Establishment and characterization of cell lines from Gross virus-induced rat leukemia.

Cells from thymus and/or bone marrow tissues of either normal or Gross virus-infected W/Fu rats were cultivated individually or mixed. Four continuous cell lines were obtained: 1) individual cultivation of the cells from thymic lymphoma, 2) combined cultivation of cells from thymic lymphoma and normal bone marrow, 3) bone marrow of a leukemic rat and normal thymus, and 4) bone marrow of a leukemic rat supplemented with cell-free extracts of normal thymus homogenates. The morphological features were similar in these 4 cell lines. The cell correspond morphologically to lymphoblasts, grow rapidly in suspension, and are transplantable in newborn rats. The cells release abundant Type-C virus particles. The inoculation of cell-free culture fluid into newborn rats resulted, after 7-9 weeks, in the development of thymic lymphoma in high incidence.


Temporary cell cycle arrest in neural and extraneural developing rat tissues after exposure to methyl--and ethylnitrosourea.

8 days old rats were exposed to 20 or 100 mg/kg b.w. of either Methylnitrosourea (MNU) or Ethylnitrosourea (ENU), followed by injection of 10 muCi/g b.w. of (3H-methyl)-Thymidine. After a 100 mg dose of MNU or ENU in both neural and extraneural tissues a total inhibition of S-phase radioactivity is observed that lasts longer for MNU than for ENU. Moreover reappearance of S-phase cells in the neural tissues is later (36-48 h) than in the extraneural tissues (24-36 h) for both drugs. In both neural and extraneural tissues reappearance of S-phase cells is consistently found to occur about 12 h earlier than recurrence of M-phase cells. After a 20 mg dose of MNU or ENU in both neural and extraneural tissues a clear decrease in S-phase radioactivity is found after a 6 h' interval only. There are only slight differences between the various tissues and drugs. In developing rat tissues there is obviously a trend for both mitotic activity and S-phase radioactivity to decrease with increasing single doses of MNU or ENU. Our results point to an arrest in or before entering the S-phase of the cells involved. The more pronounced cytotoxic activity of MNU as compared to ENU is discussed. Recurrence of DNA synthesis and re-entrance of damaged cells into their cycle prior to the elimination of altered bases from DNA might be of importance for the problem of oncogenesis.


Endodermal sinus tumour of the ovary: a comparative light and electron microscopic study.

An endodermal sinus tumour of the ovary from a 12 year old girl is analysed light- and electron microscopically. The histological appearance is characterized by glandular-papillary structures and microcystic-reticular areas with inclusion of occasional endodermal sinuses. By electron microscopy immature and highly differentiated cells can be distinguished. In analogy to certain structures in the human yolk sac the most differentiated cells in the tumour are regarded as neoplastic endoderm. On the basis of transitional forms between immature and differentiated cells a development of the latter from the former is suggested. The cells in mesenchyme-like areas differ from those of solid and glandular parts merely in the degree of cytoplasmic differentiation but are otherwise believed to represent the same "cell line". In addition to other possible cell functions the cytoplasmic features of the differentiated cell indicates a protein synthesis and secretion. From our observations it is concluded that the endodermal sinus tumour originates from germ cells and differentiates into yolk sac endoderm. The ultrastructural differences between this tumour and clear cell carcinomas of the ovary are discussed.


The effects of some carcinogenic nitroso compounds on the rat liver nucleolus.

The hepatocyte nucleoli of rats undergo a variety of specific and nonspecific alterations after the administration of 8 nitroso compounds and 5 corresponding non-nitroso compounds. After the oral administration of dimethylnitrosamine and diethylnitrosamine, and unusual segregation of the nucleolus was encountered in the cells with karyorrhexis. Most characteristic of the segregation are the scattered ribosome-like granules at the outermost layer of the altered nucleolus. Frequently microspherules with halos 150 nm in diameter are found in all rats treated with nitroso compounds, which have been reported to be carcinogenic in the organs other than the liver. With the exception of butylurea and butylamine no such specific microspherules were found in the nucleoli after the administration of corresponding non-nitroso compounds.


[Ultrastructure and formal pathogenesis of embryonal rhabdomyosarcoma (author's transl)].

An embryonal rhabdomyosarcoma of the nasopharynx of a 10 year old boy is analysed with light and electron microscopy. With regard to cell shape and cytoplasmic features the following four tumour cell types could be distinguished: 1. Undifferentiated mesenchymal cells with a big loosely packed nucleus and a small cytoplasmic rim with only few cell organelles; 2. Undifferentiated tumour cells with a broad cytoplasmic body which contains a dense network of nonspecific intermediate filaments with a diameter of about 100 A; 3. Immature rhabdomyoblasts with randomly orientated specific myofilaments; 4. Fully differentiated rhabdomyoblasts with well developed myofibrils often showing a sarcomeric pattern. Glycogen deposits which were seen in great masses in many tumour cells were regarded to result from degenerative processes within the tumor. The cellular stages in the development of rhabdomyoblasts are basically identical to those known from the embryogenesis and regeneration of striated muscle. From these observations the two following developmental pathways are suggested: 1. Origin of the tumour from an undifferentiated mesenchymal cell; 2. Atypical regeneration of striated muscle which terminates in malignant progressive tumour growth. At present, the body of information about rhabdomyosarcomas supports the assumption of an origin from immature mesenchymal cells. Nevertheless, the second theory cannot be totally excluded.


Transplacental effects of nitrosamines in Syrian hamsters: I. Dibutylnitrosamine and nitrosohexamethyleneimine.

The transplacental carcinogenic effects of dibutylnitrosamine (DBN) and nitrosohexamethyleneimine (N-6-MI) were examined in Syrian hamsters. A proportion of both substances reached the fetal tissue unaltered. No macroscopic malformations were observed in the offspring; however, postnatal mortality was high. Respiratory tumours were found upon histologic examination of surviving animals. Single doses of 30 mg/kg body weight (b.w.) DBN and 2 doses of 10 mg/kg b.w. N-6-MI did not induce tumours in the P-generation, but led to a low tumour incidence in the F1-generation (DBN, 7.0%, N-6-MI, 2.0%). Treatment for up to eight days during the second half of pregnancy led to a higher tumour incidence in the P-generation (DBN, 22%; N-6-MI, 20%), than in the F1-generation (DBN, 6.0%; N-6-MI, 10%).


Life-span investigations for carcinogenicity of some immune-stimulating, immunodepressive and neurotropic substances in Sprague-Dawley-rats.

In a long-term trial in a total of 858 Sprague-Dawley-rats of both sexes the possible carcinogenicity of 12 substances was investigated. Complete Freunds' adjuvant which is among the immune-stimulating substances was found to decrease significantly the lifetime of the animals; whereas an increase in tumor incidences was not observed. Bacille Calmette-Guérin (BCG), human albumin, and vitamin-A-acid were neither carcinogenic, nor had they an influence on the survival times. Rabbit-antirat lymphocytic serum which is a well-known immunodepressive agent caused a significant decrease in survival times, it was not found to be carcinogenic. The known carcinogenic properties of cyclophosphamide were verified. The applied dose of this compound significantly reduced the mean survival times of the animals. Hydrocortisone and amethopterin were found to have no carcinogenic effects. Atropine, pilocarpine, nicotine, and phenyl-ethyl-barbituric acid (phenobarbital) which are known to have neurotropic effects were tested. Nicotine and phenobarbital were found to diminish the mean survival times. None of these neurotropic substances was found to be carcinogenic.


Experimental investigations on the influence upon the chemical carcinogenesis. IInd communication: studies with 3,4-benzopyrene.

After subcutaneous application of 0.5 mg 3,4-benzopyrene (BP) to Sprague-Dawley-rats on the 2nd day of life, 50% of the animals developed local fibrosarcomas after 250 +/- 70 days. Additional treatment with immunostimulating (BCG, albumin, vitamin A-acid) or immunodepressive agents (hydrocortisone, cyclophosphamide, methotrexat) which was started 6 days after birth and maintained throughout life, did not influence carcinogenesis with respect to tumor incidences and induction periods of tumors.


Experimental investigations on the influence upon the chemical carcinogenesis. IIIrd communication: studies with 1,2-dimethylhydrazine.

After ten times monthly, subcutaneous, injections of 30 mg/kg 1,2-dimethylhydrazine (DMH) to Sprague-Dawley-rats, malignant tumors were found in approximately 90% of the animals after a mean induction period of approximately 330 days. Injections were started on the 2nd day of life and maintained during 10 months. 72% of the tumors induced were formed in the colon, 17% were squamous cell carcinomas of the ear duct, adenosarcomas of the kidney and hepatocellular carcinomas were found in 13 and 11% of the animals, respectively. Additional treatment with immunodepressive (cyclophosphamide, methotrexat, hydrocortisone) and immunostimulating substances (BCG, albumin, vitamin A-acid) as well as an enzyme-stimulating agent (Luminal) did not alter incidences and induction periods of tumors. After application of a vegetarian diet, the rate of liver and kidney tumors was diminished significantly and induction periods of intestinal and ear duct tumors increased.


Carcinogenic effects of different nitroso-compounds in Chinese hamsters. II. N-nitrosomorpholine and N-nitrosopiperidine.

A total of 320 Chinese hamsters (Cricetulus griseus) (CH) were subcutaneously (s.c.) treated with 1/5, 1/10, or 1/20 LD50 of N-nitrosomorpholine (NM) or N-nitrosopiperidine (NP). In the CH, NM and NP both produced up to a 100% rate of papillomas of the cheek pouch, tongue, pharynx, esophagus and forestomach. Occasionally squamous cell carcinomas also developed in these organs. A high rate of hepatomas was realized by NP.


Synthesis of purine and pyrimidine substituted nitroxides.

Reaction of 6-hydrazinopurine and 2-hydrazinopyrimidine with 2, 2, 6, 6-tetramethylpiperidone-1-oxyl gives 2, 2, 6, 6-tetramethylpiperidone-1-oxyl-(6-purinyl) hydrazone (3b) and 2, 2, 6, 6-tetramethylpiperidone-1-oxyl-(2-pyrimidinyl) hydrazone (4b) respectively. Compound 3b is inactive even at high dose levels against TLX/5 lymphoma and 3b and 4b are both inactive in tests using the L1210 leukaemia system.


[Redundant nucleotide-sequences in DNA from wheat (author's transl)].

DNA from dormant and germinated embryos exhibit differences in the kinetics of reassociation of thermal denatured nucleic acid. Within 24 h of germination a reduction of the renaturation, expressed as Cot1/2 of the redundant fraction, is detectable.


A cell for simultaneous measurements of optical and electrical properties of black lipid membranes.

A universal cell was constructed, which allows measurements of the resistance (d.c.), capacity, optical reflectivity, and spectral absorption of black lipid films simultaneously. These measurements reflect different properties of the films, supplementary to each other. The purpose is to eliminate differences of experimental conditions especially in recombination experiments with proteins. The cell is totally symmetric, can be completely disassembled, and the parts are exchangeable. It allows application of several methods for film production, and is held together simply by mechanical pressure, without need of any sealing agent. The inner parts are exclusively of glass and teflon. The fluid volume of the cell is 7-15 ml and the insulation between the two cell compartments is better than 10(14) Ohms.


Studies on phenylpyruvic acid. I. Keto-enol tautomerism.

The tautomerism of phenylpyruvic acid (PPA) and its sodium salt was investigated. The 1H and 13C spectra of PPA in aprotic solvents and in methanol show almost complete prevalence of the enol form, whereas the keto form prevails only in water. The Z configuration was assigned to the sole enol tautomer present on the basis of the value of the vicinal coupling constant [3J1H, 13COOH]=3.7 Hz. A small amount of the hydrated form of PPA (1-phenyl-2, 2-dihydroxylpropanoic acid) was found in the aqueous solution of its sodium salt and in buffer solution (pD=6) of PPA. By means of infrared spectroscopy one can conclude that crystalline PPA is in the enol whereas its sodium salt is in the keto form. The keto-acid was not obtained in the solid state. The colorimetric method for testing PPA traces in urine depends on the formation of a enol-Fe3+ complex (2:1) which appears stable in dimethylsulfoxide (DMSO).


Correlation of the photosynthetic reduction of p-(diazonium-) benzenesulfonic acid with the increased binding of the probe to the thylakoid membrane.

The reactions of chloroplast thylakoid lamellae with the chemical probe p-(diazonium-) benzenesulfonic acid (DABS) in the light have been reinvestigated. In contrast to a previous report, electron transport from a photosystem I electron donor to methylviologen was found to be inhibited by this treatment. During the incubation of chloroplasts with DABS in the light, the probe is altered with high rates. Under aerobic conditions, a concomitant oxygen uptake is observed, which is stoichiometric to the amount of DABS altered. Under anaerobic conditions, the binding of the 35S-labeled probe to the membranes in the light is stimulated 2-3 fold as compared to the binding under aerobic conditions. The data are taken as evidence that the photoreduction of the probe rather than a conformational change of the membrane may be at least partially responsible for the increased reagent binding observed in the light.


Enzymic capacities for chlorophyll biosynthesis. Activation and de novo synthesis of enzymes.

A previously published working model for the regulation of chlorophyll formation has been tested studying early steps of chlorophyll and porphyrin biosynthesis in developing cotyledons of Helianthus annuus. The activities of delta-aminolevulinate synthetase (ALAS), delta-aminolevulinate dehydratase (ALAD), and the porphobilinogenase complex (PBGase) at any given time have been found to be strongly associated with endogenous developmental processes. Highest activities in darkness have been observed at times when maximum chlorophyll formation would have occurred had the plants been exposed to light. Only in the case of ALAS was the maximum activity in light much greater than that observed in the dark. Density labeling experiments and other data suggest that enzyme synthesis is mediated both by development and by illumination. Moreover, ALAS activity appears to be subject to inhibition, presumably by products of the porphyrin biosynthesis, as indicated by halflife experiments. Rapid enzyme degradation in the absence of light seems to be less probable. Slight ALAS activity in darkness is present as long as the plastids are not fully developed. In contrast to findings with cell cultures of tobacco, in Helianthus cotyledons ALAS certainly plays the main role in the regulation of chlorophyll biosynthesis. Nevertheless, increasing activities of the succeeding enzymes, located in the plastids, ensure that increased concentrations of delta-aminolevulinate (ALA) are drawn into the chlorophyll biosynthetic pathway. The experiments corroborate the suggestion that chlorophyll biosynthesis is controlled by different but interdependent mechanisms. The dominant regulatory mechanism is dependent on the stage of development.


The effect of deoxycholate-treatment to the photoreactions of the active pigments in photosynthesis.

In the heavy fraction of deoxycholate-treated spinach chloroplasts the chlorophyll aII activity is high and the chlorophyll aI activity is low when no artificial electron donor is added. The addition of the photosystem I donor system N-methyl-phenazonium sulphate plus sodium ascorbate (PMS + Asc) leads to a complete reactivation of the chlorophyll aI reaction. The addition of the photosystem II donor system p-benzohydroquinone plus sodium ascorbate (HQ + Asc) leads to an inhibition of the chlorophyll aII activity. From these results we conclude: 1. Besides an interruption of the linear electron flow between the two photosystems deoycholate-treatment leads to a block of the electron flow from water to photosystem II. 2. In deoxycholate-treated chloroplasts the linear electron flow in photosystem II just like in Tris-washed, heat-treated or aged chloroplasts, is replaced by a cyclic one.


The chlorophyll aII reaction in trypsin-treated spinach chloroplasts in the presence of potassium ferricyanide.

In trypsin-treated spinach chloroplasts there is no linear electron flow from water to potassium ferricyanide. The chlorophyll aII reaction, however, is still active but insensitive to 3-(3, 4-dichlorophenyl)-1, 1-dimethylurea. From this we conclude that ferricyanide and trypsin together stimulate a mini-cycle in photosystem II.


Ultrastructural changes in in vitro ageing spinach chloroplasts.

Ultrastructural changes in in vitro ageing spinach chloroplasts have been studied in detail. Prolonged storage caused swelling of the chloroplasts due to the increase in the thickness and spacing of the thylakoid membranes. The increase in the thickness of the membrane is partly accompanied by the release of lipids. Addition of crystalline bovine serum albumin was found to stabilize the membrane structures. Storage of the chloroplasts at 77 degrees K even though it resulted in complete breakage of the whole chloroplasts, maintained the thylakoid structures in a highly intact form.


[Size comparison of poly (A)-RNA from polysomes and from nuclei of the yeast Saccharomyces cerevisiae (author's transl)].

The size of poly (A)-RNA from polysomes and cell nuclei of the yeast Saccharomyces cerevisiae was investigated. Pulslabelled cells ([14C] adenin) were cracked by the French press; polysomes and nuclei were separately isolated and the RNA was finally extracted with phenol. The separation of poly (A)-containing and poly (A)-lacking fractions was achieved by oligo (dT) cellulose. These fractions were characterized by sedimentation analysis. The main portion of polysomal poly (A)-RNA sedimented with a rate of 8 to 14S, whereas the poly (A)-RNA of nuclei exhibited a sedimentation rate of 12 to 17S. Thus nuclear poly (A)-RNA is about 20-30% larger than polysomal poly (A)-RNA.


[Chromosomal structures of Pseudomonas testosteroni. I. Isolation and characterization of the chromosomal complexes. (author's transl)].

After lysis of Pseydomonas testosteroni with lysozyme and non-ionic detergents different DNA-protein complexes can be separated in 5-25% (w/v) neutral sucrose gradient. The protein to DNA ratio of these complexes varies between 0.5-4.5 to 1, whereby the faster sedimenting forms contain more protein than the slower sedimenting ones. Different initial rates of DNase digestion may indicate various degrees of DNA packing in these complexes. The chromosomal complexes of Pseudomonas testosteroni are relatively stable towards pronase. Treatment with RNase or sodium dodecylsulphate is accompanied by a dramatic increase in viscosity and decrease in relative density. It suggests that DNA in these complexes is maintained in its supercoiled form by RNA molecule(s) in a similar way as in isolated chromosome of E. coli.


Variations in melanin and riboflavin content of amphibian liver under the influence of reserpine and amphetamine.

By means of paper chromatography, Lactobacillus casei test and the staining method with the Feulgen reaction, the effects of reserpine and amphetamine on riboflavin and melanin were studied in the liver of Triturus cristatus. It was demonstrated that the concentrations of both melanin and riboflavin can be altered by the influence of these drugs. From the results obtained it is suggested that riboflavin shows a correlation with the density of melanin.


[Pathobiology of carcinogenesis and the growth of neoplasms].

The importance of carcinogenicity by chemical substances is reviewed with regard to the increasing number of human neoplasms (1955: 39703; 1972: 62556) in the German Democratic Republic. The organotropic action of some carcinogens, the transplacental tumour induction, the endogenous formation of oncogenic N-nitroso compounds from inactive precursors in the stomach of experimental animals, and the role of carcinogens derived from plants are analysed. Up to now it remains uncertain, whether DNA and RNA viruses responsible for tumour development in different species of animals may effect tumour growth in human beings as well.


[Histochemistry of the pulmonary alveolar proteinosis. A contribution to the formal pathogenesis (author's transl)].

By means of histochemical methods the authors investigated 3 cases of the diffuse and 2 cases of the focal form of pulmonary alveolar proteinosis (2 women and 3 men, aged from 25 to 58 years). In the main the results of other investigators concerning distribution and occurrence of proteins and lipids are confirmed. In contrast to the results of other investigators, the authors detected acid mucopolysaccharides also in the granules of the intraal veolar substance. Presumable the injuring substance causes proliferation, desquamation, and breakdown of the alveolar cells. In this way both the granules and clots are formed, the latter ones are decomposed in steps of granules. Obviously parts of the blood are mixed with the granular part of the intraalveolar substance.


Hormonal changes during the menstrual cycle of the baboon (Papio hamadryas).

Eleven unconjugated steroids were measured daily during a complete cycle in the peripheral plasma of 6 normally menstruating baboons (Papio hamadryas) by means of a radioimmunoassay procedure and the levels were compared with those found previously in 17 normally menstruating women. The patterns of progesterone and 20alpha-dihydroprogesterone were very similar to those found in women throughout the entire menstrual cycle. However, the ratio of these steroids differed markedly from that found in women. A great similarity of the follicular phase and the peri-ovulatory period profiles was observed in both species for 17-hydroxy-progesterone, pregnenolone, androstenedione, oestradiol and oestrone. The pattern of oestradiol:oestrone ratios was similar in both species. However, an elevation of the above five steroids, typical for the human luteal phase, was not found in baboons. The increase of testosterone values, seen in women at mid-cycle, was not detected in baboons. The plasma concentrations were lower in baboons than in humans for all the above steroids.


Histochemical and biochemical research on the pallial gland of Lithophaga (Lithodomus) lithophaga L.

Histomorphological staining demonstrated 2 types of cells in the pallial gland of the mantle of Lithodomus lithophaga: gland cells containing glycolipoprotein and interstitial cells, containing lipid droplets. Biochemical analysis aiming at separate and evaluate lipidic and peptidic components were carried out by lipid extraction, T.L.C. separation and colorimetric assays. Phospho- and sulfolipids were separated from non-polar lipids. Sialo-cerebrosides (gangliosides) are missing. Hydrophobic peptide component was extracted and aminoacid analysis was made. The presence of acid (phosphate) group in the gland secretion is discussed in relation to the function of the secretion (Ca uptake and rock boring). The prominent glycogen storage is considered as an important energy source to be used for osmotic work and to synthesize glycosidic stones for glycolipo-protein secretion.


Evaluation of the pain suppressive effect of different frequencies of peripheral electrical stimulation in chronic pain conditions.

The pain suppressive effect of low (2/s) and high (50-100/s) frequency electrical stimulation was studied in 12 patients with severe chronic pain in the back and/or the legs. All patients were subjected to a thorough physical examination before and 2-3 weeks after a series of stimulation sessions. The stimulation was applied via surface electrodes to areas both segmentally related and segmentally unrelated to the regions of chronic pain. Low frequency stimulation induced a partial pain relief in only one patient whereas stimulation with high frequency gave a suppression of pain in seven patients. The effect was short-lasting in most cases and the pain started to increase usually within 30 min. It is concluded  that the observed pain suppression is not due to psychological factors but to more basic neurophysiological mechanisms. Augmentation of the effect after repeated stimulation sessions was not observed, neither was there any alteration in the neuro-orthopedic status nor any lasting pain relief in any patient.


Morphological and biochemical studies of a case of mucopolysaccharidosis II (Hunter's syndrome).

An autopsy case of a 19-year-old boy who had shown typical gargoyle features, strictly consistent with mucopolysaccharidosis type II (Hunter's syndrome) was reported. Histologically, cytoplasmic vacuolar change was found in hepatocytes, sinusoidal epithelium of spleen, follicular cells of thyroid, Sertoli cells of testis, chromophobe cell of pituitary and generalized fibroblast-like cells including meninges, cardiac valve and periosteum. The vacuoles consisting of membrane-bound structures with flocculus protein-like material and occasional electron dense bodies on electron microscopy, were considered to be the site of mucopolysaccharide deposition by histochemical analysis. Deposition of lipid material consistent with so-called membranous cytoplasmic body was observed in the neurons of central, peripheral and autonomic nervous system. Hepatosplenomegaly could be explained by cytoplasmic deposition, but the cause of cardiomegaly remained further to be studied. Biochemically hepatic mucopolysaccharide was identified as heparan sulfate, while in the kidney dermatan sulfate and heparan sulfate were detected. The correlation between morphology and biochemistry, and between deposition and degeneration was discussed.


Cytological characterization and histogenesis of Ewing's sarcoma.

In addition to a light-microscopical and histochemical investigation of primary and metastic lesions in 27 cases of Ewing's sarcoma, biopsy materials from the primary bone lesions of 7 patients with this neoplasm were examined histochemically, enzyme-cytochemically and electron microscopically to elucidate the histogenesis and nature of the neoplasm. Ultrastructural observation has revealed that besides intracytoplasmic and extracellular deposition of glycogen the tumor cells possess several cytological features characterized by intracytoplasmic microfilaments of varying thickness up to 80 A, occasional appearance of dense patches, fat droplets, desmosomal connections and reminiscent attachment bodies. The tumor cells are mostly round, oval or polygonal in shape, but spindle or elongated cells are intermingled and occasionally contain well-developed rough endoplasmic reticulum, resembling pericytes or fibroblasts. In the intercellular spaces amongst the tumor cells, varying amounts of variable-shaped amorphous materials are found, which are compatible with acid mucopolysaccharides and glycoproteins histochemically verified. These findings may suggest that Ewing's sarcoma is a highly malignant neoplasm originating from a transitional cell developed from pericytes to vascular smooth muscle cells in the bone marrow.


The myocardial capillary vasculature in exercising animals with increased cardiac pressure load.

The myocardial capillary reaction was studied in normal rats subjected to swimming exercise and in rats in which aortic stenosis had been produced at various time intervals before the swimming exercise was commenced. The data obtained indicated that neoformation of myocardial capillaries during swimming exercise occurred only in rats without aortic stenosis. It is concluded that a heart subjected to an increased pressure load is less able to respond to a superimposed volume load by an increase in its capillary supply than is a normal heart.


The influence of guanoxane on the submaxillary glands of rats. Another model of neurogenic sialoses.

In extracts of submaxillary glands of rats, treated with the guanethidine derivative, guanoxane (Envacar), alpha-amylase activity and protein concentration were measured. Both parameters were also determined for the alpha-isoamylase patterns obtained following electrophoretic separation of gland extracts in polyacrylamide gels. During the first week of drug application, amylase was found to be congested in the glands. Compared with untreated animals, total amylase concentration and amylase activity per unit protein (specific enzyme activity) were significantly increased in extracts and separated isoenzymes, whereas protein concentration remained unaltered. The described changes reverted to normal after continued drug application for 3 weeks. The possibility is suggested that changes in alpha-isoamylase patterns might eventually be employed for clinical characterization of different types of neurogenic sialosis.


The nature of the training response; peripheral and central adaptations of one-legged exercise.

13 male subjects were studied and placed in 3 groups. Each group exercised one leg with sprint (S), or endurance (E) training and the other leg oppositely or not at all (NT). Oxygen uptake (Vo2), heart rate and blood lactate were measured for each leg separately and for both legs together during submaximal and maximal bicycle work before and after 4 weeks of training with 4-5 sessions per week. Muscle samples were obtained from the quadriceps muscle and assayed for succinate dehydrogenase (SDH) activity, and stained for myofibrillar ATPase. In addition, eight of the subjects performed after the training two-legged exercise at 70% Vo2 max for one hour. The measurements included muscle glycogen and lactate concentrations of the two legs as well as the blood flow and the a-v difference for O2, glucose and lactate.


Identification of the mucopolysaccharides in catecholamine-containing subcellular particel fractions from various rat, cat and ox tissues.

Subcellular fractions containing adrenergic vesicles were obtained from bovine adrenal medulla and splenic nerve, rat and cat heart, and rat spleen. The fractions were washed free from lipids, digested with papain and the mucopolysaccharides (MPSs) then precipitated  from the supernatant with ethanolic potassium acetate. The mpsswere identified by several different methods--microelectrophoresis on cellulose acetate in various media, cellulose column chromatography using elution solvents of increasing ionic strength, and treatment with specific enzymes followed by electrophoresis or column chromatography. The MPS precipitate from all the sources investigated contained dermatan sulphate or a dermatan sulphate-chondroitin sulphate hybrid. in addition, the precipitate from rat spleen was found to contain chondroitin sulphate. Heparan sulphate was found in the precipitates from rat heart and spleen and hyaluronic acid in that from bovine splenic nerve. The finding of sulphomucopolysaccharides in the adrenergic vesicles, probably in a complex with protein, raises the question of the functional significance of such complexes. They might, by analogy with the ion-exchange function of the heparin-protein complex in mast cell granules, play a role in the storage and release of the amines.


In vitro studies on target cells of oncogenic adenoviruses in hamster brain. II. In vitro transformation of brain cells of hamsters at various ages by human adenovirus type 12.

In vitro transformations of brain cells of hamsters of various ages were examined after the administration of human adenovirus type 12 (Ad 12) to determine the type and origin of the target cell. Hamster brain cells at all examined ages were transformed by Ad12. Although the virus was not isolated, virus specific tumor antigen was demonstrated in the transformed cells. The histological features of tumors that developed by transplantation of transformed cells closely resembled Ad12-induced brain tumors. The transformed cell focus tended to appear near the embryonic brain cell (EB cell) or glioblastic cell (GB cell). The transformed cells were morphologically similar to the EB or GB cell. Some subcultured transformed cells showed a rosette-like pattern, and the surrounding space arrangement was similar to that of the ventricular wall. The incidence of brain cell transformations decreased with increased hamster age. This decreased incidence with age corresponded to the decreased numbers of EB or GB cells present in progressively older hamsters. From these results, it is concluded that the target cells of AD12 in hamster brain cell cultures are probably the EB or GB cells.


In vitro studies on target cells of oncogenic adenoviruses in hamster brain. III. In vitro transformation of brain cells of hamsters at various ages by bovine adenovirus type 3.

In vitro transformation of brain cells of hamsters at various ages was examined after the addition of bovine adenovirus type 3 to determine the type and origin of the target cells. Cellular transformations occurred only in cultures of fetus and newborn animals and at low incidences. Nine cell lines were obtained. Virus specific tumor antigens were demonstrated in the transformed cells. The present investigation suggested that bovine adenovirus type 3 might transform mesenchymal cells (ME cell) and that these cells are probably of meningeal or vascular origin. The histological picture of tumors following transplantation of the transformed cells resembled human primary sarcoma of the meninges and brain.


Changes in the levels of certain vitamins from B group under conditions of vitamin deficiency.

In experiments on rabbits the levels of vitamins B1, B2, B6, PP and pantothenic acid were determined to establish the grade and the sequence of their disappearance from the blood and liver in states of exogenous and endogenous vitamin deficiency. The experiments were carried out in which vitamin deficiency was produced by removing vitamins from the diet or by giving the rabbits antibiotics of different activity spectrum when the animals were kept on a diet without vitamins. Determinations of vitamin levels were performed using microbiological methods. It was found that the time after which changes in vitamin levels developed as well as the intensity of changes differ, depending on the experimental model used, but the sequence of vitamin disappearance from the organism is always the same. The first to disappear were vitamins B1, followed by vitamin B6 and pantothenic acid.


Inhibitory effects of glucocorticoids on prolactin release induced by thyrotropin-releasing hormone in man.

Glucocorticoid effect on thyrotropin-releasing hormone (TRH)-induced prolactin (PRL) release was studied in female patients with collagen or autoimmune diseases. Long-term, high dose glucocorticoid therapy tended to inhibit the response of plasma PRL to TRH. A negative correlation (r=-0.40) was found between the logarithm of total dose of glucocorticoids received and the magnitude of plasma PRL response to TRH (p less than .05).


Phosphorylation of purine and pyrimidine nucleosides by isolated rat liver mitochondria.

Formation of 5'-AMP, 5'-GMP, 5'-CMP and 5'UMP was confirmed in isolated rat liver mitochondria incubated with alpha-ketoglutarate, inorganic phosphate, purine nucleoside and pyrimidine nucleoside. Increased incorporation of 32Pi into ATP, GTP and UTP was observed by adding purine- and pyrimidine nucleosides. The phosphorylation of nucleosides was inhibited severely by arsenite and affected slightly by the addition of nuclear or post-mitochondrial fraction.


Cutaneous reactions to propranolol (Inderal).

Six patients wit psoriasiform cutaneous eruptions that developed during long-term therapy with propranolol (Inderal) have been studied. The cutaneous changes closely resembled those seen during treatment with practolol (Eraldin). The duration of treatment before the rash was recognized averaged 10 months, which is about the same latency period as in patients with practolol-induced exanthemas. The exanthemas disappeared gradually within 1-5 weeks after treatment with propranolol had been stopped. In 4 of 5 patients the skin eruptions reappeared within 2-4 days after oral challenge with propranolol. In the fifth patient, who developed a rash after challenge with practolol and severe abdominal colics after challenge with oxprenolol, no further provocation tests with propranolol were attempted. Skin biopsies obtained from 3 patients showed a microscopical picture similar to that seen in practolol exanthemas. The pathogenetic mechanism responsible for these adverse cutaneous reactions is unknown. However, since the possibility exists, that these changes may be caused by blockade of the epidermal beta-receptors, it is recommended that all patients receiving beta-blocking drugs should be examined carefully for similar adverse reactions. Special attention should be drawn to the reversible skin changes, since during treatment with practolol these have often preceded serious complications from other organs.


Chronic myocardial disease. I. Clinical picture related to long-term prognosis.

The prognosis in chronic myocardial disease is not well defined, partly because of a wide spectrum of clinical courses, and partly because of relatively short observation periods. This paper describes 106 patients followed for 2-12 years. Development or worsening of symptoms after intercurrent infections was associated with a more severe outlook than an insidious debut. The ability to develop myocardial hypertrophy appeared to be an important factor in deciding the prognosis. Pump failure was the cause of death in 80% of the patients, while 16% died suddenly. A favourable course was often noted in patients with ECG signs of left ventricular hypertrophy. This was also the case in patients who developed systemic hypertension. The presence of low voltage, especially in combination with left atrial enlargement, was associated with a malignant development.


Chronic myocardial disease. II. Haemodynamic findings related to long-term prognosis.

Haemodynamic findings in 106 patients with chronic myocardial disease have been related to the course of the disease during a follow-up period of 2-12 years. A high filling pressure, especially when combined with low cardiac output, suggested a severe development. The worst outlook was noted in patients who in addition had raised pulmonary arteriolar resistance. On the other hand, a low filling pressure even if cardiac output was reduced, was generally associated with a more benign course. An increase in left ventricular wall thickness as determined angiographically suggested a better outlook than normal wall thickness.


Vectorcardiographic, electrocardiographic, and angiographic correlations in apparently isolated inferior wall myocardial infarction.

Twenty-six patients with ECG evidence of localized inferior myocardial infarction and poor ejection fraction (less than 50 per cent) were compared with 26 patients with similar ECG's, but with normal ejection fraction (over 50 per cent). The poor ejection fraction group had significantly more frequent and more severe disease in left anterior descending artery and a higher incidence of triple coronary obstruction than the normal ejection fraction group. The poor ejection fraction group had a significantly greater incidence of ventricular asynergy in the anterior and apical segments of left ventricle. Vectorcardiography was available in 35 of the 52 patients studied and frequently supplied diagnostic information not available in the scalar ECG's. Of 18 patients with scalar ECG patterns of isols, vectorcardiography identified five cases with anterior infarction, three with left ventricular hypertrophy, and one with left anterior hemiblock. Vectorcardiography is a valuable supplementary tool in the clinical assessment of patients with apparently isolated inferior infarction. When extensive coronary and poor ventricular function exist, VCG clues may be expected in about half the patients.


Changes in heart dipole moment after surgical correction of atrial septal defect.

Spatial dipole moment of the heart (M) was measured in patients before and 1 to 2 weeks after repair of atrial septal defect. In normals three peaks of M were found, corresponding to excitation of the septum (M1), ventricular walls (M2), and basal portion of ventricles (M3). Most of the patients also had an M2a peak at 52 to 56 per cent of QRS duration. The patients with secundum defects were divided into two groups, A and B, depending on whether right ventricular pressure was less than or greater than 30 mm. Hg. M1 was smaller for the Group B patients than for the normals for Group A and decreased slightly after surgery. M2 was about half normal for Group A and one third normal for Group B. After surgery M2 increased for both Groups but was still below normal. There were small mean clockwise rotations of the M2 vector in the horizontal plane PO. The M2a vector pointed to the right anterior or posterior and generally downward. In three Group A patients the M2a peak disappeared PO but in two others it increased in magnitude with a counterclockwise rotation in the horizontal plane. In the Group B patients, M2a increased in all cases. The M3 vectors generally increased slightly in magnitude postoperatively. M3 pointed posteriorly in the normals but to the right in the patients, with little change in direction after surgery. Both P and T curves dropped in magnitude after surgery. The changes observed may be due in part to restoration of right ventricular blood volume towards normal after surgery. The remaining VCG abnormalities should be due to right ventricular hypertrophy.


Cardiomyopathy associated with amphetamine administration.

A 45-year-old woman with congestive heart failure, in whom there was no evidence of coronary heart disease, valve disease, or other demonstrable cause of heart failure, was found to have taken high doses of dextroamphetamine over a long period. Withdrawal of amphetamine resulted in deterioration, suggesting a physical cardiac dependence on the drug. The clinical and autopsy findings are presented and the similarities to the myocarditis associated with pheochromocytoma are discussed. The evidence presented suggests a causal relationship between administration of dextroamphetamine and the cardiomyopathy.


Echocardiographic evidence of outflow tract obstruction in Pompe's disease (glycogen storage disease of the heart).

A 7 month old black female infant with the clinical findings of Pompe's disease is presented. The diagnosis of an infiltrative myocardiopathy with left ventricular outflow tract obstruction presenting with a pronounced systolic anterior motion of the anterior mitral valve leaflet was made by echocardiography. This diagnosis was confirmed by cardiac catheterization and angiocardiography. Pathologic findings were consistent with Pompe's disease (type II glycogen storage disease). The presence of systolic anterior motion of the mitral valve in this patient suggests that this finding is not pathognomonic of idiopathic hypertrophic subaortic stenosis.


Mental retardation in a national population of young men in the Netherlands. I. Prevalence of severe mental retardation.

The prevalence of severe mental retardation, derived from military records, is analyzed in terms of time trend, region, religious affiliation and urbanization of birth place. The data are singular in that they are national and virtually complete for a total population of young men, 19-year-old survivors of male births in the Netherlands over the years 1944 through 1947, and they include variables not previously studied. Previous local studies are shown to agree well with the national prevalence of 3.7 per 1,000. Prevalence rose through the four birth years for all forms of severe mental retardation, but most markedly for Down's syndrome. In this condition agency surveys usually yeild equivalent rates to population surveys, and are an economical means of monitoring prevalence. Differences in rates are quite likely to indicate substantive differences. The remarkable similarities in rates from past surveys may reflect a high proportion of chromosomal and genetic abnormalities. The divergence between urban and rural rates over time demonstrated in this paper for four annual cohorts is attributed by inference to disparate increases in survival in different ecologic settings.


Laparoscopy in the obese patient.

A series of 49 patients weighing over 200 pounds is described, and a technique essential to successful care of obese patients is given. The ability to determine the intra-abdominal location of the Verres needle in order to create a physoperitoneum is the most important consideration.


Depressed mixed lymphocyte culture reactivity in mothers with recurrent spontaneous abortion.

One-way mixed wife-husband lymphocyte culture (MLC) reaction as well as human lymphocyte antigen (HLA) typing were performed on 29 couples, most of whom were infertile because of recurrent spontaneous abortion, and compared with 35 fertile couples with no abortion history. The 29 couples had been selected from 280 couples with a karyotyped spontaneous abortion. A significantly depressed MLD response (P=0.04) was observed in mothers of karyotypically normal abortuses when stimulated by the fathers but not when they were stimulated by males from the fertile couples. No significant difference in the frequency of HLA-A-and HLA-B-incompatible matings, respectively, was found between the abortion and the fertile couples. It is concluded that depressed maternal cellular response to paternal stimulation can be explained as a consequence of recurrent spontaneous abortion and that it is uncertain whether this depressed response plays a role in the etiology of recurrent abortion.


Relation between Na-K-ATPase activity and respiratory rate in the rat kidney.

The relation between Na-k-ATPase activity in homogenates of rat kidney and oxygen consumption in kidney slices was studied by employing different physiological maneuvers known to change the activity of renal Na-K-ATPase. Treatment of euthyroid rats with 3,5,3'-triiodo-1-thyronine increased Na-K-ATPase activity, sodium-dependent oxygen consumption (QO2[t]), and para-aminohippurate (PAH) accumulation by kidney slices without changing glomerular filtration rate or net sodium reabsorption by the intact kidney. Treatment with methylprednisolone also increased Na-K-ATPase, QO2[t], and PAH transport. Chronic potassium loading, on the other hand, increased renal Na-K-ATPase to the same degree as the first two procedures, but QO2[t] and PAH accumulation were unchanged. Partial nephrectomy induced an increase in the activity of Na-K-ATPase in homogenates of the remaining kidney fragment, but QO2[t] did not change significantly and PAH uptake was unaltered. An increase in the activity of Na-K-ATPase in kidney homogenates is therefore not necessarily associated with a parallel change in oxygen consumption by the intact cell.


Frey's syndrome.

Frey's syndrome occurs after parotid gland surgery or injury to the parotid gland and is characterized by gustatory sweating and erythema of the face upon mastication. The syndrome occurs in 50 to 60 per cent of all patients undergoing parotid surgery but the symptoms are only distressing to about 10 per cent of patients undergoing parotidectomy. A case of Frey's syndrome is reported  The patient obtained good relief of symptoms with scopolamine cream and atropine cream. Other methods of therapy are discussed along with their limitations. Noninvasive therapy with topical anticholinergic creams is effective and seems appropriate in the control of gustatory sweating.


Unrecognized myocardial infarction: five-year incidence, mortality, and risk factors.

In the course of 5 years, 9509 healthy adult subjects had an average annual incidence of 3.6 unrecognized infarcts per 1000 persons and 5.3 clinical ones per 1000 persons. A multivariate analysis showed that the most significant risk factors were age, left axis deviation, left ventricular hypertrophy, cigarette smoking, systolic or diastolic blood pressure, and peripheral vascular disease. Some of the known risk factors of clinical infarct, or angina pectoris or both, such as cholesterol, diabetes, anxiety, and psychosocial problems, do not play a significant role in unrecognized infarcts. Subjects whose electrocardiograms were initially interpreted by cardiologists as noninfarcts but by the computer as infarcts developed a high rate of unrecognized infarcts in the subsequent 5 years. A 7-year mortality follow-up showed a markedly higher rate among the unrecognized infarct group as compared with the noninfarct population, but significantly lower than those who developed a clinical infarct.


[Vohwinkel's syndrome (study of 4 cases)].

Four cases of keratoma hereditaria mutilans (Vohwinkel syndrome) are reported in two families, whereas clinic, metabolic, radiologic, histologic and cytogenetic aspects, the association with other processes, and therapeutic possibilities.


[Partial trisomy 21 (21q21 - 21q22.2)].

An abnormal chromosome 21 is reported in a child with a phenotype strongly reminiscent of trisomy 21 syndrome. It is shown to result from duplication of the segment 21q21 leads to 21q22.2. Comparison of the phenotype with that of other partial and total trisomics shows that the characteristic features of the trisomy 21 syndrome (mongolism), the mental retardation in particular - is due to trisomy 21q22.2 and perhaps 21q22.2.


DNA repair and frequency of x-ray and u.v.-light induced chromosome aberrations in leukocytes from patients with Down's syndrome.

DNA-repair and the frequency of chromosome aberration after u.v. and X-ray irradiation was studied on leukocytes from patients with Down's syndrome. The u.v.-induced DNA-repair synthesis was followed by the incorporation of [3H]thymidine in the presence of hydroxyurea. Similar dose-response curves were established for Down's syndrome leukocytes and controls. The cells from patients with Down's syndrome incorporated 70-75% of the activity of control cells at the various doses (32-196 erg/mm.2). This difference was significant for the two highest u.v.-doses (P less than 0-01). The yield of dicentric chromosomes after X-ray exposure (150 rad.) was 35% higher in Down's syndrome leukocytes than in the control cells (P less than 0-001). Combined u.v. and X-ray irradiation caused a twofold increase in the frequency of dicentric chromosomes in control cells, while the increase was only 27% in Down's syndrome leukocytes. This synergistic effect of u.v. and X-ray irradiation on the yield of dicentric chromosomes suggests that healing of X-ray and u.v.-induced DNA lesions may partly utilize the same repair enzymes. The results also indicate that DNA repair mechanisms are impaired in leukocytes from patients with Down's syndrome, which may contribute to the increased incidence of leukemia and the susceptibility to X-ray irradiation in this disorder.


Ages of death of children with Huntington's chorea and of their affected parents.

Bird et al. (1974) noted an interesting 'anticipation' phenomenon in Huntington's Chorea occurring in patients who inherited the gene from their father. More extensive samples from 165 pedigrees in the Low Countries permitted us to show that most of this apparent 'anticipation' is an artifact due to sampling biases related to the year of birth categories of the affected parents. When these biases were excluded no differences exist between the ages of death of affected mothers and their affected children, but a small difference of approximately two years remains between affected fathers and their affected children. This is explainable by the observation that juvenile cases of Huntington's Chorea usually have inherited the abnormality from their father.


The social effect of Huntington's chorea on reproductive effectiveness.

An attempt has been made to account for the observed departures from a model of a simple single-locus inheritance in Huntington's Chorea. It is postulated that an indirect effect of the abnormal allele lies in its social effects on individuals in the family group whether they bear the allele or not. This acts by influencing their mate-finding and reproductive behaviour so as to produce the observed statistical anomalies.


[Comparative study of the antibiotic sensitivity of Proteus hauseri bacteria belonging to different serological groups].

Sensitivity to 9 antibiotics of 1040 strains of Proteus belonging to the serological groups 03, 05, 06, 07, 010, 011, 013, 023, 024, 026, 027, 028 and 030 was studied. It was found that the above strains were sensitive and highly sensitive to the aminoglycosides and streptomycin, slightly sensitive to levomycetin and resistant to tetracyclines, erythromycin and penicillin. All the strains were polyresistant and 99.6 per cent of them were resistant to 4--9 antibiotics. Ten types of resistance were found. Proteus strains with the resistance type PETOtCht were most common. No relation between the occurrence of the strains of various serological groups and the character and level of their resistance to the antibiotics was found.


[Comparison of the sensitivity of pathogenic staphylococci isolated in 1974 to certain antibiotics and nitrofuran derivatives].

Sensitivity of 267 strains of pathogenic staphylococci isolated in 1974 was studied with respect to some antibiotics and nitrofuran derivatives by the method of serial dilutions on solid media. Sensitivity to penicillin, oxacillin, olemorphocycline, ristomycin and nitrofuran derivatives (furagin and salafur) was observed in 30.7 +/- 2.8, 61.8 +/-3, 29.2 +/-2.8 and 98.9 +/- 0.8 per cent of the cultures respectively.


[Antibiotic sensitivity of the Salmonella isolated from children in Krasnodar].

The results of the comparative study of antibiotic sensitivity of 138 Salmonella strains isolated from children patients in Krasnodar are presented. The isolates belonged to 9 serological types and were classified into 4 serogroups. Salmonella fo group B prevailed. The highest sensitivity was found with respect to neomycin. Strains sensitive to levomycetin and streptomycin were rare. Single strains were sensitive to all antibiotics tested. A tendency to polyresistance in ever increasing numbers of the strains was noted, which should be taken into consideration in treatment of salmonellesis in children with antibacterial drugs.


[Parameters of the toxic action of antibiotic derivatives of aureolic acid in acute and subchronic experiments].

The parameters of the lethal effect of aureolic acid derivatives, such as mithramycin, variamycin and olivomycin were studied on mice, rats and rabbits. As for the most of the administration routes the first two drugs were characterized by irregular distribution of resistance to thier lethal effect, which was mathematically expressed by polymodality of the dose-response curve. The above drugs were characterized by cumulative properties. The toxicity parameters depended on the animal species and administration route.


Heterogeneity of mouse B lymphocytes: studies using in vitro response to lipopolysaccharide.

When murine lymphoid cells from spleen and lymph nodes were cultured with LPS, all the transformed blasts observed were B cells bearing surface IgM which could be classified into two types depending upon whether they contained intracellular IgM (i mu +) or not (i mu -). At the ultrastructural level, the two categories of blasts appeared to correspond to cells having a well-developed ER or primarily free ribosomes. A striking observation was that the proportion of i mu + blasts among transformed cells was 5-10 times lower in cultures of lymph node than in cultures of spleen cells. The same observations were made on LPS cultures of "purified" B lymphocytes. Prolongation of the culture period did not modify these results. This indicated that the differences in the response to LPS reflected the existence of at least two pre-existing subpopulations of B cells unequally distributed in peripheral lymphoid organs. Analysis of the surface antigens and receptors (MBLA, surface Ig classes, Fc and C receptors) of the B cells from spleen and lymph nodes did show some differences between these two lymphoid organs. Among the transformed cells, Fc receptors were present mainly on i mu - cells and C receptors on i mu + cells; however, no strict correlation could be made between the i mu + or i mu - nature of these cells and their surface characteristics. The possibility that precursors of i mu + and i mu - blasts might represent virgin and memory B cells or B cell populations differing in their requirement for helper factors released by stimulated T cells was explored by using cultures of spleen and lymph node cells (1) obtained from germ-free mice, which are expected to have few memory cells, or (2) performed in the presence of the supernatant of mixed lymphocyte cultures, in conditions where these supernatants are known to contain factors released by activated T cells. Neither condition modified the degree of transformation or Ig content of blasts.


Porphyria-like cutaneous changes induced by tetracycline hydrochloride photosensitization.

Five patients manifested cutaneous changes indistinguishable from those noted in some porphyric disorders, consisting of fragility, denudation, and blister formation of sun-exposed skin. Microscopical examination showed subepidermal bulla formation and the desposition of PAS-positive, diastase-resistant material and IgG in or around the upper dermal blood vessel walls. There was also electron microscopical evidence of vascular basal lamina reduplication and the deposition of a fine fibrillar material in and around these vessels. However, no abnormal porphyrin formation was noted. All five patients had been receiving 250 mg of tetracycline hydrochloride twice a day for at least six months and had had extensive sun exposure prior to the onset of the condition. For four patients, discontinuing the medication led to complete remission, despite subsequent sun exposure; the fifth patient was much improved, but her skin was still somewhat fragile seven months later. We concluded that these cutaneous changes resulted from a low-grade photosensitization by tetracycline hydrochloride.


Topical vitamin A acid in the treatment of acne vulgaris (a controlled multicenter trial).

A controlled non-blind multicenter trial was conducted in 211 acne patients to test the activity of topical retinoic acid against sulfur-resorcinol--salicylic acid and placebo. Uniform evaluation criteria were used. After 8 weeks' treatment in comparable groups of patients, retinoic acid proved to be superior to the standard and to the placebo. The difference was statistically significant. Side effects were present in a number of patients treated with the active substances and with the placebo (mainly erythema), but rarely was the treatment discontinued.


Synthesis of glycosaminoglycan in adult human articular cartilage in organ culture from patients with rheumatoid arthritis.

Hyaline cartilage was obtained from patients undergoing synovectomy of the knee joint for rheumatoid arthritis. Eroded cartilage from beneath the invading pannus and relatively normal cartilage from the same joint were maintained in organ culture for three days. During the first 48 hours in culture the explants were exposed to 35SO4 in the medium. The equivalent layers of normal and eroded cartilage were analysed for DNA uronic acid and 35SO4 incorporation. There was a decrease in the DNA and uronic acid of the eroded cartilage, although only the latter reached statistical significance. The uptake of radioactive sulphate was significantly greater in explants taken from the eroded site than from normal areas. This increase in metabolic activity could well be a protective phenomenon.


Diminished mixed lymphocyte reaction in ankylosing spondylitis, relatives, and normal individuals all with HL-A 27.

The mixed lymphocyte reaction against a pool of blocked lymphocytes was studied in individuals with and without HL-A 27, including controls, spouses, parents, sibs, and offspring, and patients with ankylosing spondylitis. In controls without HL-A 27 the mean increment of counts perminute was 46,630 compared with 29,860 in asymptomatic controls and relatives with HL-A 27 and 26,277 in spondylitic patients with HL-A 27. The mean response was also reduced to 34,902 in sibs and offspring without HL-A 27, and to 20,916 in three patients with spondylitis in the absence of HL-A 27.


Experience with the DeBakey-surgitool aortic prosthetic valve.

The DeBakey-Surgitool aortic valve differs from popular cloth-covered prostheses by having a bare metal cage and a pyrolytic carbon ball. This valve was implanted in 37 patients. There were 7 operative deaths (19%), none of which was related to the valve design. Thirty survivors have been followed up to five years with 12 late deaths. The single thromboembolic event resulted in the only valve-related death. One patient developed a periprosthetic leak and another has hemolytic anemia. The incidence of thromboembolism in small series (3%) is comparable to that with the cloth-covered valves. This and the absence of complications related to cloth wear are important considerations when selecting a prosthesis.


Drugs and PGO waves in the lateral geniculate body of the curarized cat. I. PGO wave activity induced by Ro 43284 and by p-chlorophenylalanine (PCPA) as a basis for neuropharmacological studies.

Depletion, either predominantly of brain noradrenaline (NA) or preferentially of brain 5-hydroxytryptamine (5-HI), was achieved in cats by a single i.p. dose of 20 mg kg(-1) of Ro 4-1284, a benzoquinolizine derivative reducing the storage capacity of monoamine neurones, or by two i.p. injections of 300 mg kg(-1) of p-chlorophenylalanine (PCPA), an inhibitor of tryptophan hydroxylase. The ponto-geniculo-occipital (PGO) waves induced by these drugs (abbreviated PGO1284 and PGOPCPA, respectively) were continuously recorded from both lateral geniculate bodies of the unanaesthetized animals immobilized by gallamine. The present paper describes the characteristic qualitative and quantitative features of PGO1284 and PGOPCPA recorded under our standard experimental conditions; it provides the basis required for the following investigations of this series on the modulating effects of various drugs on this phasic brain activity. The density (number hr(-1) or number 0.5 hr(-1)) of PGO waves was inversely correlated with the levels of brain NA and 5-HT. At the peak of activity, during the second hr following the injection of Ro 4-1284, the density of PGO1284 was 630 hr(-1); it slightly declined during the following 5 hr. The highest activity of PGOPCPA (395 hr(-1)) appeared around the 74th hr after the first of 2 injections of PCPA; it remained constant for 3 hr and thereafter gradually declined. In constrst to the regular appearance of PGO1284, the PGOPCPA occurred in highly variable bursts. This difference was probably due to the different levels of NA and 5-HT in the brain.


[Activity and properties of adenosine triphosphatase in erythrocytes and various tissues (brain, liver, kidney, small intestine mucosa) of cattle].

In 15 samples of haemolysate of bovine erythrocytes, the splitting of phosphate from adenosine triphosphate average 158 +/- 63 X 10(-3) muMol/min/g haemolysate haemoglobin. Estimation of total adenosine triphosphatase in homogenates of various organs from cattle showed that spleen, liver, kidney and brain possessed high activity, while the activity was moderate in lung, myocardium and skeletal muscle, and low in endometrial mucosa and spinal cord. There was a relatively large proportion of Na-K-adenosine triphosphatase in brain and kidney. In various organs the activity of the enzyme was dependent upon the concentrations of Mg, Na, K and Ca. The inhibition of adenosine triphosphatase in various tissues by ouabain was studied. The optimum pH for the enzyme lay in the weakly alkaline region.


Laparoscopic sterilization in the immediate puerperium.

The safety and effectiveness of laparoscopic sterilization with electrocoagulation and separation of the tubes when performed within five days of a normal delivery is evaluated. The majority of patients (53.5%) were sterilized within 36 hours of delivery and were discharged on either the same day or the first post-sterilization day (91.0%). While some form of complications were reported for 9.5 percent of the patients, potentially serious complications occurred for only 2 patients (1.%). Most of the complications were probably unrelated to the laparoscopic procedure. At six months after sterilisation, complications were reported by 2.9 percent of the patients who returned for a follow-up visit. One patient became pregnant 3.5 months after sterilization. The results of this study suggest that performing laparoscopic sterilization in the immediate puerperium does not significantly increase the complication or failure rates of the procedure.


The excretion of 17-ketosteroids in patients suffering from mastopathia fibrocystica and mammary carcinomas.

Comparative investigations of the urinary excretion of different 17-ketosteroids in healthy women and patients suffering from mastopathia fibrocystica and/or mammary carcinomas had the following results: 1. A clear age dependency of the excretion of 17 ketosteroids during the regenerative period has been shown in all groups investigated, but it could not be observed during the postmenopausal phase. 2. There are no hints by the excretion values for a remarkable contribution of mammary carcinomas to the hormone metabolism of the host. 3. The excretion values of patients suffering from mastopathia fibrocystica confirm epidemiological results, which consider this from of mastopathia a precancerous one.


Health care utilization by persons with chronic disabilities who have been vocationally rehabilitated.

This investigation focuses on the effect of vocational rehabilitation on persons with chronic disabilities who have epilepsy, heart disease, spinal cord injury, arthritis, or emphysema. A thesis is proposed that such rehabilitants might be more vulnerable to unemployment than others, and it is also hypothesized that this frequently would be related to health care utilization. The study comprised 338 former clients of the State Rehabilitation Agency who had their cases successfully closed in the two-year period immediately prior to this study. All persons were sent a 12-page questionnaire dealing with matters of health, employment, social support, demography, medical economics, expressed needs, and relationship with the rehabilitation agency. Two-thirds (N = 213) of the clients responded. In addition, supplemental information was volunteered substantially by narrative and telephone. The primary thesis of the project was sustained; 43.7% were either unemployed or working less than full time six months after rehabilitation. However, few of the respondents reported having difficulties with health care utilization or availability. We propose a different rehabilitation services model to meet the needs of such clients. It should accentuate better diagnosis, careful selection of objectives, adequate vocational preparation, health care advisement, and more extensive job follow-up.


Reappraisal of skin flap closure for neonatal gastroschisis.

During the past 11 years, 18 infants with gastroschisis abdominal wall defects have undergone surgical repair at the UCLA Hospital. Sixteen infants had skin flap closure in infancy. A gastrostomy was performed on all infants, and peripheral intravenous hyperalimentation was used in 14 of the 18 infants. Sixteen of the 18 infants (89%) lived more than one year after surgical repair. Of these, 12 have undergone second-stage closure of the ventral hernia. Operative repair was greatly facilitated by forceful stretching of the abdominal musculature and milking of the bowel contents proximally into the stomach and distally out through the anus. The low morbidity and mortality of gastroschisis repair by primary skin closure, supplemented by intravensou hyperalimentation with late secondary ventral hernia repair, appear to justify continued use of this technique. Prosthetic materials probably should be reserved for reconstructing more complex abdominal wall defects.


Surgical treatment of infective valvular endocarditis.

Infective valvular endocarditis, whether occurring on a native or prosthetic heart valve, continues to carry a serious prognosis. Death is usually due to congestive heart failure, arrhythmia, or embolic complications, rather than infection per se. While antimicrobial therapy and medications to control congestive heart failure continue to be the treatment of choice in most cases, early valve replacement is indicated in certain situations. During the past four years, 12 patients underwent valve replacement for infective endocarditis. Six patients underwent elective valve replacement after antibiotic or antifugal therapy. All survived the operation and were improved. Six other patients underwent emergency valve replacement. Two patients died intraoperatively and their operations, in retrospect, were delayed unnecessarily. Four patients survived and are improved. Prosthetic valve replacement during the course of infective valvular endocarditis should be considered based on indications such as congestive heart failure, rhythm disturbances, etc, rather than the status of the infection.


[The structure of muscle spindles in the intercostal muscles and rectus abdominis of the human fetus].

The structure and innervation of muscle fibres were studied in external intercostal muscles and the streight muscle of the abdomen in 24--26-week human fetuses. The diameter of most spindles was shown to be within the range of 50--70 mkm, while in the streight muscle of the abdomen it could reach 100 mkm. In the external intercostal muscles the spindles, as a rule, were longer (300-500 mkm) than in the streight muscle of the abdomen (100--300 mkm) which was likely due to special functioning of the spindles in rhythmically working muscles. According to sensory innervation all the spindles may be divided into 3 main types: simple, intermediate and complex. In the external intercostal muscles there occur 3 types of spindles, while in the streight muscle of the abdomen there are spindles of a complex type.


[Histochemical characteristics of the lingual salivary glands of several classes of vertebrates].

The obtained results show that secretory elements of the tongue integuments submerged to the depth of the organ. It was accompanied by a formation and complication of terminal portions of the glands and their secretory pathways. In the process of evolution there occurred a divergence in the development of glandular cells from mucous ones in fishes through mucoserous and seromucous cells of amphibia and reptiles to mucous, seromucous and protein cells in the tongue glands of higher mammals. In mucous glands of terranian vertebrates, especially in mammals, the number of components in the composition of the produced secretion was found to increase.


[Treatment of diabetic retinopathy by centripetal pan-retinal photocoagulation with the argon laser].

The argon laser is such an effective method of treating diabetic retinopathy that it is used in the early stages. The indications for treatment are related either to ophthalmoscopically visible lesion, or to lesions revealed by angiography, which is necessary each year in all diabetics. As for as technique goes, it is desirable to begin pan-retinal photocoagulation in the periphery so as to take advantage of the drying out of the related oedema secondary to the destruction of the retina.


[Annular calcareous thesaurismosis of the cornea: Axenfeld's dystrophy].

The authors describe a case of primary corneal degeneration which is of calcareal nature and has annular shape. The patient has had a follow-up period of 15-16 years. A clinical study has been done and we have established a similitude with Axenfeld's primary calcareal degeneration, which was described by this author in 1917. Other corneal degenerations are also commented, which may induce into diagnostic error.


[Causes and consequences of orbital injuries].

For 20 years we have treated 219 patients from orbit trauma. According to the nature of injuries we had 57 cases with orbit fracture, 118 with the injury of intraorbital sof tissues, 37 with strange body in the orbit, and 7 cases in which orbital fistula had developed. Orbital fractures were most frequently caused by traffic accidents, until mutaual scuffles were the most frequent cause of injuries of the soft tissues of the orbit. As far as the localization of fractures is concerned, the worst disorder of sight and deformities in the orbital area were discovered in the lesion of the orbital roof and in multiple fractures of the orbital region.


[Electron microscopic study of trachoma].

The electron microscopic characteristics of the trachomatous conjunctiva and the trachoma agents and their evolution cycle are examined in a study on seven patients with Stage II trachoma, as classified by MacCallan, who had received no treatment. All the cellular epithelial structures remain intact despite the presence of pathological agents whose developmental cycle is entirely comparable to that observed "in vitro" in strain cultures. An imortant fact is the demonstration of macrophages in the sub-epithelial layers and of free pathological agents in the tarsal zone. The necrosis of the former may be responsible for the presence of the latter. A new hypothesis on the dynamics of trachoma infection is proposed together with its consequences in relationship to the forms of trachoma therapy.


[Dyshidrosis as a pacemaker for occupational dermatoses].

Out of 335 persons of less than 25 years, 236 had an intermittent dyshidrosis; half of them showed an allergic contact eczema of the hands. Microscopical and cultural investigations revealed a foot mycosis in 24 persons, and in 13 a candidiasis of the hands. Hence, dyshidrosis is obviously not of mycotic origin. 113 out of 150 sensitized persons of this group of age were dyshidrotics; 50% of them had not yet finished their apprenticeship. Obviously, dyshidrosis favours an early sensitization in professions in which allergens are frequently encountered.


Biochemical evidence for the reversed polarity of the outer membrane of the bacterial forespore.

Measurement of certain membrane-bound enzymic activities was used to study the orientation of the outer membrane of the double-membraned forespore of Bacillus megaterium KM. 2. Adenosine triphosphatase, NADH dehydrogenase and L-malate intact protoplasts, but were readily detected in intact stage II or IV forespores, consistent with reversed polarity of the outer forespore membrane relative to the mother-cell plasma membrane. 3. Measurement of NADH oxidase activity revealed that intact stage III forespores had the same high affinity for NADH as protoplast membrane preparations and protoplast lystates, consistent with ready access of NADH to oxidation sites on the outer forespores membrane. 4. Forespores and protoplasts showed osmometric behaviour in solutions of non-permanent solutes consistent with the presence of an intact permeability barrier in these structures.


Relation of C-6 glial cells in culture to myelin.

C-6 glial cells were shown to contain enriched in their plasma membranes an enzyme, 2':3'-cyclic nucleotide 3'-phosphohydrolase, characteristic of myelin, and, in addition, proteolipid protein and two basic proteins that are identical in their electrophoretic mobilities with the respective proteins found in myelin. The data indicate that C-6 cells exhibit features of myelin-producing glia as well as astrocytes.


Interaction between proteoglycan subunit and type II collagen from bovine nasal cartilage, and the preferential binding of proteoglycan subunit to type I collagen.

We studied the interaction of proteoglycan subunit with both types I and II collagen. All three molecular species were isolated from the ox. Type II collagen, prepared from papain-digested bovine nasal cartilage, was characterized by gel electrophoresis, amino acid analysis and CM-cellulose chromatography. By comparison of type I collagen, prepared from papain-digested calf skin, with native calf skin acid-soluble tropocollagen, we concluded that the papain treatment left the collagen molecules intact. Interactions were carried out at 4 degrees C in 0.06 M-sodium acetate, pH 4.8, and the results were studied by two slightly different methods involving CM-cellulose chromatography and polyacrylamide-gel electrophoresis. It was demonstrated that proteoglycan subunit, from bovine nasal cartilage, bound to cartilage collagen. Competitive-interaction experiments showed that, in the presence of equal amounts of calf skin acid-soluble tropocollagen (type I) and bovine nasal cartilage collagen (type II), proteoglycan subunit bound preferentially to the type I collagen. We suggest from these results that, although not measured under physiological conditions, it is unlikely that the binding in vivo between type II collagen and proteoglycan is appreciably stronger than that between type I collagen and proteoglycan.


The topographical location and unique nature of a glucokinase associated with the Golgi apparatus of rat liver.

A particulate glucokinase was recovered in the Golgi-rich fraction of rat liver prepared by the method of Morré [Methods Enzymol. (1971) 22, 130-148], thus extending the demonstration by Berthillier et al. [Biochim. Biophys. Acta (1973), 293, 370-378] of particulate glucokinase activity in a microsomal subfraction that showed enrichment in Golgi characteristics. The purity of this fraction was examined and it was then subjected to several treatments, the action of Triton X-100, freezing and thawing, and sonication to establish the topographical location of the glucokinase activity thus solubilized. The evidence suggests that the glucokinase activity is either soluble in the lumen of the Golgi apparatus or loosely associated with the inside of the Golgi membranes.


Persistent right atrial standstill.

An asymptomatic patient with cardiomegaly caused by isolated right atrial standstill is reported. The right atrium showed no evidence of contraction on pressure records or angiographically, while the left atrium functioned normally.


Placental metabolism of dehydroepiandrosterone sulphate in normal pregnancy.

An intravenous injection of 50 mg dehydroepiandrosterone was given to 19 women at the 38th week of pregnancy. The concentration of androstenedione, testosterone, oestradiol and oestrone in plasma was measured at intervals following the injection. The concentration of androstenedione and testosterone rose rapidly; reaching a peak after 10 minutes and returning to near baseline level by 30 minutes. Oestradiol rose more slowly, reaching a peak after 45 minutes and being sustained at high levels for 3 1/2 hours before returning slowly to baseline. Oestrone rose yet more slowly and did not reach a peak until 2 1/2 hours after the injection. It is concluded that the steroid responses are too variable from one individual to another for this test to be useful in the clinical assessment of fetal wellbeing but that it was likely to be a powerful tool in the investigation of placental steroid biogenesis.


The interaction of actin monomers with myosin heads and other muscle proteins.

The simplest interacting unit of actomyosin, viz., single myosin heads (subfragment 1) with actin monomers, has been studied at physiological ionic strength, by isolating the actin molecules from each other on a solid support. The interaction is characterized by a binding constant of 10(5) to 10(6) M-1 in the temperature range 4-30degrees C. It is endothermic with a standard enthalpy of 24 +/- 10 kcal mol-1, and a standard entropy of 110 +/- 40 eu. It is thus, like many protein-protein association processes, entropy-driven. Despite the high affinity of the association, which is comparable in its binding constant to that of subfragment 1 with F-actin, there is only very small activation of myosin ATPase. The ionic-strength dependence of the interaction shows unusual features. Binding of the proteins of the relaxing system to the monomeric actin was also examined: troponin binds both in the presence and absence of calcium ions, but neither tropomyosin nor the tropomyosin-troponin complex was found to bind significantly. Monomeric actin has also been examined as a function of ionic strength by spectroscopic methods; it appears that conformational differences between the G and the F state are the consequence of polymerization, and not of the change in ionic strength required to being the conversion about.


Retention of enzyme activity by detergent-solubilized sarcoplasmic Ca2+ -ATPase.

The Ca2+ -activated ATPase of sarcoplasmic reticulum can exist in true solution in the presence of some nonionic detergents, with retention of enzymatic activity for several days. The soluble active particles retain about 30 mol of phospholipid per mol of polypeptide chain even in the presence of a large excess of detergent, indicating the existence of relatively strong attractive forces between protein and lipid, as previous work from other laboratories has already suggested. Deoxycholate is much more effective than nonionic detergents in removing protein-bound lipid and, when used at solubilizing concentrations, completely delipidates and inactivates the ATPase. Preliminary molecular weight measurements indicate that the Ca2+ -ATPase exists as an oligomer in the native membrane: fully active enzyme in Tween 80 has a minimal protein molecular weight of about 400 000, corresponding to a trimer or tetramer of the ATPase polypeptide chain, and even the inactive enzyme in deoxycholate contains a substantial fraction of dimeric protein.


Isolation and properties of platelet myosin light chain kinase.

A protein kinase which phosphorylates the 20 000-dalton light chain of platelet myosin has been isolated from human blood platelets and purified approximately 600-fold. Elution of a 7.5% polyacrylamide gel following electrophoresis of the partially purified enzyme yielded a single peak of kinase activity which could be aligned with a protein band on  a stained gel. Assuming a globular shape, a native molecular weight of 83 000 (+/- 10%) was determined by gel filtration on Bio-Gel P-200. The kinase requires Mg2+ for activity and is not sensitive to the removal of trace Ca2+. The enzyme purified from human platelets phosphorylates the 20 000-dalton light chain of mouse fibroblast and chicken gizzard myosin, but does not phosphorylate human skeletal and cardiac myosin.


Inhibition of energy-transducing functions of chloroplast membranes by lipophilic iron chelators.

Lipophilic metal chelators inhibit various energy-transducing functions of chloroplasts. The following observations were made 1. Photophosphorylation coupled to any known mode of electron transfer, i.e. whole-chain noncyclic, the partial noncyclic Photosystem I or Photosystem II reactions, or cyclic, is inhibited by several lipophilic chelators, but not by hydrophilic chelators. 2. The light- and dithioerythritol-dependent Mg2+-ATPase was also inhibited by the lipophilic chelators. 3. Electron transport through either partial reaction. Photosystem I or Photosystem II was not inhibited by lipophilic chelators. Whole-chain coupled electron transport was inhibited by bathophenanthroline, and the inhibition was not reversed by uncouplers. The diketone chelators diphenyl propanedione and nonanedione inhibited the coupled, whole-chain electron transport and the inhibition was reversed by uncouplers, a pattern typical of energy transfer inhibitors. The electron transport inhibition site is localized in the region of platoquinone leads to cytochrome f. This inhibition site is consistent with other recent work (Prince et al. (1975) FEBS Lett. 51, 108 and Malkin and Aparicio (1975) Biochem. Biophys. Res. Commun. 63, 1157) showing that a non-heme iron protein is present in chloroplasts having a redox potential near + 290 mV. A likely position for such a component to function in electron transport would be between plastoquinone and cytochrome f. just where our data suggests there to be a functional metalloprotein. 4. Some of the lipophilic chelators induce H+ leakiness in the chloroplast membrane, making interpretation of their phosphorylation inhibition difficult. However, 1-3 mM nonanedione does not induce significant H+ leakiness, while inhibiting ATP formation and the Mg2+-ATPase. Nonanedione, at those concentrations, causes a two- to four-fold increase in the extent of H+ uptake. 5. These results are consistent with, but do not prove, the involvement of a non-heme iron or a metalloprotein in chloroplast energy transduction.


Ca2+ dependence of tension and ADP production in segments of chemically skinned muscle fibers.

Both ADP production and tension have been measured in segments of chemically skinned fibers contracting at different Ca2+ concentrations. Full mechanical activation occurred between pCa 7.00 and pCa 6.50. The total ATPase was due to both actomyosin and non-actomyosin ATPase. Actomyosin ATPase was observed at pCa 7.09 without accompanying tension. The Ca2+ dependence of tension was steeper than actomyosin ATPase. This finding implies some rate constants of the mechano-chemical cycle are Ca2+ dependent. Non-actomyosin ATPase was measured in fibers stretched beyond overlap of the thick and thin filaments. Sarcoplasmic reticulum was isolated and sarcoplasmic reticulum activity was measured in vitro under the same conditions as the single-fiber experiments. Non-actomyosin ATPase in the single fibers was found to be small compared to maximally activated actomyosin ATPase but larger than the ATPase that could be attributed to sarcoplasmic reticulum activity.


Effect of cytochalasin B on formation and properties of muscle F-actin.

Cytochalasin B stimulated polymerization and decreased the concentration of G-actin remaining in equilibrium with F-actin filaments. Polymerization in the presence of cytochalasin B gave rise to a smaller increase of viscosity but to the same increase in light scattering, compared to polymerization in the absence of cytochalasin B. Cytochalasin B reduced the viscosity of F-actin and caused the appearance of ATP hydrolysis by F-actin. The cytochalasin B-induced ATPase activity was inhibited by concentrations of KCl higher than 50 mM. The cytochalasin B-induced ATPase activity was enhanced by ethyleneglycol bis(alpha-aminoethyl ether)-N,N'-tetraacetic acid and reduced by MgCl2 at concentrations higher than 0.75 mM. The findings suggest that the stability of actin filaments is reduced by cytochalasin B.


Isolation of the basal and lateral plasma membranes of rat kidney tubule cells.

A method was developed to isolate renal basolateral membranes from cortical kidney tubule cells of single rats. The isolated membrane fraction was characterized by the measurement of marker enzyme activities and by electron microscopy. 1. After centrifugation of crude plasma membranes on a discontinuous sucrose density gradient the basolateral membranes accumulated at a sucrose density of p= 1.14-1.15 g/ml. The yield was 147 mug membrane protein/g kidney wet weight. Protein recovery was 0.1%. 2. (Na+ + K+)-ATPase was enriched 22-fold from the homogenate. The recovery was 2.6%. The (Na+ + K+)/Mg2+-ATPase ratio was 4.1. 3. The contamination by brush borders was small. Alkaline phosphatase was 1.6-fold enriched and 0.2% was recovered. Aminopeptidase was 1-fold enriched with a recovery of 0.1%. The contamination by mitochondria, lysosomes and endoplasmic reticulum was negligible. 4. In electron micrographs the basolateral membranes showed a typical triple layered profile and were characterized by the presence of junctional complexes, gap junctions or tight junctions.


Studies on phosphate transport in Escherichia coli. II. Effects of metabolic inhibitors and divalent cations.

1. Study has been made of the effects of a variety of metabolic inhibitors and divalent cations (Ni2+ and Mn2+), normally after 5 min exposure, on the biphasic uptake of inorganic phosphate (Pi) exhibited by phosphate-deprived cells of Escherichia coli, strains AB3311 (Reeves met-) and CBT302 (a (Ca2+ + Mg2+)-ATPase-deficient mutant). 2. In AB3311 cells cyanide (1-10 mM) produced comparable reductions in phosphate uptake to anaerobiosis, but in both instances significant uptake was maintained. Examination of intracellular Pi concentrations showed that, despite these inhibitions, Pi is still concentrated 130 times compared to 394 times under aerobic conditions. Arsenate (100 muM) and iodoacetate (100 muM pre-exposed 15 min) both abolished anaerobic-supported uptake. Under aerobic conditions the former eliminated primary uptake while the latter reduced both phases of uptake 60%. The uncouplers, dinitrophenol (100-1000 muM) and carbonyl cyanide m-chlorophenyl hydrazone (CCCP) (50muM) produced very significant, but not complete inhibitions of both phases of uptake. Inhibitions by iodoacetate and dinitrophenol were additive while dithiothreitol protected against the effects of 50-250 mum CCCP. N,N'-Dicyclo-hexylcarbodiimide (DCCD), the potent inhibitor of membrane-bound (Ca2+ + Mg2+)-ATPase, at 10(-3) M caused significant inhibitions of aerobic- (approx. 60%) and anaerobic- (approx. 80%) supported uptakes thus suggesting some obligatory requirement for this ATPase. 3. CBT302 cells, like AB3311, supported Pi transport both aerobically and anaerobically. CCCP (50muM) reduced the primary uptake similarly to AB3311 cells, but the secondary uptake was less affected. DCCD (10(-5)-10(-3) M), as expected, showed no effects in contrast to AB3311 cells. 4. In AB3311 cells Ni2+ (10 mM) caused significant but different reductions of secondary (70%) and primary (33%) phases of phosphate uptake. Mn2+ (10 mM) showed a greater differential effect with the primary uptake being minimally affected and the secondary uptake being abolished (97%). Partial relief of these inhibitions by Mg2+ (10 mM), suggested that these ions compete with Mg2+ transport. High voltage electrophoresis studies showed that Ni2+ cause intensification in the labelling from 32Pi (i.e. during Pi uptake) of hexose phosphates and a reduction in the labelling of complex molecules left at the origin. With Mn2+, labelling of fructose 1,6-diphosphate was reduced, the triose phosphate area was intensified and an unknown area (X) was intensely labelled. When Mn2+ was combined with anaerobiosis, phosphate uptake though diminished in rate exceeded after 16 min the plateau level of uptake under aerobic conditions with Mn2+ present.


Micrococcus lysodeikticus membrane ATPase. Effect of trypsin on stimulation of a purified form of the enzyme and idenfification of its natural inhibitor.

A soluble purified form of Micrococcus lysodeikticus ATPase (form BAT, from strain B, active, trypsin-stimulated) was stimulated 100% by trypsin and this stimulation was inhibited by preincubation of the protease with phenyl methyl sulphonylfluoride. This form of the enzyme was also stimulated 125-150% by filtration on Sephadex G-200. Analysis by sodium dodecyl sulphate-gel electrophoresis showed that stimulation of this form of M. lysodeikticus ATPase was always accompanied by the disappearance of a subunit of mol. wt. 25000 (epsilon subunit). It suggests that this subunit is the natural inhibitor of M. lysodeikticus ATPase. In the case of ATPase stimulation by trypsin, a partial and limited degradation of the alpha subunit was also observed. The interaction between the epsilon subunit and the rest of the ATPase complex was reversibly affected by pH, suggesting its non-covalent nature.


Isolation and partial characterization of chick brain synaptic plasma membranes.

An isolation procedure for synaptic plasma membranes from whole chick brain is reported that uses the combined flotation-sedimentation density gradient centrifugation procedure described by Jones and Matus (Jones, D. H. and Matus, A. I. (1974) Biochim. Biophys. Acta 356, 276-287) for rat brain. The particulate of the osmotically shocked and sonicated crude mitochondrial fraction was used for a flotation-sedimentation gradient step. Four fractions were recovered from the gradient after 30 min centrifugation. The fractions were identified and characterized by electron microscopy and by several markers for plasma membrane and other subcellular organelles. Fraction 2 was recovered from the 28.5-34% (w/v) sucrose interphase and contained the major part of the activities of the neuronal plasma membrane marker enzymes. The specific activities of the (Na+ +K+)-activated ATPase (EC 3.6.1.3), acetylcholinesterase (EC 3.1.1.7) and 5'-nucleotidase (EC 3.1.3.5) were, respectively, 4.5, 2.0 and 1.2 times higher than in the homogenate. However, Fraction 2 also contained considerable amounts of activities of putative lysosomal and microsomal markers in addition to lower amounts of mitochondrial and myelin markers. Although no prepurification of synaptosomes from the crude mitochondrial fraction was performed, the synaptic plasma membranes obtained showed many properties analogous to similar preparations from rat brain described in recent years.


Effects of ATP and magnesium ions on the fluorescence of harmala alkaloids. Restrictions for the use of harmala alkaloids as fluorescent probes for (Na+ + K+)-ATPase.

1. Harmine and harmaline were investigated as potentially useful fluorescent inhibitors of (Na+ + K+) activated ATPase. 29 From spectroscopic measurements both compounds were shown to form 1 : 1 complexes with ATP, the dissociation constants being 0.65 mM and 1.83 mM for harmine and harmaline respectively. Addition of Mg2+ and enzyme further affected these equilibria. 3. Although it was possible to demonstrate a competitive effect of harmine at the sodium-loading site of the enzyme, other inhibitory effects, including inhibitions of ouabain binding and the ouabain-insensitive ATPase were found. 4. It was concluded that the harmala alkaloids can inhibit (Na+ + K+)-activated ATPase in a complex way involving both Na- and ATP-binding sites. This severely limits their usefulness as spectroscopic probes.


[Inhibitors of the slow stage of proton transfer in the link connecting respiration with mitochondrial phosphorylation].

Electrophilic agents--derivatives of carbonic acids--are found to inhibit respiration, ATP synthesis and reverse electrone transport in intact mitochondria. The inhibition of respiration and ATPase was observed in intact mitochondria at 3 and 3u states (by Chance). Inhibitors concentrations, which caused 50% inhibition, were approximately the same. Sharp decrease of the effect of electrophilic inhibitors on respiration and ATPase activity in mitochondria and submitochondrial particles with substantially impaired coupling system was observed. The following conclusions are drawn on the basis of the data obtained: 1) electrophilic inhibitor attack the coupling site of respiration and ATP synthesis in mitochondria; 2) the reaction of the proton transport from the respiration proton pump to ATP synthetase is one of the slowest steps of the process of ATP-synthesis in mitochondria. A scheme of working the coupling system is suggested which includes the step of proton lateral diffusion.


[The effect of plasmin on kallikreinogen].

The effect of purified plasmin preparation on kallikreinogen isolated from rat blood plasma was studied by chromatography on DEAE-Sephadex. Plasmin was shown to affect kallikreinogen to form an active enzyme--kallikrein. The dependence of the activation degree on the plasmin activity and incubation time was studied. Evidence was found for the absence of Hagemann factor in the reaction medium, which excludes the possibility of indirect action of plasmin on kallikreinogen through the Hagemann factor fragments formed.


[Kinetic properties of skeletal muscle sarcolemmal Ca2+-ATPase].

Calcium activation of skeletal muscle sarcolemma Ca2+-ATPase is investigated. The investigation of a dependency of the initial rate of ATP hydrolysis on total concentration of substrate and on total and free calcium concentrations showed that the role of calcium ions is not limited by the formation of the substrate complex (CaATP2-). Calcium is absolutely necessary for the enzyme transition from inactive into active form. The inhibitory effect of free ATP is due to a decrease of free calcium concentration as a result of complexation with ATP, but not of competition with substrate in the active site. It is shown also that magnesium competitively inhibits the interaction of the enzyme with the substrate and non-competively suppress the activation of Ca2+-ATPase by free calcium.