrograde. Following acute relief of the tamponade, a coronary hyperaemic response was noted. It it suggested that myocardial ischaemia may be partially responsible for the depressed cardiac function seen in this condition and that extravascular compression of the epicardial vessels may limit the coronary blood flow during systole.


Structure and innervation of the pineal gland of the rabbit, Oryctolagus cuniculus (L.). III. An electron microscopic investigation of the innervation.

In the rabbit pineal gland two types of postganglionic nerve endings were found which are characterized by the presence of small dense-core vesicles or small clear vesicles. Pharmacological and cytochemical experiments showed then to be noradrenergic and cholinergic, respectively. Both types were often present in the same nerve bundle, occasionally in close opposition. Intrapineal neurons were only rarely observed. They showed cholinergic synapses on their perikaryon and dendrites as well as noradrenergic axo-dendritic close contacts. Bilateral extirpation of the superior cervical ganglia revealed the postganglionic sympathetic origin of the pineal noradrenergic nerve fibres. Moreover, it appeared that these ganglia are hardly, if at all, involved in the pathway of pineal cholinergic innervation. The results obtained from lesions of both facial nerves, taken together with the results reported in the literature, led to the conclusion that the postganglionic cholinergic nerve fibers in the pineal are of parasympathetic origin. A model for the sympathetic and parasympathetic pineal innervation is proposed.


The location of proteins labeled by the 125I-lactoperoxidase system in the NIL 8 hamster fibroblast.

NIL 8 hamster fibroblast cells were labeled by lactoperoxidase-catalyzed iodination. Their membranes were fractionated by sedimentation-rate and isopycnic zonal centrifugation. All the iodinated proteins except the very prominently labeled high molecular weight protein (greater than 200,000 daltons) were located in a fraction identified enzymically and compositionally as plasma membrane. The high molecular weight protein that was previously shown to be sensitive to virus transformation (Hynes, 1973) is concentrated in a very high density particle (rho equals 1.253-1.259) which contains mainly carbohydrate and protein and very low levels of lipid. 5'-nucleotidase was the only enzyme reproducibly demonstrated in this fraction, and electron micrographs revealed a predominantly amorphous morphology together with a few membraneous structures. The iodine label in this fraction was very sensitive to trypsinization prior to homogenization. All the available evidence indicates that this fraction is derived from the surface coat. Mitochondria, nuclei, and soluble protein were labeled to an insignificant extent. The presence of the iodinated surface proteins associated with the endoplasmic reticulum fraction is discussed in the light of these results.


Effects of 5,5'-diphenyl-2-thiohydantoin on respiration and oxidative phosphorylation of rat liver mitochondria.

5,5'-Diphenyl-2-thiohydantoin (DPTH) administered in vitro, inhibited state 3 oxidation, stimulated state 4 oxidation and decreased ADP:O ratio when 3-hydroxybutyrate and succinate were used as substrates. Considerably lower DPTH concentrations were required for the inhibition of 3-hydroxybutyrate oxidation (50% inhibition occurred at approximately 0.17 mumoles DPTH/mg protein) than were needed for inhibition of succinate oxidation (50% inhibition occurred at about 0.62 mumoles DPTH/mg protein). DPTH showed no inhibitory effects when ascorbate plus tetramethylphenylenediamine (TMPD) served as the substrate. The inhibition of state 3 respiration was not reversed by 2,4-dinitrophenol (DNP), although there was a slight increase in the DNP rate:state 3 rate suggesting the presence of a weak DPTH inhibotory site located within the Site I energy transport chain. Uncoupling, in the presence of DPTH, was observed with all substrates. In experiments utilizing sonicated mitochondria, DPTH inhibited NADH-linked oxidation, but did not inhibit succinate or ascorbate plus TMPD oxidation. The effects of DPTH were reversed by dilution and by addition of albumin. DPTH concentrations which produced inhibition of state 3 respiration in vitro were reached, in vivo, in the livers of rats receiving a single oral dose of 40 mg/kg of DPTH.


Seven-year follow-up of cardiovascular study and maximal exercise of Chinese men.

A seven-year follow-up in 1973 of a prospective cardiovascular study of 1820 initially, healthy, middle-aged Chinese men of 40-59 years of age identified 1745 (95.9%) known survivors, 49 (2.7%) interim deaths, and 26 (1.4) who could not be traced. Of the survivors, 1462 (83.8%) were re-examined, 292 (16.7%) had another treadmill test of maximal exercise, and 283 (16.2%) failed to return for re-examination. On the basis of interim surveillance of hospital admissions, questionnaires and re-examination, a greater incidence of noncardiovascular events (338 or 18.6%) than evidence of cardiovascular disease (220 or 12.1%) was found while the majority (1021 or 56.1%) remained healthy. Total mortality was 0.29 for men under 50 and 0.76 per 100 person-years for men of 50 or more years of age. Only nine, or 18.4% of the deaths were due to cardiovascular causes, and unexpectedly for this population sample, only three were attributed to stroke. When cardiovascular morbidity was related to presence of ST depression after maximal exercise, to hypertension at rest by WHO criteria, to both findings, or to absence of either on initial intake examination, incidence increased from 2.3% for NEITHER group, to 5.7% for ST group, to 11.9% for HT group, and to 25.0% for BOTH groups. Re-examination revealed more evidence of cardiovascular disease than did surveillance of hospital admissions. Additional to effects of aging and mild adiposity, longitudinal changes in blood pressure and ST depression, increasing in the NEITHER group, but less frequent in the other groups, showed some evidence of regression toward the mean, as well as emerging disease and the confounding effects of uncontrolled treatment of hypertension in many. The potential for prediction of subsequent cardiovascular morbidity or mortality appeared stronger for hypertension than for postexertional ST depression, although the two were additive in this population, which is more prone to hypertension and stroke but now is developing clinical manifestations of coronary heart disease more frequently.


The spectrum of urinary basement membrane antigen excretion in normal and pathological rabbit urine.

The work presented here concerns the excretion of basement membrane-like fragments into the urine of rabbits. The spectrum of urinary antigen excretion in normal rabbits as well as those with Masugi nephritis and acute serum sickness was investigated. It was found that the urinary antigen content of rabbits with Masugi nephritis was markedly different to that of normal urine whereas urine from rabbits with acute serum sickness was similar. This difference between the two groups was presumably due to the more extensive renal lesions seen in Masugi nephritis compared to the mainly glomerular lesions of acute serum sickness. It was concluded that studies on urinary basement membrane antigens can provide information on the severity and extent of renal lesions.


1.3 Bis-(2 chloroethyl)-1-nitrosourea and streptozotocin chemotherapy.

Streptozotocin (STZ) was combined with 1,3 bis-(2 chloroethyl)-1-nitrosourea (BCNU) and with BCNU and 5-fluorouracil (FU) in a 2- and 3-drug clinical chemotherapeutic trail. The premise that STZ and BCNU are qualitatively different with regard to marrow suppression was the primary rationale of the study. The 2- and 3-drug regimes were associated with a higher incidence of severe leukopenia and thrombopenia (47% and 100%, respectively) and a lower mean nadir for each (1,700/mm3 and 15,000/mm3) than the reported experience with single drug BCNU therapy. However, this synergism did not apply to therapeutic effects. The reasons for potentiation of marrow toxicity may be related to specific aspects of direct drug interaction as well as alterations in pharmacologic reactions.


[Prognosis of cirrhosis of the liver (15-year follow-up from time of first manifestation (author's transl)].

114 patients with cryptogenic or "hepatitic" cirrhosis of the liver (toxic or biliary causes excluded) were followed for a period of 15 years. The time of first manifestation of the disease was known because all patients had been in hospital or under serial outpatient observation, with clinical and biopsy studies every six months for chronic hepatitis. In addition, diagnosis of the liver cirrhosis was confirmed by laparoscopy and histomorphology, which further confirmed the exact timing of first manifestation. In 37 patients (32.5%) the disease healed out completely by way of reactionless fibrosis after, in some of them, a long period of observation, while in 47 (41.2%) the active disease continued, and 30 (26.3%) died in liver coma or bleeding from varices or surgical shunt procedures or from carcinoma of the liver. Prognosis of cirrhosis of the liver is thus much better than previously thought.


Effects of somatostatin and hypothalamic ventromedial lesions on GH release induced by morphine.

Morphine in intravenous doses ranging from 10 mug/kg to 8 mg/kg was shown to be effective in stimulating GH release in the unanesthetized rat. The response to the log of the dose was linear over a range of 10 to 1000 mug/kg. Somatostatin (GH-release inhibiting factor) administered SC in a dose of 200 mug/kg 5 min before morphine prevented the GH rise. Neither inhibitors of catecholamine or serotonin synthesis nor blockage of alpha and beta-adrenergic receptors had any effect on the response. The response was partially blocked in animals with large hypothalamic ventromedial (VMN) lesions. Such lesions completely abolished the GH response to pentobarbital. These results indicate that morphine is a remarkably potent agent for stimulation of GH release but the precise mechanism and site of action of the drug remain to be determined.


The hypothalamo-neurohypophysial complex in organ culture: morphologic and biochemical characteristics.

Organ cultures of the guinea pig hypothalamo-neurohypophysial complex could be maintained for periods as long as 3 weeks. Morphological studies using conventional light microscopy as well as electron microscopy indicate that neurosecretory cells remain viable during this time. Biochemical studies show that the capability of the cultures to synthesize a normal spectrum of cellular RNA species is impaired during the initial day in culture, but that this capability is restored after 4-5 days. Similarly, protein synthesis proceeds at low levels during initial days of culture, but increases after 5 days. These phenomena appear to be independent of changes in the specific radioactivity of precursor pools and were also observed when outgrowth of non-neuronal cells is inhibited with fluorodeoxyuridine. The content of vasopressin, a product of a specific class of neurosecretory neurons, was found to decrease in the posterior pituitary during 7 - 10 days in culture by 50-70 percent; the levels then plateaued and were maintained for up to 3 weeks. The hypothalamic content of the hormone was relatively constant throughout the culture period. Of most importance was the demonstration that the organ cultures were capable of vasopressin biosynthesis. This capability paralleled the biosynthetic activity of RNA and gross protein metabolism in that there was an initial refractory period of several days duration.


Effects of aminergic drugs and glutamic acid on audiogenic seizures induced by early exposure to ethanol.

Adult C57BL/10Bg mice, normally resistant to audiogenic seizures, became susceptible when the mothers drank 10 per cent ethanol in water during pregnancy and for 14 days postpartem. Reserpine enhanced the incidence of seizures, and the effect was reversed by 5-hydroxytryptophan but not by dihydroxyphenylalanine. p-Chlorophenylalanine also enhanced the incidence of seizures, whereas alpha-methyl equals p equals tyrosine did not effect. Monsodium glutamate almost completely prevented seizures. These results are consistent with the interpretation that the serotonergic systems may be among those involved in the seizure mechanism induced by fetal and early exposure to ethanol.


Influence of non-H-2 genotype on the response to H-2-linked mixed lymphocyte reaction stimulating (MLR-S) genes.

The intensity of the mixed lymphocyte reaction to H-2-linked mixed lymphocyte reaction stimulating genes is influenced by non-H-2 genotype. The same H-2 incompatibility results in a stronger mixed lymphocyte reaction when both partners have the C57BL/10ScSnPh genetic background than when they have the genetic background of the A/Ph strain. It is suggested that, as in man, the intensity of the mixed lymphocyte reaction to incompatibility at the MLR-S genes linked to the major histocompatibility complex is controlled by MLR-response genes not linked to MHS.


Immunoperoxidase demonstration of human serum globulin binding to islet tissue.

Sera from juvenile diabetic and nondiabetic controls were tested, using the immunoperoxidase method, for the presence of islet-binding immunoglobulins. All diabetic sera tested contained an islet-binding protein and on the average the sera were more positive than age-matched controls. Normal adult sera are undifferentiated from juvenile diabetic sera. Most sera from children less than two years of age did not bind to islet tissue and sera from cystic fibrosis patients had a markedly diminished ability to bind to islet tissue. The binding protein appears to be an immunoglobulin which selectively reacts with elements of the islet beta cell.


The effect of dextran infusions on glycosaminoglycan excretion in the Sanfilippo syndrome.

Infusion of a dextran solution of equivalent osmolality to plasma into a patient with the Sanfilippo syndrome caused an increased excretion of uronic acid. This increase was less than that obtained previously, when an equal amount of plasma was infused into the same patient. Moreover, there were no significant changes in the ratio of larger to smaller glycosaminoglycan fragments or in the sulphate/uronic acid ratios, in contrast to the changes noted subsequent to plasma infusion. It is concluded, therefore, that in this case infusion of dextran was not as effective as plasma in inducing a temporary increased mobilisation of pathological glycosaminoglycans.


The significance of motor handicap in the prognosis of childhood epilepsy.

A total of 244 epileptic children were collected in an epidemiological and prognostic investigation on epilepsy. The children were divided into four groups according to their motor performance. There were 150 children with no motor handicap, 32 with clumsiness, 51 with cerebral palsy and 11 with severe muscular hypotonia associated with grave mental handicap. A study was made of age at onset of epilepsy, intelligence level, maximum frequency of seizures, grand mal status, results of medical treatment, and the time elapsed since the last seizure. A significant correlation was found between severity of motor handicap and poor prognosis of epilepsy.


Immunogenicity of an aerogenic BCG vaccine in T-cell-depleted and normal mice.

Aerogenic infection of adult thymectomized, lethally irradiated, bone marrow-reconstituted (THXB) C57B1 times C3H F1 hybrid mice with 1 to 3,000 viable BCG Montreal was followed by an extended period of logarithmic growth to a maximum population of 5 times 10-6 bacilli by day 35. The infection spread to the liver, spleen, and bone marrow with extensive multiplication in all test organs before the growth curves abruptly entered a stationary phase. Up to 30% of the THXB mice eventually died as a result of the ongoing BCG infection. There was no sign of an antimicrobial immune response in the THXB mice analogous to that seen in the control animals beginning about day 30. The THXB mice developed considerable immediate but no delayed hypersensitivity to PPD. Intravenous challenge of the BCG-vaccinated THXB mice with 105 virulent Mycobacterium tuberculosis Erdman indicated that they were as susceptible to the tuberculous challenge as a group of unvaccinated controls. Visible surface lesions developed on the lung 90 days postinfection in the T-cell-depleted host with a sharp rise in counts to 175 per lobe on day 120 followed by a plateau for the remainder of the study. Control mice developed visible lesions about  day 50, with 225 lesions per lobe by day 70 and a sharp decline to undetectable levels by day 90. The histopathology of these changes was examined carefully, together with the rate of cellular proliferation (tritiated thymidine uptake) by lung and spleen cells as the BCG infection progressed in the THXB mice. Peak uptake by both organs was depressed during the early stages of the BCG infection in the T-cell-depleted mice, but later the incorporation rates were significantly elevated above control values as the infection progressed.


Helminth parasitisms among intermingling insular populations of white-tailed deer, feral cattle, and feral swine.

Helminth infections among free-ranging, intermingling populations of white-tailed deer (Odocoileus virginianus), cattle (Bos taurus), and swine (Sus scrofa) on an island off the Georgia coast were studied. Of 39 species of helminths collected, 19 were found in deer, 17 in cattle, and 13 in swine. Of 28 species of helminths recovered from ruminants, 8, viz, Capillaria bovis, Cooperia punctata, Dictyocaulus viviparus, Gongylonema pulchrum, G verrucosum, Haemonchus contortus, Moniezia benedeni, and Trichostrongylus axei, occurred in both deer and cattle. Common liver flukes (Fasciola hepatica) infected cattle and swine but not deer. Only 1 helminth, G pulchrum, infected deer, cattle, and swine. The findings suggested that helminths harbored by the host species are distinct, with little exchange occurring.


Effects of ribonuclease A on amino acid transport in Neurospora crassa.

Incubation of Neurospora crassa conidia with ribonuclease (RNase) A reduces transport of L-phenylalanine by those cells. Under similar conditions, oxidized RNase A, RNase T1, and RNase T2 do not have this effect. Incubation of conidia with active RNase covalently attached to polyacrylamide beads reduces L-phenylalanine transport. This indicates that the site of enzymatic action is at the cell surface. At the lower concentration of enzyme used in this study, incubation with RNase A reduces transport of L-phenylalanine by the general (G) amino acid permease. Increasing the enzyme concentration results in reduction of transport by the neutral aromatic (N)-specific permease. The increased transport activity that accompanies onset of conidial germination is also sensitive to incubation with RNase A. Application of the enzyme to actively transporting cells does not release amino acid transported prior to enzyme addition. Cells cultured on media supplemented with [2-14C] uridine release isotopic activity after RNase A incubation. Analogous treatments with Pronase, RNase T1, RNase T2, or deoxyribonuclease I do not release isotope activity. Pronase treatment does reduce L-phenylalanine transport. Incubation of conidia with RNase A also inhibits germination of those conidia.


Characterization of Neurospora crassa mutants deficient in glucosephosphate isomerase.

Two independent mutants of Neurospora crassa lacking glucosphosphate isomerase activity (gpi) were isolated. These mutants were obtained as double mutants containing the pp or T9 mutation in addition to the gpi mutation located on linkage group IV; the pp mutation caused the inability to form protoperithecium and the loss of ascospore germination, and the T9 mutation caused the alteration in glucoamylase and several growth characteristics. The gpi mutants did not grow on fructose but grew on glucose or sucrose. Growth of these mutants on glucose was stimulated by addition of fructose. The gpi mutants showed restricted colonial growth on agar media containing glucose in contrast to the normal filamentous growth of the wild-type stain.


Isolation of extrachromosomal deoxyribonucleic acid for exfoliative toxin production from phage group II Staphylococcus aureus.

The ability of phage group II staphylococcal strain UT 0101 to produce exfoliative toxin and bacteriocin could be eliminated at a high frequency after growth at high temperatures or in the presence of ethidium bromide or sodium dodecyl sulfate. Extrachromosomal deoxyribonucleic acid, associated with the genes for exfoliative toxin and bacteriocin production, was isolated from strain UT 0101 but was absent from an ethidium bromide-cured substrain. The molecular weight of the exfoliative toxin plasmid, determined by co-sedimentation with the penicillinase plasmid, PI258, was 3.3 times 10-7. The 56S covalently closed circular form of the exfoliative toxin plasmid converted to a 38S open circular form after storage or exposure to sodium dodecyl sulfate. Plasmid deoxyribonucleic acid associated with penicillin resistance could not be identified in the penicillin-resistance Tox+ strains, UT 0007 and UT 0001.


The subunit structure of tryptophan synthase from Neurospora crassa.

Tryptophan synthase of Neurospora crassa was purified to electrophoretic homogeneity from the wild type strain 74A which had been derepressed by the presence of 0.5 mM indoleacrylic acid in the growth medium. The isolated material migrated as a single symmetrical peak in the ultracentrifuge with a sedimentation constant of 6.0 S. Gel filtration on Sephadex G-200 AND CONVENTIONAL SEDIMENTATION EQUILIBIRIUM YIELDED MOLECULAR WEIGHT ESTIMATES OF 151,000 PLUS AND MINUS 10,000 AND 149,000 PLUS AND MINUS 10,000, RESPECTIVELY. Treatment of the enzyme with sodium dodecyl sulfate followed by polyacrylamide gel electrophoresis gave a single band with a relative mobility suggesting a molecular weight of 76,000 plus and minus 2000. Aspartic acid was the only detectable NH2-terminal amino acid and experiments with carboxypeptides A and B revealed that the three amino acids, isoleucine, leucine, and phenylalanine, were released rapidly and in the order mentioned. These results are interpreted as indicating that the Neurospora enzyme is a homodimer.


Synthesis of adenosine triphosphate and exchange between inorganic phosphate and adenosine triphosphate in sodium and potassium ion transport adenosine triphosphatase.

Radioactive adenosine triphosphate was synthesized transiently from adenosine diphosphate and radioactive inorganic phosphate by sodium and potassium adenosine triphosphatase from guinea pig kidney. In a first step, K+-sensitive phosphoenzyme was formed from radioactive inorganic phosphate in the presence of magnesium ion and 16 mM sodium ion. In a second step the addition to the phosphoenzyme of adenosine diphosphate with a higher concentration of sodium ion produced adenosine triphosphate. Recovery of adenosine triphosphate from the phosphoenzyme was 10 to 100% in the presence of 96 to 1200 mM sodium ion, respectively. Potassium ion (16mM) inhibited synthesis if added before or simultaneously with the high concentration of sodium ion but had no effect afterward. The half-maximal concentration for adenosine diphosphate was about 12 muM. Ouabain inhibited synthesis. The ionophore gramicidin had no significant effect on the level of phosphoenzyme nor on the rate nor on the extent of synthesis of adenosine triphosphate. The detergent Lubrol WX reduced the rate of phosphoenzyme break-down and the rate of synthesis but did not affect the final recovery. Phospholipase A treatment inhibited synthesis. In a steady state, the enzyme catalzyed a slow ouabain-sensitive incorporation or inorganic phosphate into adenosine triphosphate. These results and other suggest that binding of sodium ion to a low affinity site on phosphoenzyme formed from inorganic phosphate is sufficient to induce a conformational change in the active center which permits transfer of the phosphate group to adenosine diphosphate.


The mechanism of activation of rabbit plasminogen by urokinase.

The data presented in this paper show that when rabbit plasminogen is activated to plasmin by urokinase at least two peptide bonds are cleaved in the process. Urokinase first cleaves an internal peptide bond in plasminogen, leading to two-chain disulfide-linked plasmin molecule. The plasmin heavy chain of molecular weight 66,000 to 69,000 possesses an NH2-terminal amino acid sequence identical with the original plasminogen (molecular weight 88,000 to 92,000). The plasmin light chain of molecular weight 24,000 to 26,000 is known to be derived from the COOH-terminal portion of plasminogen. The plasmin generated during the activation of plasminogen is capable, by a feedback process, of cleaving a peptide of molecular weight 6,000 to 8,000 from the NH2 terminus of the heavy chain, producing a proteolytically modified heavy chain of molecular weight 58,000 to 62,000. Plasmin also can cleave this same peptide from the original plasminogen, yielding an altered plasminogen of molecular weight 82,000 to 86,000. This plasmin-altered plasminogen and the plasmin heavy chain derived from it by urokinase activation process NH2-terminal amino acid sequences which are identical with each other and with the plasminolytic product of the original plasmin heavy chain. These studies support a mechanism of activation of plasminogen by urokinase which involves loss of a peptide located on the NH2 terminus of plasminogen. However, these same results show that this NH2-terminal peptide need not be released from rabbit plasminogen prior to the cleavage of the internal peptide bond which leads to the two-chain plasmin molecule. Furthermore, these studies show that urokinase cannot remove this peptide from either the original rabbit plasminogen molecule or from the heavy chain of the initial plasmin formed.


Control of pyruvate dehydrogenase activity in intact cardiac mitochondria. Regulation of the inactivation and activation of the dehydrogenase.

The control of pyruvate dehydrogenase activity by inactivation and activation was studied in intact mitochondria isolated from rabbit heart. Pyruvate dehydrogenase could be completely inactivated by incubating mitochondria with ATP, oligomycin, and NaF. This loss in dehydrogenase activity was correlated with the incorporation of 32P from [gamma-32P]ATP into mitochondrial protein(s) and with a decrease in the mitochondrial oxidation of pyruvate. ATP may be supplied exogenously, generated from endogenous ADP during oxidative phosphorylation, or formed from exogenous ADP in carbonyl cyanid p-trifluoromethoxyphenylhydrazone-uncoupled mitochondria. With coupled mitochondria the concentration of added ATP required to half-inactivate the dehydrogenase was 0.24 mM. With uncoupled mitochondria the apparent Km was decreased to 60 muM ATP. Inactivation of pyruvate dehydrogenase by exogenous ATP was sensitive to atractyloside, suggesting that pyruvate dehydrogenase kinase acts internally to the atractyloside-sensitive barrier. The divalent cation ionophore, A23187, enhanced the loss of dehydrogenase activity. Pyruvate dehydrogenase activity is regulated additionally by pyruvate, inorganic phosphate, and ADP. Pyruvate, in the presence of rotenone, strongly inhibited inactivation. This suggests that pyruvate facilitates its own oxidation and that increases in pyruvate dehydrogenase activity by substrate may provide a modulating influence on the utilization of pyruvate via the tricarboxylate cycle. Inorganic phosphate protected the dehydrogenase from inactivation by ATP. ADP added to the incubation mixture together with ATP inhibited the inactivation of pyruvate dehydrogenase. This protection may result from a direct action on pyruvate dehydrogenase kinase, as ADP competes with ATP, and an indirect action, in that ADP competes with ATP for the translocase. It is suggested that the intramitochondrial [ATP]:[ADP] ratio effects the kinase activity directly, whereas the cytosolic [ATP]:[ADP] ratio acts indirectly. Mg2+ enhances the rate of reactivation of the inactivated pyruvate dehydrogenase presumably by accelerating the rate of dephosphorylation of the enzyme. Maximal activation is obtained with the addition of 0.5 mM Mg2+..


Purification and characterization of myosin from the clonal rat glial cell strain C-6.

A myosin was isolated from the clonal rat glial cell strain C-6 and compared with rat skeletal muscle myosin. After cell extracts were subjected to gel filtration chromatography in the presence of KI and magnesium pyrophosphate the C-6 myosin was rapidly purified by a procedure similar to that used for skeletal muscle myosin. The C-6 myosin resembles muscle myosin both physically and enzymatically. It contains heavy chains of 200,000 daltons and two classes of light chains of 17,000 and 19,000 daltons in approximately equal molar ratios. This myosin forms bipolar thick filaments in 0.1 M KCl and binds reversibly to skeletal muscle F-actin, the binding being inhibited by MgATP. Skeletal muscle F-actin stimulates the C-6 myosin adenosine triphosphatase 2- to 3-fold in the presence of KCl and Mg2+. The action activation of muscle myosin ATPase at low ionic strength is 10-fold greater than that of C-6 myosin. Ca2+ and EDTA stimulated the ATPase activities of both enzymes. When assayed in the presence of 0.6 M KCl and 1 mM EDTA the skeletal muscle myocin ATPase demonstrates substrate saturation while the C-6 myosin enzyme activity is stimulated by ATP concentrations above 2.5 mM.


Conversion of the arthrodesed hip to a total hip arthroplasty.

Total hip arthroplasty was done in thirty-three patients who had had previous surgical attempts at arthrodesis of the hip. The arthrodesis had failed in nineteen patients, and the other fourteen patients had fused hips with symptoms in the knee and lower back. Thirty-one of the patients had satisfactory functional ability after total hip arthroplasty after one to three and one-half years' follow-up. The two patients in whom the procedure failed had complicating infections.


Metabolism of peripheral nerve myelin in experimental diabetes.

Previous in vitro studies of the metabolism of the peripheral nerve have been based on incorporation of radioactive precursor into components isolated from whole nerve. In this study we have determined incorporation secifically into myelin components of peripheral nerve by isolating myelin after incubating whole nerves with lipid or protein precursors and by determining the specific activity of the components of that membrane. The effect of diabetes on such incorporation was also studied. In the rat, in vitro incorporation of DL-[1-14C]leucine into protein components of myelin was decreased by 30-88% in diabetic animals as compared to controls. The major polypeptide constituent of rat sciatic nerve myelin (mol st 28,000; 58.5% of total mass of proteins) was not labeled in either the diabetic or the control group. In diabetes incorporation rate into a polypeptide of mol wt 23,000, which constitutes 21% of total mass, was approximately one half that of controls. In polypeptides of mol wt 38,000-49,000, which are heavily labeled in normal animals, but constitute only about 5% of total mass of proteins, depression of incorporation was e-en more marked in the diabetics. While these marked differences in incorporation between diabetic and control animals were observed, the amount of protein and its distribution among the constituent polypeptides was the same in both groups. In young rats made diabetic with streptozotocin and young rabbits made diabetic with alloxan, there was a lower rate of incorporation of the lipid precursors, [1-14C]sodium acetate or [3H]water, into myelin components. In older animals of both species incorporation in the controls was considerably lower than in the yount animals, and the effect of diabetes was no longer apparent. In nondiabetic animals, the in vitro addition of insulin (10-7 M) stimulated incorporation of DL-[1-14C]leucine into myelin proteins 1.6-3.1 times that of controls. This stimulation by insulin in vitro was not seen in diabetic animals. In animals in which diabetes had spontaneously recovered, however, incorporation rate in the in vitro experiments approached that of controls and were significantly above that in animals whose diabetes persisted. Since myelin is the palsma membrane of the Schwann cell, these studies provide evidence that the Schwann cell is affected by insulin and that some aspects of the metabolism of myelin are altered in insulin-deficient states.


Visual hallucinations and delirium during treatment with amantadine (Symmetrel).

In 13 elderly patients, 12 of whom had Parkinson's disease, visual hallucinations and delirium developed as a side effect of amantadine hydrochloride (Symmetrel) therapy. The symptoms promptly disappearred when amantadine was discontinued. Thereafter, each parkinsonian patient was treated satisfactorily with levo-dopa. Treatment with a combination of amantadine and an anticholinergic agent increases the likelihood of delirium because of the hazard of retention of urine. Although amantadine is effective in the treatment of Parkinson's disease in the elderly, the incidence of delirium as a complication seems higher in this age group.


Cytotoxic effects of antigen- and mitogen-induced T cells on various targets.

Mitogen-induced cytotoxicity was studied by culturing mouse spleen cells with an optimum mitogenic dose of concanavalin A (Con A) for 2 days and, after washing, assessing their ability to lyse tumor targets. We show that rapid lysis occurs only when the correct concentration of an agglutinating mitogen (phytohemagglutinin P (PHA) or Con A) is present in the assay. PHA is more efficient than Con A in revealing the cytotoxic effect. The Con A-induced effector cytotoxic cell is shown to be sensitive to anti-theta serum and complement. Cytotoxic T cells were also induced by H-2 different allo-immunization. These effector cells specifically lyse targets which bear the immunizing H-2 antigens in the absence of PHA. When PHA is present in the assay, they will also lyse syngeneic tumor targets nonspecifically. Since not all dividing T cells (e.g., T lymphomas, and T blasts induced by M locus different, H-2 similar mixed lymphocyte culture) lyse PHA-P815, we propose that this assay measures only a subset of effector T cells, namely, cytotoxic T cells, regardless of their antigen specificity. The tumor cells, P815 and EL4, are sensitive both to antigen-specific T cell-mediated lysis in the absence of PHA and to nonspecific, PHA-revealed lysis. Small lymphocytes, T cell blasts, and B cell blasts are sensitive to lysis by T cells which are directed against H-2 antigens on their surface, but are not very susceptible to nonspecific T cell lysis in the presence of PHA. The reason for this difference in susceptibility of tumor and normal cells to PHA-revealed nonspecific T cell lysis is not known but may have some relevance to in vivo tumor rejection.


Characterization of immune response and mixed lymphocyte reactions in selected intra-H-2 recombinant strains.

The genes controlling the immune response to the random linear terpolymers GAT and GLpro have been mapped in the Ir-1A or Ir-1B subregions with five lines of H-2-s/H-2-a recombinant mice. The I region also contains a third subregion, I-C, which codes for a lymphocyte-activating determinant, Lad2, controlling mixed lymphocyte reactivity. The mixed lymphocyte responses associated with I-C region disparity are compared with the stimulation noted with other H-2 region differences. Unidirectional lymphocyte activitation was associated with the Lad2 determinants in selected combinations.


Genetic control of responses to bacterial lipopolysaccharides in mice. II. A gene that influences a membrane component involved in the activation of bone marrow-derived lymphocytes by lipipolysaccharides.

C3H/HeJ mice contain a defect in a single autosomal locus which is not linked to the H-2 histocompatibility or the heavy chain allotype loci that restrict immune, mitogenic, and polyclonal responses to bacterial lipopolysaccharides (LPS). Adult thymectomized C3H/HeJ mice that have been irradiated and reconstituted with C3HeB/FeJ bone marrow cells respond well to LPS. Cell-mixing experiments using C3H/HEJ-C3HeB/FeJ spleen cultures show that the failure of C3H/HeJ spleen cells to support responses to LPS is not due to nonspecific or LPS-induced suppressive events, or the lack of accessory cell types. C3H/HeJ and C3HeB/FeJ spleen cells bind LPS and respond to other B cell mitogens equally well. We suggest that the B lymphocytes of C3H/HeJ mice have a defect in a membrane component that is activated via interaction with LPS, and initiates the intracellular events that lead to cell proliferation.


Macrophage activation of allogeneic lymphocyte proliferation in the guinea pig mixed leukocyte culture.

The role of the macrophage in the guinea pig mixed leukocyte culture was investigated. Macrophages obtained from oil-induced peritoneal exudates, peritoneal wash-out cells, spleen, and alveolar washings were found to be effective stimulators of allogeneic lymph node and splenic lymphocyte DNA synthesis. The stimulatory properties of macrophages proved radioresistant but viability dependent. Unfractionated lymph node cells or adherence column purified lymph node lymphocytes and thymocytes were only minimally active as stimulators, even in the presence of macrophages syngeneic to the responder lymphocytes. Allogeneic fibroblasts, polymorphonuclear leukocytes, L2C leukemia cells, and xenogeneic (murine) macrophages failed to simulate. These data provide evidence that the macrophage is the predominant stimulator of the mixed leukocyte culture in the guinea pig.


Suppressor cells in tumor-bearing mice capable of nonspecific blocking of in vitro immunization against transplant antigens.

Spleen cells from mice with progressively-growing methyl-cholanthrene-induced tumors, when immunized in vitro against transplant alloantigens, developed less cytotoxic activity against these antigens as measured by a short-term chromium-release assay than did spleen cells from normal mice. The hyporesponsiveness of spleen cells from the tumor-bearing mice seemed to be due to the presence of suppressor cells which could be removed by nylon-column passage but not by anti-theta treatment and which, in mixture experiments, could inhibit the response of normal spleen cells. The suppression appeared to occur at the sensitization stage and not at the effector stage of the in vitro tests. No evidence was found for mediation of the suppression by soluble factors. These observations emphasize the growing importance of suppressive mechanisms in tumor immune systems.


Demonstration of immune complexes in spontaneous and histamine-induced lesions and in normal skin of patients with leukocytoclastic angitis.

Clinical and laboratory observations have strongly suggested that leukocytoclastic angitis is an immune complex disease. Since immune complexes can be visualized as electron-dense deposits by electron microscopy (EM), this method was used in conjunction with direct immunofluorescence (IF) to determine whether complexes could be demonstrated in spontaneous lesions, and in uninvolved skin in which the vessels were made permeable by the local injection of histamine. Histamine-induced wheals were produced in the uninvolved skin of patients with active angitis. In the resulting wheal, EM studies revealed electron-dense deposits characteristic of immune complexes in postcapillary venules and direct IF studies demonstrated complement and immunoglobulins in the vessel walls. Neutrophils in varying stages of disintegration were present thereby reproducing the histopathologic changes of spontaneous lesions. EM and IF studies of nonmanipulated uninvolved skin also revealed electron-dense deposits and immune reactants in the vessel walls. Neutrophils were not present, however. This observation indicates that immune complexes are deposited in vessels before tissue damage ensues. Study of spontaneous lesions older than 24 hr revealed only fibrin by EM and no immune reactants by direct IF. In spontaneous lesions less than 24 hr old, electron-dense deposits and fibrin were seen by EM, and complement and immunoglobulins by IF. Histamine-induced wheals should be a useful device to investigate patients with disorders that have an immune complex pathogenesis.


The reactivity of derivatives of digoxin and digitoxin as measured by the Na-K-atpase displacement assay and by radioimmunoassay.

Derivatives of digoxin and digitoxin were measured by the Na-K-ATPase displacement assay and by radioimmunoassay and the data compared with biological potency of these compounds. There was a slightly higher affinity of digoxigenin-bis-digitoxoside as compared with digoxin using digoxin-specific antiserum and considerably less affinity of digoxigenin and dihydrodigoxin than the parent compound in the same system. A similar trend was observed for the derivatives of digitoxin using digoxin-specific antiserum. The recovery on extraction of some of the derivatives of digoxin and digitoxin differed from that of the parent compounds in the ATPase assay. The potency of the derivatives of these drugs in displacing 3H ouabain also differed from the parent compoundmboth the recovery on extraction and the potency for displacing ouabain must be considered in the estimation of the contribution of the derivatives of digoxin or digitoxin to the result of Na-K-ATPase assay. Quantitative information of the metabolites of digoxin and digitoxin in normal and pathological conditions is needed to properly interpret assay data obtained either by radioimmunoassay or by ATPase assay.


Effect of ethanol ingestion on duodenal calcium transport.

Effects of chronic ethanol ingestion on duodenal calcium transport were investigated in rats ingesting 20 per cent ethanol. Calcium transport was inhibited by ethanol ingestion and the defect could not be reversed by vitamin D or 25-hydroxycholecalciferol administrationmethanol ingestion by vitamin D-deficient rats did not further suppress transport activity nor interfere with an increase in transport induced by vitamim D. Levels of intestinal calcium-binding activity were not suppressed. Brush-border alkaline phosphatase activity was suppressed by chronic ethanol ingestion as compared to ad libitum-fed control animals and administration of vitamin D to animals ingesting ethanol restored alkaline phosphatase activitymthe results suggest that ethanol interferes with calcium transport by a mechanism at least in part independent of the vitamin D pathway and that changes in alkaline phosphatase and calcium transport, although both affected by vitamin D, may represent independent metabolic consequences.