[Fixation of various lectins at the surface of Crithidia luciliae].

Suspensions of Crithidia luciliae have been treated with 30 lectins: protozoans are agglutinated only by lectins inhibited with oses of structures I an II according to Mäkelä, and by lectins the site of fixation of which are unknown. The use of 5 lectins conjugated to fluorescein corroborate that lectins in congruity with group I and II, contrarily to those of group III, fasten upon the membrane and the flagella of Crithidia luciliae.


[Influence of intestinal microflora on the amino acid composition of lamb feces].

6 conventional and 5 germfree male lambs were fed ad libitum a UHT sterilized cow milk. Body weight and food intake were recorded. Whole feces were collected for 5 consecutive days. Growth rate reached 259 g/d for the germfree. Daily fecal excretion of dry matter and nitrogenous compounds are not found different in the two groups of animals. The influence of intestinal microflora appears on the biochemical composition of the feces. As compared to the conventional fecal proteins from germfree lambs are very high in threonine and serine and low in lysine. Moreover the difference of amino acid composition between these two groups come not only from the histidine alanine and arginine composition of bacteries; it also involves the high levels of threonine serine cystine and tyrosine of the endogenous digestive proteins.


[Appearance of phosphatase reticular cells in the spleen parenchyme of normal and tumor-bearing mice].

The splenic mouse parenchyma presents 3 reticular cells types revealed by histo-enzymatic technics: the phagocytic cells show a strong phosphatasic acid activity, the endothelial cells possess a phosphatasic alcalin or adenosine-triphosphatasic reaction, the perithelial cells are 5' nucleotidasic. These different cells are distributed by forming specific topographic structures in the splenic tissue. The phosphatasic alcalin reticular cells seem to be, with their distribution, characteristic elements of mouse spleen. Indeed the modifications in tumoral animal interest chiefly this cell category. In this case, the reticular cells form a deep and large membrane between marginal zone and perivascular lymphoid sheath of the white pulp. These different reticular cells probably react for the defense system of the animal.


[Relationship between the presence of microfilaments and the cell cycle of HeLa cells in synchronus culture].

Some cytoplasmic organelles have showed characteristic variations which are related to the different cell cycle phases, in thymidine synchonized HeLa cells in culture. In these cells, the most modified organelles were intracytoplasmic membranes (endoplasmic reticulum) and microfilament arrangements. Microfilaments were numerous under the cell membrane, but also some of them were dispersed in dense bundles. These structures were seen around the nucleus, 12-14 h after removal of excess thymidine (G1). They migrated to the periphery of the cell during S and G2. During mitosis, they were directly under superficial membrane-associated microfilaments.


[Evolvement of systems of serotonin uptake in rat hypothalamus during its development].

Kinetics of 3H serotonin accumulation into slices from hypothalamus have been compared in adult, puppy and foetus rat. In 15 days-old, as in adult rat, there are two components of 5 HT accumulation corresponding to the low and high affinity transport systems. For this latter, Km and Vmax values are much higher in adult than in 15 days old rat (in adult, Km=1,3 X 10(-7) and Vmax=0,33 X 10(-10); in 15 days old rat, Km=0,5 X 10(-7) and Vmax=0,125 X 10(-10)). On the opposite, in the 7 days old rat and in the 21 days old foetus, it is only possible to arbitrarely define one uptake system corresponding to the following apparent values: in the 7 days old rat, Km= 5 X 10(-7) and Vmax=2 X 10(-10), in the foetus, Km=0,2 X 10(-7) and Vmax=0,15 X 10(-10). These results showed important developmental differences in affinity of 3H serotonin to hypothalamus. The low and high affinity uptake systems existing in adult are only individualized in the 15 days old little rat.


[Antigenic and blood group activity of lamb gastric mucoproteins].

The antigenic properties of a lamb mucin and of a glycoprotein isolated from it are investigated. The proteins are studied by immunoelectrophoresis of the mucin and of the glycoprotein against either a sheep serum antiserum or an exclusively glycoprotein antiserum. No serum protein could be shown in the mucin. In addition, the glycoprotein gives a precipitin line only with the specific antiserum; its antigenic power seems to be concealed in the original mucin. The carbohydrates are studied through the investigation of the blood group activities by the hemagglutination inhibition technique. The mucin and the isolated glycoprotein show the same types of activities: they both prevent the agglutination of A1 and 0 red cells. The presence of two blood group activities in that glycoportein and its non-reactivity against A2 red cells corroborate the hypothesis that carbohydrate chains must be branched for the elicitation of these activities in a single molecule.


[Comparative study of the insulin-secreting effects of L (+) and of D (-) sodium lactate].

Both sodium 6(+) and D(-) lactate stimulate insulin secretion from the isolated perfused rat pancreas; however the insulin secretory effect of the L(+) form is much superior to that of the D(-) form. Glucagon secretion is not modified by the two isomers.


[Correlation between lactate output and insulin secretion of the isolated perfused rat pancreas under the influence of high concentrations of phenformin].

On the isolated perfused rat pancreas phenformin at high concentrations (10 mg/1, 50 mg/1 and 100 mg/1) provokes an increase of the insulin and lactate output in the effluent liquid. In no case is glucagon secretion modified by this substance. There exists a statistically significant correlations between the increase in insulin output and the increase in lactate output induced by phenformin.


[Adenosine triphosphate (ATP) and glucose. Action on insulin and glucagon secretion].

ATP stimulates glucagon and insulin secretions from the isolated perfused rat pancreas. This effect is modulated by glucose. Glucagon secretion is stimulated by ATP only in the absence or in the presence of a low glucose concentration (0.5 g/1). As to insulin secretion, it is strongly stimulated only in the presence of a glucose concentration of 1.5 g/1.


[Allometry and linear correlation coefficients binding the volumes of the different pituitary gland lobes, of the pituitary gland, of the subfornical organ and of the pineal gland to body weight and to hypothalamic volume in Rodents and in a Lagomorphe].

The morphometric study of the hypophysis, of the subfornical organ and of the pineal gland of 41 Rodents and one Lagomorphe shows that the anterior lobe of the hypophysis as well as the total hypophysis are better correlated to body weight than to the volume of the hypothalamus, while it is the contrary for the pars nervosa. The volume of the intermediate lobe varies very much from one species to another for the same body weight. The high value of the allometry coefficient of the pineal gland on somatic weight (1.25) is due to the fact that the heavy Rodents of our temperate climates have a large epiphysis while the small african Rodents possess a small pineal gland.


[Variations of the glucose 6 phosphodehydrogenase activity at the level of the diencephalon and of the gonads of the dormouse (Glis glis L.) during the annual cycle].

Activity levels of G6-PDH in the diencephalon and testes of Glis glis during the annual cycle show comparable variations. In the autumn, this activity is low and the gonads are inactive. At the anset of hibernation, this activity increases and becomes maximal during the period preceeding awakening. At mid-hibernation, the seminiferous tubules display in important enzymatic activity. Awakening is marked by a fall in enzymatic activity level that is a little more precosius of the diencephalon. The variations of enzymatic activity levels of G6-PDH in the diencephalon are similar in the female with an autumnal minimum and a maximal activity at the end of hibernation.


[Involution of gonadotrophic cells of the pituitary gland in the male frog, Rana esculenta, after section of the pituitary stem].

One and two months after section of the hypophyseal stalk in the male frog. Rana esculenta, and involution of numerous gonadotrophic cells, contrasting with the normal aspect of the eosinophilic cells can be observed.


[Action of 5-bromodeoxyuridine as a function of time on the aspect of chromosomes. Attempt at interpretation].

The different aspects of chromosomes in culture with B.U.D.R. are described. The times of B.U.D.R. action are 1/2, 1, 2 and 3 mitotic cycles. Four different aspects are obtained. For each time, 250 mitosis are studied, and classified. The results seem to confirm the semiconservative replication.


[Immunocytologic detection of peptides immunologically related to LRH and to SRIF in the garden dormouse under various conditions].

Immunocytologic technics have allowed us to detect appreciable and progressively increasing amounts of LRH and of SRIF in the median eminence of the dormouse during hibernation at 6 degrees C. In animals conditioned to darkness at 22 degrees C with the ovarian involution that is very important, there exists a nearly total disappearance of LRH while the amount of SRIF remains high. On the contrary permanent lighting at 22 degrees C responsible of an important reactivation of the genital sphere provoques a massive accumulation of LHR in the median eminence; SRIF is present in large amounts during the first weeks but diminishes of the experience is prolonged.


[Experimental study of the sensitizing power of methoxyflurane].

Methyoxyflurane, a halogenated general anaesthetic, showed a net sensibilizing effect in the guinea-pig when tested experimentally by the Magnusson and Klingman technique. According to these authors classification it may be classed as of "moderate allergenicity". In addition, the sensibilisation reaction obtained is prolonged. It remains at a similar level for 2 to 7 weeks after the end of the sensibilisation phase.


[Study of human pineal gland with scanning electron microscope and using Castaing's microsound].

Scanning electron microscopy of the human pineal gland showed two types of concretions, both which may be covered by micro-concretions, as well as the irregular surface of a cyst and group of formations considered as the endings of epiphyseal cell processes in a perivascular space.


[Correlations linking hypophysis and epiphysis volumes to body weight and brain weight in Simian and in man].

The human hypophyseal and epiphyseal volumes are quantitatively conformable to those of the simians studied. In simians and man, the hypophysis is better correlated to body weight and to brain weight than the epiphysis, as already observed by us in other Mammals (Insectivora, Prosimiano, Rodents and Chiroptera). The pineal gland must be correlated to a non essential somatic function that varies from one species to another at constant somatic weight.


[Reactional behavior of young crayfish towards metallic fons (Zn, Cu, Pb, and Cr): effect of temperature and sediments].

Reactional behaviour of two species of young crayfishes to different concentrations in chlorures (Zn, Cu, Pb and Cr) in option with breeding water is analysed. Discrimination capacity is more feeble with O. limosus. Physiological differences are confirmed by the raising of temperature or the presence of sediments. One interpretation of the ecological repartition of the two species A. pallipes and O. limosus is proposed on this basis.


[Compared effects of heavy metal ions (Cr+IV and Pb2+) on the respiratory intensity of 2 Crustacea Decapoda Brachyoura: Macropodia rostrata and Pachygrapsus marmoratus].

The effects on the crab's respiration of the sublethal metallic concentrations are analysed upon in toto organic structures and upon their very sensitive organs in the best and stressing conditions of temperature and of salinity. There is a connection between the metallic concentrations causing respiration deteriorations and the bearable metallic concentrations (starting lethal thresholds). In the same way there is a good correlation between the level of bioaccumulation and the metabolic depletion of the tissues. It's well proved by this research that a lethal metallic concentration in the best conditions of temperature and of salinity can greatly reduce the ability of the population to survive in the natural conditions of thermic and salin stress.


[Conditioning of slow electrocortical activity evoked by sensory stimulation and by a reflex movement in man].

Slow cerebral waves are recorded from 10 adults during an experiment consisting of the application of isolated or coupled sensorial stimulations, the weak sound occuring 880 msec prior to the strong light when coupled. Prior to coupling, the stimulations evoke on the vertex generally negative slow waves which would indicate an orientation reaction. After coupling, the responses to sound become constantly negative and are considered as waves described as "negative contingent variations". To the contrary, responses to light are inverted and become constantly positive. Such a phenomenon equally observed during experiments consisting of sound coupled to a reflex movement recalls the resolution of the negative contingent variation, the decision wave and the motor potentiel that accompanies the execution of voluntary movement, however here, it is produced during conditioning which does not require active motor participation by the subject. These results demonstrate that the simple coupling of two stimulations following the protocol developed by Pavlov provokes in man a complex collection of responses containing a motor component analogous to that which one observes in more elaborate experiments destined to prove the anticipation of the decision.


[Response of a membrane in a cochlear model to continuous and transient sounds].

Using enlarged mechanical models of the cochlea and true stimuli sinusoidal or transient, the authors give photographics documents which point out in objective way the "basilar" membrane responses. With pure tones, the membrane vibrates along its entire length no matter what the frequency. With transients, there is a maximum displacement in varying sites but always near the base.


[Electrophysiologic study of the cholinergic receptor of the myocardial membrane].

A double sucrose gap voltage clamp technic has been used to study the extra-current induced by acetylcholine (Iach) on the myocardial membrane on frog atrial trabeculae. I) No desensitization of the Iach current is noted for repeated perfusions of Ach. II) The Iach current is suppressed by atropine. III) The reversal potential Each is more negative than the resting potential --20 mV less than or equal to Each less than or equal to OmV.IV) The relationship Iach/Holding potential for various [K+]o shows a) That Each behaved as a potassium electrode, b) an inward going rectification. These results indicate that the cholinergic receptor might be related with the gk1 channel.


[Influence of estradiol on the capacity to produce antibodies during experimental amebiasis of the female golden hamster].

During experimental tissular amibiasis, the level of circulating antibodies, as detected by indirect immunofluorescence, is lower in female castrated golden hamsters who have been implanted with a pellet of oestradiol than in castrated control animals. Antibodies are first detected in the serum twenty four hours after infestation and their level becomes maximal near the fourteenth day.


[Observations on the mutagenic power of hycanthone in mammals].

Male mice from the C57Bl strain were given 150 mg/kg of hycanthone, an antischistosomial drug. Spermatocytes derived from the treated spermatogonia as well as the spermatocytes from the F1 male offspring sired the first three weeks after treatment were examined for the presence of chromosome rearrangements. Since no reciprocal translocation was recorded, it is concluded that treatment with hycanthone is not susceptible to produce transmissible chromosome anomalies in male mammalian germ cells.


[Effects of the cis-dichloro-diammino-platinum(II)-deoxyribonucleic acid on normal and cancerous cells].

The effects of cis-dichloro-diammino-platinum(II) (cis-Pt) bound to DNA have been compared to those of free cis-Pt in mouse Ehrlich tumour cells, in peritoneal macrophages and in chick embryo fibroblasts cultivated in vitro. Cis-Pt has no antimitotic activity anymore when linked to DNA. This would be due to the fact that free cis-Pt is not released from cis-Pt-DNA complex inside lysosomes.


[Annual protidic cycle and electrophoregrams of the savanna varan].

The circannual chronobiology of Varanus exanthematicus exanthematicus Bose, lacertilian reptile of Senegal, is characterized by a period of rest from January to June during the dry season and a period of activity from July to December during the wet season. The study of proteidemia and of the ratio of weight and height shows that the hydratation of corporal compartments is average from November to April and undergoes through changes from May to October. The electrophoresis of proteins shows the total disappearance of one of the fraction during the complete metabolic rest of the Varanus (from January to April). Finally the study of the electrophoresis of lipoproteins leads to the hypothesis that the Varanus can only make an insufficient biosynthesis of lipoproteins during the animal over feeding; besides this biosynthesis seems to be pratically stopped from January to May.


A second form of the single-strand specific endonuclease of Neurospora crassa which is associated with a double-strand exonuclease.

A second form of single-strand specific endonuclease, which is stable to heating up to 74 degrees C and does not bind strongly to phosphocellulose, has been partially purified from extracts of mycelia of wild-type Neurospora crassa. The endonuclease is associated with an equally heat-stable exonuclease which degrades linear but not circular double-stranded DNA and does not attack double-stranded RNA. The exonuclease probably also degrades single-stranded DNA. Both endonuclease and exonuclease activities are inhibited by 0.1-0.5 mM ATP. The exonuclease is preferentially inhibited by a variety of agents and preferentially inactivated by trypsin. A DNA-unwinding activity has also been detected in the nuclease preparation. Protease(s) present in the nuclease preparation destroy the DNA-unwinding and exonuclease activities on incubation at 37 degrees C, but do not affect the endonuclease activity. However, the heat-stability and chromatographic properties of the endonuclease are affected by this treatment. The altered properties of the endonuclease are very similar to those of the single-strand specific endonuclease which has been previously described. The combined nuclease activities of the unaltered preparational make up a putative recombination nuclease of N. crassa.


Pancreatic islet cell carcinoma with hypercalcemia and hypergastrinemia: response to streptozotocin.

A patient with metastatic islet cell carcinoma of the pancreas, recurrent peptic ulcer disease, and hypergastrinemia (Zollinger-Ellison syndrome) developed symptomatic hypercalcemia and renal insufficiency; she was treated with streptozotocin after parathyroidectomy failed to control her hypercalcemia. Shortly after somewhat less than the usual recommended dose of streptozotocin was administered, the serum calcium concentration fell to near normal with complete resolution of symptoms. Seven months after therapy, mild hypocalcemia, consistent with her degree of renal impairment was noted. However, mild hypercalcemia recurred 13 months after therapy. Shortly after streptozotocin therapy, the mean serum gastrin concentration fell to near normal with radiographic disappearance of the anastomotic ulcer. At 7 and 13 months after therapy, serum gastrin levels were normal. Streptozotocin therapy was accomplished without major complications; specifically, without a detrimental effect on the creatinine clearance. Thus, although hypercalcemia in patients with pancreatic islet cell tumors is often due to associated primary hyperparathyroidism, in some patients it may be due to secretion of a hypercalcemic substance from the tumor and may respond to streptozotocin. Similarly, hypergastrinemia in patients with islet cell tumors may also respond to streptozotocin.


Epithelioid sarcoma: case report with ultrastructural review, histogenetic discussion, and chemotherapeutic data.

A case of abdominal wall epithelioid sarcoma, studied by light and electron microscopy over a 3-year period, is presented. Ultrastructurally, there appear to be two types of tumor cells, light and dark, which differ by virtue of a heavier concentration of microfibrils and dilated rough endoplasmic reticulum in the dark cells. Both tumor cell types contain prominent Golgi systems, abundant free ribosomes, and numerous pinocytotic vesicles. The ultrastructural characteristics of the tumor cells resembel those of epithelioid cells of experimental human granulomas, as well as those of normal human synovium. A multifaceted relationship between histiocytes and synovial cells is demonstrated and it is concluded that the tumor is probably derived from mesenchymal reserve cells capable of differentiating a long histiocytic or synovial lines. Preliminary chemotherapeutic data are reviewed.


Surface membrane nucleoside triphosphatase activity and tumorigenicity of cultured liver epithelial cells.

A cell surface-located nucleoside triphosphatase activity can be assayed in liver epithelial cultures in situ with the incubation of intact cells in a medium containing [gamma-32P]adenosine triphosphate and correlated with the tumorigenicity of these cells in neonatal Wistar rats. The ectoenzyme activity of normal diploid cell lines is minimal, whereas a considerably high activity has been found in all tumorigenic cell lines tested. The optimum condition for the adenosinetriphosphatase activity is physiological with regard to osmolarity, ionic composition, pH, and substrate concentration in the medium. The enzyme is significantly stimulated by Ca2+, and its activation is controlled by Mg2+. Histochemical examinations indicate that glutaraldehyde-fixed cells of tumorigenic lines have Ca2+-stimulated adenosinetriphosphatase activity on the external surface. The isotopic assay of adenosine triphosphate hydrolysis by intact cells may provide a rapid method for screening oncogenesis in vitro of liver epithelial cells.


Purification of carcinoembryonic antigen by removal of contaminating mucopolysaccharides.

The fraction of carcinoembryonic antigen preparations not bound by concanavalin A was studied. A significant portion of this nonbound fraction of low antigenic activity was shown to be mucopolysaccharides by gas chromatography-mass spectrometry, cellulose acetate strip electrophoresis, and depolymerization by bovine testicular hyaluronidase.


The effect of cyclophosphamide and other drugs on the incidence of pulmonary metastases in mice.

A study was made of the effect of various cytotoxic drugs on the ability of i.v.-injected KHT sarcoma cells to form lung colonies in syngeneic C3H mice. Some enhancement of the number of lung colonies following an i.v. injection was seen following pretreatment of the mice with actinomycin D and mithramycin, while pretreatment with vinblastine, bleomycin, methotrexate, cytosine arabinoside, or 5-fluorouracil had little or no effect on lung colony formation. Pretreatment of the mice with cyclophosphamide, however, greatly increased lung colony formation (by a factor of approximately 100). This enhancement in lung colony formation was maximal when the drug was given 24 hr prio to the injection of tumor cells, but was seen as early as 2 hr and persisted as long as 8 weeks prior to the tumor cell injection. The degree of enhancement of lung colony formation was related to the dose of cyclophosphamide and was present in weaning as well as adult mice. This enhancement was not significantly reversed by anticoagulation with either aspirin or warfarin. Immunosuppression by whole-body irradiation did not affect the number of lung colonies seen in cyclophosphamide-treated mice. The mechanism by which cyclophosphamide enhances metastatic tumor growth within the lung is not known. The major effect, however, does not appear to be mediated  either by specific immunological or clotting factors.


Dehydroepiandrosterone sulfotransferase as a possible shunt for the control of steroid metabolism in human mammary carcinoma.

Human primary mammary tumors were examined to determine what factors were of importance in deciding relative rates of sulfurylation of dehydroepiandrosterone and 17beta-estradiol, such rates having been shown to correlate with the patient's prognosis and response to adrenalectomy (T. L. Dao and P.R. Libby. Enzymic Synthesis of Steroid Sulfate by Mammary Cancer and Its Clinical Implications. Natl. Cancer Inst. Monographs, 34: 205-210, 1971). The sulfurylation of dehydroepiandrosterone and 17beta-estradiol was studied in 41 tumors in vitro using tumor cytosol, adenosine triphosphate, [35S]SO42-, Mg2+, and added steroid. Six tumors showed no sulfurylating ability, 9 sulfurylated dehydroepiandrosterone at a rate greater than that for 17beta-estradiol (ratio, greater than 1), and 26 sulfurylated dehydroepiandrosterone at a rate lower than that for 17beta-estradiol (ratio, less than 1). Evidence was obtained that low levels of dehydroepiandrosterone sulfotransferase were responsible for ratios of less than 1, in many instances. Adenosine 3'-phosphate 5'-phosphosulfate synthesis and steroid sulfotransferase activities were measured in 30 tumors. A significant correlation was found between synthesis of the former and levels of estrogen sulfotransferase, but this relationship did not hold for dehydroepiandrosterone sulfotransferase, again due to low levels of this enzyme in many tumors. It is suggested that dehydroepiandrosterone sulfate formation in the tumors is mainly controlled by the sulfotransferase, which acts as a shunt in regulating the level of free dehydroepiandrosterone, and related compounds, available for metabolism to steroids influencing the growth of mammary epithelial cells.


Legislation concerning chemical carcinogens in several industrialized countries.

A survey was carried out on legislation in 14 industrialized countries relating to the prevention of occupational cancers. Two types of legislation were considered in particular: that dealing specifically with chemical carcinogens in the working environment, and that relating to compensation for occupational cancers. The survey revealed that legislation prohibiting the manufacture of chemicals known to be carcinogenic in humans or known to represent a possible cancer hazard to humans exists only in a limited number of the 14 countries considered and does not cover the same chemicals in each country. Legislation concerning monetary compensation is more common in these countreis than is legislation providing for primary prevention. There are two fundamental deficiencies in even the more comprehensive legislation. First, some chemicals for which carcinogenicity in humans has been proved are still produced in large quantities and are not covered by legislation. Second, the criteria used to determine which chemicals may be hazardous to humans when only experimental evidence of carcinogenicity exists are overexclusive, while the allowed concentrations of some of the chemicals recognized as possibly hazardous to humans appear to be very high.


The repair of X-ray induced chromosomal damage in trisomy 2-and normal diploid lymphocytes.

The frequency of chromosomal aberrations produced by X-rays is greater in lymphocytes cultured from trisomy 21 patients (Down's syndrome) than from normal diploid donors. This increase, which can be detected by a micronucleus assay for chromosomal damage, was postulated by us to result from a defect in the rejoining system which repairs chromosomal breaks. The postulated defect would result in a longer rejoining time, therapy permitting more movement of broken ends and thus enhancing the frequency of exchanges. To test this possibility, the time required for the rejoining (repair) of chromosome breaks was measured in lymphocytes from five Down's syndrome (four trisomy 21 and one D/G translocation partial trisomy 21) donors, from a monosomy 21 donor, and from five diploid donors. The rejoining time was reduced in the Down's syndrome lymphocytes in comparison to the normal diploid and monosomy 21 lymphocytes. Thus the repair of chromosome breaks, far from being defective as evidenced by a longer rejoining time in Down's syndrome cells, occurred more rapidly than in normal cells. A mechanism is proposed by which reduced rejoining times would increase aberration frequencies as a consequence of competition between a (hypothetical) error-free repair system and the error-prone repair system that generates chromosomal aberrations. We suggest that the alteration in the rejoining of chromosomal aberrations may underlie the increased susceptibility of people with Down's syndrome to leukemia.


Thalicarpine (NSC-68075): plasma decay and urinary excretion in man.

The plasma decay and urinary excretion of 3H-thalicarpine were studied in a total of 19 patients at doses of 300-1900 mg/m2. Plasma decay was triexponential with a terminal phase half-life of 198-1386 hours in five patients. Urinary excretion was slow and erratic, a total of 19.84% of the dose being excreted in 285 hours in nine patients. Only small amounts of unchanged thalicarpine appeared in the cerebrospinal fluid. The data indicate extensive localization of thalicarpine in the tissues and the danger of drug accumulation if it is given at short dose intervals.


Ehrlich ascites tumor (EAT) chalone effects on nascent DNA synthesis and DNA polymerase alpha and beta.

EAT chalone effects on nascent DNA synthesis and DNA polymerase were examined. Concentration related inhibition of 3H-thymidine (3H-TdR) incorporation into EAT cell DNA was noted over a chalone range of 50-200 mug/ml. RNA synthesis was not affected, but protein synthesis decreased an average of 82% during 3 hr. Nascent DNA pulse-labeled for 2 min was normally incorporated into bulk DNA in the presence of chalone, but crude alpha- and beta-polymerase activities were inhibited. Crude DNA polymerase for C3H mouse kidney and spleen was also partially inhibited by EAT chalone, suggesting non-specific inhibition of DNA polymerase. Preincubation studies of chalone with crude EAT DNA polymerase or 'gapped' DNA primer had no effect on chalone activity. Chalone may control mitotic activity by inhibiting alpha- and beta-polymerase activity, thereby decreasing nascent DNA synthesis. Nascent DNA is incorporated normally into bulk DNA in the presence of chalone, indicating the DNA ligase is not inhibited.


The mechanical characteristics of hypertrophied rabbit cardiac muscle in the absence of congestive heart failure: the contractile and series elastic elements.

Right ventricular papillary muscles from normal rabbits and rabbits with sustained pulmonary artery constriction (67% decrease in external diameter) were studied at several resting muscle lengths and at an early instant in the isometric twitch. Instantaneous force-velocity data were obtained at 30-38% of time to peak tension (TPT) and at 96%, 98%, and 100% of the resting muscle length at which active twitch tension was maximal. Unloaded shortening velocity (Vmax) was estimated with a linearized form of the Hill hyperbolic formula, and was depressed in hypertrophy to 36% less than normal. We found that Vmax did not change with muscle length in the normal or hypertrophied muscles; therefore there was a length- and time-independent depression of contractile element shortening capacity that was consistent with previous work from this laboratory which demonstrated a depression of myosin and actomyosin ATPase activity in hypertrophy.


Recuperative potential of cardiac muscle following relief of pressure overload hypertrophy and right ventricular failure in the cat.

This study examined the recuperative potential of cat hearts subjected to experimental right ventricular pressure overload (for a 10- to 14-day period) which provoked hypertrophy with and without congestive heart failure. Five groups of cats were studied: normal controls; one group with 70% pulmonary artery constriction which produced right ventricular hypertrophy (RVH); one group with an 87% constriction which also produced right ventricular hypertrophy but with congestive heart failure (CHF); and two groups which had been similarly subjected to pressure overload but which had been allowed a recovery period of 30 days after relief of the pressure overload. Both the 70% and 87% pulmonic constrictions were associated with extensive right ventricular hypertrophy, depression of myocardial contractile function, and severe redlction of cardiac norepinephrine stores (normal, 1.42 mug/g: RVH, 0.11 mug/g; CHF, 0.01 mug/g). After a 30-day period of relief from the pulmonic constriction normal hemodynamic function returned. In cats in which RVH had been relieved, right ventricular weight and contractile function were normal but catecholamine depletion persisted. Cats with relieved CHF showed depressed contractile function and depleted myocardial norepinephrine, and the right ventricular weight did not return to normal. Cardiac muscle of all pressure-overloaded nonrelieved hearts showed depressed velocity of shortening and depressed ability to sustain load. Cats with RVH alone regained normal muscle shortening velocity and load-bearing ability after relief. However, cardiac muscle from the CHF-relieved group recovered only unloaded shortening velocity while the ability to sustain load remained depressed. We conclude that the recuperative potential of myocardium damaged by pressure overload is adequate provided congestive heart failure has not occurred. Heart failure produces a persistent reduction in force-generating ability of the myocardium. Hypertrophy due to pressure overload, with or without CHF, leads to cardiac catecholamine depletion which is not readily reversed by relief of the overload.


A possible molecular mechanism of the action of digitalis: ouabain action on calcium binding to sites associated with a purified sodium-potassium-activated adenosine triphosphatase from kidney.

Calcium binding at 0 degrees C to a purified sheep kidney Na+,K+-ATPase was described by linear Scatchard plots. Binding at saturating free calcium was 65-80 nmol/mg of protein, or 30-40 mol of calcium/mol of enzyme. Aqueous emulsions of lipids extracted from Na+,K+-ATPase yielded dissociation constants and maximum calcium-binding values that were similar to those for native Na+,K+-ATPase. Phospholipase A treatment markedly reduced calcium binding. Pretreatment of native Na+,K+-ATPase with ouabain increased the dissociation constant for calcium binding from 131 +/- 7 to 192 +/- 7 muM without altering maximum calcium binding. Ouabain pretreatment did not affect calcium binding to extracted phospholipids, ouabain-insensitive ATPases, or heat denatured Na+,K+-ATPase, Na+ and K+ (5-20 mM) increased the dissociation constants for calcium, which suggests competition between the monovalent cations and calcium for the binding sites. At higher concentrations of monovalent cations, ouabain increased the apparent affinity of binding sites for calcium. Extrapolation to physiological cation concentrations revealed that the ouabain-induced increase in apparent affinity for calcium may be as much as 2- to 3-fold. These results suggest: (1) calcium binds to phospholipids associated with Na+,K+-ATPase; (2) ouabain interaction with Na+,K+-ATPase induces a perturbation that is transmitted to adjacent phospholipids, altering their affinity for calcium; and (3) at physiological concentrations of Na+ or K+, or both, ouabain interaction with Na+,K+-ATPase may lead to an increased pool of membrane-bound calcium.


An assay for total hexosamine and a differential assay for glucosamine and galactosamine.

Two new procedures are presented for quantitative determination of glucosamine and galactosamine. One, which is proposed for total hexosamine, yields chromogens of equal intensity with equal concentration of glucosamine and galactosamine. There is addition of the correspondent chromogens when they are present in mixtures. The procedure is presented as a manual as well as an automated assay. The other procedure is a differential assay which allows the detection of galactosamine without interference by glucosamine. By the two procedures, the hexosamines present in acid mucopolysaccharides and/or glycoproteins can be determined.


Transport ATPases of cardiac sarcolemma in experimental uremia.

Uremia was induced in rats and rabbits by unilateral nephrectomy and partial resection of the contralateral kidney. The effectiveness of the procedure was checked by determining serum urea levels that 7 days after operation were found to be more than twice as high in the operated than in the sham operated animals. Serum electrolytes at this time did not show significant alterations. In the cardiac sarcolemma from both kinds of animals the specific (Na+,K+)-ATPase activity was significantly decreased in the uremic animals. The basic, Mg2+-dependent ATPase was not affected.


Macrophage clearance function and immune complex disease in New Zealand Black/White F1 hybrid mice.

Macrophage clearance function in NZB, NZW, NZB/W, Ajax and B10D2 new mice was assessed by measurement of the rate of clearance (KPVP) of intravenously-injected 125I-labelled polyvinylpyrrolidone (PVP). There were significant strain and age-related variations in KPVP. In particular there was a marked fall in KPVP in NZB/W mice with increasing age. This fall was most apparent in female NZB/W and preceded the age at which renal disease usually develops in these animals. We suggest that ineffective macrophage function and production of low affinity antibody contribute to the early development of immune complex glomerulonephritis in these mice.


Concanavalin A-activated suppressor cells in normal human peripheral blood lymphocytes.

A 24-hr preincubation of human peripheral blood lymphocytes in the presence of Con A renders them suppressive for the response of untreated cells to soluble antigens, allogeneic cells and Con A.


Interdependence of Na-excretion, plasma electrolytes, plasma volume and renal Na-K-ATPase-activity in hypertensive rats.

Hypertensive Goldblatt-rats have higher than normal Na-appetite and an enhanced Na-output. They have normal plasma Na- and K-concentration and kidney weight but a significantly reduced plasma volume. The amount of renal membrane protein and the renal Na-K-ATPase-activity of hypertensive rats was found to be significantly below that of controls. In order to evaluate the role of Na-appetite, Na-excretion rate and renal Na-K-ATPase-activity in the electrolyte balance, Goldblatt-rats with a stable hypertension and control animals were put for 8 days on a Na-free diet. Na-excretion rate of control rats reached a minimum (13 muEq/100 g x 24 hr) within 5 days and was maintained on this level up to the end of the experiment. Na-free diet did not alter either the kidney weight or the amount of membrane protein of the animals. However, in salt-free fed control rats total renal Na-K-ATPase-activity was found elevated by about 10% as compared to animals maintained on normal diet. Goldblatt-rats continuously excreted significantly higher amounts of Na (35 muEq/100 g x 24 hr), had sharply reduced plasma volume and plasma Na- concentration. The renal Na-K-ATPase-activity should no adaptation in gold blatt-rats. In all animals studied the rate of Na-excretion showed a close indirect correlation with the renal Na-K-ATPase-activity. It is concluded, that Goldblatt-rats depend on dietary Na to a higher extent than controls because of their reduced capacity to retain Na. The increased Na-appetite of hypertensive rats is a factor secondary to Na-loss.


The function of (Na+, K+)-ATPase in the thick ascending limb of Henles loop.

The content and the properties of (Na+, K+)-ATPase in the thick ascending limb of the loop of Henle (TAL) are related to the rate and the characteristics of the transport of NaCl which has been measured in the isolated perfused cortical portion of the TAL by Burg and Green (Am. J. Physiol, 224, 659, 1973) and in the medullary portion of the TAL by Rocha and Kokko (J. Clin. Invest. 52, 612, 1973). It is concluded that the ouabain-sensitive, active transport of NaCl across the epithelium consists of primary active transport of Na and secondary active transport of Cl and that it is driven by the sodium pump or (Na+, K+)-ATPase.


The effect of Ca ion antagonist verapamil on ouabain inhibition of renal sodium reabsorption. Studies in the isolated perfused rat kidney.

Evaluation of a possible role of NaK-ATPase in transtubular sodium reabsorption is difficult, since ouabain doses that inhibit this enzyme system in vitro completely [1, 11], cannot be applied in vivo. Thus studies in the isolated perfused rat kidney were carried out [3, 11, 13, 15]. However, in this model, supramaximal doses of ouabain as used to block NaK-ATPase completely induced a potent vasoconstriction, which lowers the filtered load of sodium. Thus, a meaningful quantitative comparison of sodium transport in control and in experimental phases of ouabain inhibition has not been possible. At submaximal doses, in which filtered Na-loads are still comparable, transport activity was only reduced to approximately 50% [15]. In the following experiments we reinvestigated the relationship between inhibition of renal NaK-ATPase and the reduction of Na-reabsorption by ouabain under more appropriate conditions. Previously we have observed that verapamil (syn.: iproveratril, Isoptin), a Ca ion antagonist [2, 4, 6], effectively prevents autoregulation, and a fact which is pertinent for the present experiments, blocks this vasoconstrictive action of ouabain. Thus, using the Ca antagonist verapamil, it was possible to evaluate the inhibiting effect of this glycoside on renal sodium transport quantitatively without the hazards introduced by ouabain vasoconstriction which by the same token lowers filtered load.


Echocardiographic patterns in scleroderma.

The echocardiograms of two patients with sclerodermatous cardiac disease are described. In one patient the pattern was that of a congestive cardiomyopathy with ventricular dilatation and reduced wall motion. In the second patient the pattern was that of an infiltrative cardiomyopathy with thickened walls and reduced wall motion in the absence of ventricular dilatation. Echocardiographic studies are useful in the early detection of pericardial involvement and primary or secondary myocardial involvement by scleroderma and in following the progression of the disease process.


The relationship between affective and cognitive development in Down's Syndrome infants.

A close association between affective expression and cognitive development was demonstrated in a longitudinal study of 14 Down's syndrome infants. It was found that the Down's syndrome infants laughed to groups of stimulus items in the same order as did previous samples of normal infants. Although the process was delayed by several months, the retarded babies too laughed first at physically intrusive items and only later to items calling for greater cognitive sophistication. In addition, cognitive developmental status, assessed by the Bayley and Uzgiris-Hunt scales, paralleled and was predicted by the level of affective development. Predictive and concurrent correlations between Bayley mental scores and various indices of affectivity ranged from .68 to .92. There was striking individual consistency across affective, mental, and motor measures, suggesting the organized nature of retarded development. Finally, since Down's syndrome infants frequently smiled under conditions when normal babies would laugh, a role for tension production, in addition to cognitive factors, was suggested in accounting for the behavior of these infants.


Reverse fluorescent chromosome banding with chromomycin and DAPI.

Two DNA binding guanine-specific antibiotics, chromomycin A3 (CMA) and the closely related mithramycin (MM), were used as chromosome fluorescent dyes. Root-tip metaphase chromosomes of three plant species and human metaphase chromosomes were sequentially stained with CMA or MM and the DNA binding AT-specific fluorochrome 4'-6-diamidino-2-phenylindole (DAPI). In some cases a non-fluorescent counterstain was used as contrasting agent: methyl green in conjunction with CMA, and actinomycin D (AMD) in combination with DAPI.--In all three plant species, Vicia faba, Scilla siberica, and Ornithogalum caudatum, the nucleolus organiser regions and/or associated heterochromatin displayed very bright fluorescence with CMA and MM and, in general, heterochromatic segments (C-bands) which were bright with CMA and MM were pale with DAPI whereas segments which were dim with CMA and MM displayed very bright fluorescence with DAPI.--Human metaphase chromosomes showed a small longitudinal differentiation in CMA fluorescence, which was essentially the reverse of the banding pattern obtained with AMD/DAPI double-staining, but of lower contrast. The cma-banding pattern appears to be similar to the pattern found by R-banding procedures.


Insulin-treatment of diabetic rats: effects on duodenal calcium absorption.

We tested the hypothesis that depressed duodenal calcium absorption in the streptozotocin diabetic rat is the consequence of diabetes rather than nephrotoxicity of the diabetogenic agent causing abnormal renal vitamin D metabolism. We treated streptozotocin diabetic rats with insulin and compared their duodenal calcium transport response with that of untreated diabetics and matched controls. Insulin treatment restored depressed calcium transport of diabetics to control levels in in vivo studies and significantly increased calcium transport in vitro. Previous studies showed that even in uncontrolled diabetes the mucosa retains the ability to respond to an end organ stimulus enhancing calcium transport: 1,25-dihydroxycholecalciferol corrects the defect, but vitamin D and 25-hydroxycholecalciferol are ineffective. Since 1,25-dihydroxycholecalciferol is synthesized in the kidney, these findings, in conjunction with the current study, are consistent with the association of experimental diabetes with a renal defect depressing synthesis of 1,25-dihydroxycholecalciferol. Since insulin treatment restores duodenal calcium transport, the renal defects is unlikely to be caused by streptozotocin nephrotoxicity.


Role of testosterone and its metabolites in the differentiation of the mammary gland in rats.

The capacity of various androgens to virilize the differentiating mammary gland in the female rat fetus has been determined. Testosterone, 5alpha-androstane-3alpha, 17beta-diol (3alpha-diol), and dihydrotestosterone (DHT) virilize the anlagen of the mammary gland by suppressing nipple formation but 5alpha-androstane-3beta, 17beta-diol, androsterone, and dehydroepiandrosterone sulfate do not affect female mammary differentiation. However, unlike the genitalia and wolffian ducts of the female rat fetus in which the masculinizing potency of DHT and 3alpha-diol is greater than that of testosterone, testosterone is more potent than its metabolites DHT and 3alpha-diol, in virilizing the mammary gland. The results suggest that testosterone is the fetal androgen mediating masculine development of the mammary gland.


Steroids in fluids and sperm entering and leaving the bovine epididymis, epididymal tissue, and accessory sex gland secretions.

Ten steroids which may have a role in the process of sperm maturation within the epididymis were quantified by competitive protein binding or radioimmunoassay. Rete testis fluid (RTF) carrying testicular sperm into the epididymis was rich  in dehydroepiandrosterone and testosterone (21 +/- 2 and 33 +/- 3 ng/ml) while cauda eipididymal plasma (CEP) around sperm which have completed maturation had high levels of progesterone, dihydrotestosterone, 3beta-androstanediol, dehydroepiandrosterone and testosterone (7.4 +/- 0.8, 20.3 +/- 1.1, 6.5 +/- 0.4, 8.0 +/- 0.7 and 11.5 +/- 0.7 ng/ml). About 4 mug of steroids enter the epidymis daily in RTF, but less than 1% was found in CEP; the balance presumably was absorbed by the epithelium in the proximal caput epididymidis. Nevertheless, tissue levels of total 17beta-OH androgens were lower in the proximal caput than in the distal caput or corpus epididymidis. In all zones of the epididymis, dihydrotestosterone accounted fro about 70% of the total 17beta-OH androgens found in the nuclear fraction. In the cytoplasmic fraction, however, dihydrotestosterone predominated only in the distal caput and corpus epididymidis. In the cauda epididymidis, CEP and sperm probably accounted for less than 35% of the total 17beta-OH androgens and less than 25% of the dihydrotesterone. The progesterone concentration of the cauda than in the caput epidymidis. Twice washed testicular sperm contained more testosterone than cauda epididymal or ejaculated sperm (16.6 +/- 1.9, 1.6 +/- 0.2 and 1.5 +/- 0.3 ng/10(9) sperm, respectively), but less progesterone (0.5 +/- 0.1, 1.3 +/- 0.2 and 1.0 +/- 0.4 ng/10(9) sperm, respectively). As a consequence of mixture with estrogen-rich prostatic fluid (150 +/- 9 pg/ml), ejaculated sperm contained a relatively high amount of estrogens (112 +/- 15 pg/10(9) sperm). These studies revealed marked differences in steroid profiles of fluids entering and leaving the epididymis and of infertile testicular and fertile cauda epididymal sperm.


Blockade of the tonic hindlimb extensor component of maximal electroshock and pentylenetetrazol-induced seizures by drugs acting on muscle and muscle spindle systems: a perspective on method.

Blockade of tonic hindlimb extension (THE) in the maximal electroshock seizure test was demonstrated for C-28'882-Ba, a muscle spindle suppressant, chlorpromazine, a depressant of fusimotor drive, and gallamine, a neuromuscular blocking drug. These agents also blocked THE produced by very large doses of pentylenetetrazol. Dantrolene, a muscle contraction antagonist, and MI-65-S, a muscle spindle suppressant, significantly delayed THE. The data indicate that blockade of THE may be effected at a multiplicity of loci and may not be an expression of an "anticonvulsant" effect.


Use of the tissue slice technique for evaluation of renal transport processes.

A detailed discussion of the tissue slice technique for evaluation of transport phenomena is presented. Information is given concerning the preparation of tissue slices and the advantages of this procedure over corresponding in vivo techniques. In addition, the relationship of the in vitro renal transport of organic substances to in vivo renal function is discussed in detail. Finally, certain pitfalls related to in vitro slice transport studies are presented.


Differential effects of nephrotoxic agents on renal transport and metabolism by use of in vitro techniques.

A number of studies by the author and other investigators are reviewed in which the in vitro kidney slice technique has been used to evaluate the nephrotoxicity of various compounds. The kidney slice technique can be used to determine the effect of prior drug treatment of laboratory animals on renal organic acid (p-aminohippurate) or organic base (N-methylnicotinamide) transport, on glucose synthesis, and on oxygen consumption by renal coritical slices. The nephrotoxic agents uranyl nitrate and potassium dichromate exert inhibitory effects on renal function, althouhg both agents enhance organic base transport at low doses and potassium dichromate enhances organic acid transport at moderate doses. Enhanced PAH transport has been found to be a sensitive indicator of gentamicin induced nephrotoxicity, while inhibition of other parameters has been reported. The tissue slice method is less effective in evaluation chronic nephrotoxicity such as that produced by lead. The inhibitory effect of mercurial diuretics has been shown to be due to the general depression of metabolic activity by mercury. The kidney slice technique has been found to be a sensitive indicator in the assessment of halogenated hydrocarbon-induced nephrotoxicity. Differential effects of compounds on in vitro organic acid and base trasport provides information about the transport of these compounds as well as about their nephrotoxicity. Although it is often desirable to perform in vivo tests or other in vitro renal function tests, the kidney slice technique has proved to be extremely useful in toxicological evaluations.


Propagated responses produced by pacemaker stimuli falling within the QRS complex.

In a patient with idiopathic hypertrophic subaortic stenosis, propagated responses followed testing stimuli (St2) delivered within the wide QRS complexes elicited by driving stimuli (St1). This phenomenon was not seen at all St1-St2 which produced the minimal V1-V2 interval. It appeared at St1-St2 intervals which resulted in the largest difference between St1-St2 intervals, probably because of the marked asymmetry of propagation occurring very early in the cycle. More studies are required to determine whether this was due to a shift of the vulnerable period toward early systole (suigenesis to idiopathic hypertrophic subaortic stenosis) or to more complex types of intraventricular reentry. In other clinical conditions a similar mechanism could explain the multiple responses produced by pacemaker stimuli falling at the end of ventricular depolarization and also by "properly synchronized' low intensity transthoracic electrical discharges.


Generation of cytotoxic cells in absence of blastogenesis by mouse leukemic cells in mixed cultures.

Primary virus-induced murine leukemias and transplantable leukemic cell lines, originally induced by virus, chemical or physical agents, failed to stimulate normal spleen cells differing at the H-2 complex or at Mls locus in the one-way mixed leukocyte culture. Lack of stimulation seems at least partially due to a nonspecific inhibitory effect since in a "three-party" culture system the leukemic cells interfered with the mixed leukocyte reaction (MLR) of normal allogeneic cells. Characterization of the leukemic cell populations used, according to their T or B cell origin, disclosed that in most cases T cell markers were present. Also, no Ia antigens were detected on leukemic cells. Furthermore, using a 51Cr release assay, leukemic cells could sensitize allogeneic normal lymphocytes to kill specifically normal as well as leukemic targets, even when no MLR was detected in parallel cultures.


Absence of the gamma-spindle loop in the reinnervated hind leg muscles of the cat: "alpha-muscle".

Degeneration of afferent and efferent fibres supplying leg muscles was elicited by applying dry ice to the sciatic nerve in the mid femoral region. Two months after the denervation the cats showed practically normal walking activity. After 3 months the twitch tension of the reinnervated muslce (triceps surae) was almost similar to the tension developed by the non-operated contralateral control muscle. Afferent discharges with a pattern typical for muscle spindles could rarely be found before the end of the 4th month. The muscle spindles which could be recorded before the end of the 6th month showed mainly phasic responses to ramp-and-hold stretches and only a few of them exhibited a pause during muscle contraction. None of them showed any sign of gamma innervation, as tested by several independent  gamma-activating or gamma-blocking precedures. The conduction velocities of the regenerated afferent and effenent fibres were lower than those of the normal fibres; they gradually recovered with time after the operation but did not reach normal values by the end of the 6th month. Fibre histograms revealed that the afferent fibres of groups I, II and III had regenerated, while of the efferent fibres only those corresponding to the alpha group had regenerated. Since during the period of observation the reinnervated muscles lack the support of a functioning gamma innervation, it is proposed to call them "alpha muscles".


Details of glycosaminoglycan conformations and intermolecular interactions.

Among the mucopolysaccharides, similarities of chemical constitution are manifested in the similar molecular conformations that can be assumed by most of them. The differences, however, result in a preference for particular conformations. Small amounts of one glycosaminoglycan in a mixture can change the conformational preference of the major component. Advances in the techniques of fiber diffraction analysis allow us to visualize the roles of water and ions in some of these structures and to rationalize the strikingly different effects produced by Ca2+ and Na+ in hyaluronate systems, for example.


The effects of fasting and streptozotocin diabetes on hepatic triglyceride lipase activity in the rat.

The activity of hepatic triglyceride lipase in the rat was reduced by fasting. Withdrawal of insulin from insulin-treated streptozotocin-diabetic rats resulted in a decrease in hepatic triglyceride lipase activity. The behavior of the enzyme in both situations was similar to that of adipose tissue lipoprotein lipase. It is concluded that hepatic triglyceride lipase, like adipose tissue lipoprotein lipase, is under hormonal regulation by insulin.


Axonal dwindling in early experimental diabetes. I. A study of cross sectioned nerves.

A diminution of the cross sectional area of myelinated fibres in the common peroneal nerve was observed in rats four weeks after the induction of diabetes with streptozotocin. Small fibres were affected more than larger ones and the axon reduction was twice that of the myelin sheath. The fibre diminution and the decreased axon/myelin ratio may explain the slowing of nerve conduction in experimental diabetes.


Axonal dwindling in early experimental diabetes. II. A study of isolated nerve fibres.

In a morphometric study of isolated fibres of the common peroneal nerve in short-term diabetic rats reduced fibre calibre was observed. No segmental demyelination or remyelination was found, but the nodes of Ranvier were slightly widened and paronodal bulbi were swollen relative to fibre calibre. It is suggested that axonal dwindling is the primary event in experimental diabetes. The reduction of the myelin sheath may be a consequence of the abnormal nerve cell offshoot. The results obtained suggest that streptozotocin diabetes in the rat is a useful model for the elucidation of diabetic neuropathy.


Proliferative response of T lymphocytes to allogeneic tumor without generation of killer T lymphocytes in the presence of Ehrlich ascites tumor fluid.

The presence of cell-free Ehrlich ascites tumor fluid in an appropriate concentration was shown to permit a proliferative response, strictly dependent on the presence of Thy-1-positive lymphocytes, to proceed when introduced on the initiation of mixed lymphocyte-tumor reaction of C57BL/6 lymphocytes and Mitomycin-C-treated C3H/He myeloma cells, while it clearly inhibited the development of cytotoxic killer T lymphocytes toward the priming allogeic tumor cells. The fluid had no effect, however, when added to normally primed spleen cells at the onset of cytotoxicity test. Further investigation of the mechanism in vitro by ascitic tumor fluid selectively suppresses the generation of killer T lymphocyte activity to allogeneic target cells may lead to a better understanding of the suppressed T-cell responses observed in Ehrlich tumor-bearing animals, and additionally, may provide an intriguing information about the interactions between the distinct T-cell subsets responsible for proliferative response and generation of killer T lymphocytes.


Caffeine potentiation on lethality of L-1210 cells treated with neocarzinostatin.

The caffeine potentiation on lethality of L-1210 cells treated with Neocarzinostatin was studied. Colony formation of L-1210 cells treated with Neocarzinostatin was significantly decreased by incubation of the cells for 48 hr with 0.5 mM of caffeine which alone did not affect the growth rate and colony-forming ability of the cells. These results indicate that Neocarzinostatin may induce repairable damage in DNA of L-1210 cells.


Regulation of phosphate metabolism in Neurospora crassa: identification of the structural gene for repressible alkaline phosphatase.

Five additional mutants of Neurospora crassa have been isolated that lack the repressible alkaline phosphatase. The mutations in these strains map at a previously assigned locus on Linkage Group V designated pho-2 (GLEASON and METZENBERG 1974). The five new mutants, as well as three previously isolated by GLEASON and METZENBERG (1974), were examined for the presence of cross-reacting material to antibody prepared against purified wild-type enzyme. Two of the mutants produced high levels of cross-reacting material, thus providing evidence that the pho-2 locus includes the structural gene for the repressible alkaline phosphatase. Two revertants were obtained from one of the mutants that contained cross-reacting material. Neither revertant produced an enzyme that could be distinguished physicochemically from that of wild type. A method for measuring very low levels of repressible alkaline phosphatase in crude extracts is also described.


Regulation of phosphate metabolism in Neurospora crassa: identification of the structural gene for repressible acid phosphatase.

A mutant of Neurospora crassa with an altered repressible acid phosphatase has been isolated. The enzyme is much more thermolabile than that of wild type, and has an increased Michaelis constant. Tests of allelic interactions (in partial diploids) and in vitro mixing experiments were consistent with the mutation being in the structural gene for the enzyme. This gene, pho-3, was found to be located in the right arm of Linkage Group IV (LGIV). Thus, pho-3 and the structural gene for repressible alkaline phosphatase, pho-2 (LG V), map in separate linkage groups and cannot be part of the same operon. Neither of these structural genes is linked to the known regulatory genes, nuc-1 (LG I), nuc-2 (LG II), and preg (LG II).


[Incidence and manifestations of the heterozygosity of the gene causing diabetes].

A model of intermediate inheritance of diabetes mellitus is given, according to which juvenile diabetes developes in persons homozygous for the mutant gene, and a part of heterozygous carrier of the mutant gene has late diabetes mellitus. A large group of diabetes cases (about 50%), which are not due to direct diabetes heredity, is taken into account. Calculations are carried out on the basis of Hardy-Weinberg law from the patient questionaire data. The frequency of juvenile diabetes (developing within first thirty years) is 0,028% in the Leningrad population, the frequency of heterozygous diabetes gene carrier being 3,28%. Penetrancy of the diabetes gene in heterozygous state, as estimated from registrable (clinically expressed) forms of the disease, is 8,4%. The risk of juvenile diabetes in children, if one of the parents has juvenile diabetes, is 1,65% (3,3%-1/2), the frequency of heterozygous carriers in the population being 3,3%. The analysis of questionaire data of diabetes patients produced results, which were close to expected and indirectly confirmed the accurancy of the model proposed.


[Genetic analysis of microcephaly].

A clinical genetic examination of 22 cases of hereditary microcephaly in 11 families was carried out. Parent consanguinity (first and second cousins) was ascertained in 7 families. Genetic analysis of the experimental material performed by Weinberg's method showed significantly recessive course of the genuine microcephaly inheritance. High frequency of low degree mental retardation (30%) among probable recessive genes carriers of microcephaly (of the parents and their siblings) was ascertained and that confirmed the possibility of gene heterozygous state manifestation. The gene frequency fluctuated from 0,0042 to 0,024. About 3500 people might show low forms of mental retardation in connection with probable non-penetration (about 30%) of the microcephaly genes being in heterozygols state.


[Viability of beta-thalassemia hetero- and homozygotes in several populations of Central Asia].

Viability of hetero- and homozygotes for beta-thalassemia was studied in two isolated populations born after 1950 in conditions of malaria absence. Pregnancy outcomes and probability to get 16 years old were compared in marriages, in which one or both parents were heterozygous for beta-thalassemia, and in marriages, with both parents being normal. The ratio of children with normal genotype and children heterozygous for beta-thalassemia in families, where one parent was heterozygous for beta-thalassemia. Preliminary conclusions are made on the basis of the data obtained. Viability of children in families where one parent is heterozygous for beta-thalassemia is found to decrease in the absence of selective factors (malaria) as compared with normal children. The highest viability decrease was observed in the progeny of those families where both parents were heterozygous beta-thalassemia. Elimination mainly takes place within the first year of life.


Cholangiography in the jaundiced patient.

Though local practice will reflect the previously acquired expertise of the operators, it seems reasonable to employ a minimum of percutaneous transhepatic cholangiography, and, ideally, this in combination with ERCP for preoperative cholangiography in patients with cholestatic jaundice. Few cases will defy both techniques. The morbidity is well known and if properly anticipated can be reduced to a minimum by judicious use of antibacterial agents and early surgical intervention when appropriate. Grey-scale ultrasonography by indicating the diameter of the bile ducts enables one to select percutaneous transhepatic cholangiography for dilated ducts and ERCP for non-dilated ducts with an almost 100% success rate for the former and only slightly less for the latter in experienced hands.


[Congenital diaphyseal venous dysplasia (author's transl)].

A condition is described in which there is congenital ectasia of the veins in the lower extremities, in the tibia as well as in the soft tissues of the leg. The most striking symptom is bone pain, and the soft tissues are involved only to a limited degree. The condition has therefore been called "congenital diaphyseal venous dysplasia".


Landmarks of the knee joint of the lateral radiograph during rotation.

On specimens of human knee joints a study was made of the contours on the lateral radiograph during rotation up to 45 degrees inwards and outwards. The most important characteristics of the femoral condyles are the situation of the limiting groove on the condylar joint surface. The best for identifying the tibial condyles is the lateral intercondylar tubercle which proceeds backwards in an even, convex arch continuing direct in the posterior contour of the lateral tibial condyle.


[An analysis of the radiological appearance of osteoplastic metastases and osteitis deformans (Paget's disease) (author's transl)].

The validity of individual radiological signs for the differentiation of osteoplastic bone metastases from osteitis deformans has been investigated quantitatively (103 skeletal metastases from carcinoma of the prostate, 45 cases of Paget's disease, two osteoplastic bone metastases from a carcinoma of rectum and bronchus). The similarities were demonstrated by three cases observed by us. Problems in the quantitative evaluation of the radiographs are discussed.


[The radiological and scintigraphic findings in psoriatic arthropathy (author's transl)].

The radiological and scintigraphic findings in the skeletons of patients with psoriasis and psoriatic arthropathy are described. The results indicate that is it possible to detect inflammatory disease in joints by scintigraphy in patients who have not yet developed symptoms. The effect of treatment can be satisfactorily followed by serial scans. Further investigations and observations will be required to show whether the paravertebral calcification and syndesmophyte formation which has been described is really pathognomonic for psoriatic arthropathy.


[Congenital bowing of the tibia in neurofibromatosis von Recklinghausen (author's transl)].

There is a close, but not constant, relationship between congenital bowing of the tibia or infantile pseudo-arthrosis of the tibia and von Recklinghausen's neurofibromatosis. In about 40% it is possible to confirm the diagnosis by examining relatives, but the radiological findings and the course of the disease, as well as by histological examination. Anterior bowing with dysplastic-sclerotic bone changes results in the "high risk tibia". This may lead to a pathological fracture. Treatment depends on the clinical findings and radiological changes. It should be aimed at preventing pathological fractures and pseudarthrosis.


[Cerebral anatomy on computerised tomograms (author's transl)].

An attempt has been made to show which anatomical structures of the skull and brain can be demonstrated by analysing 668 computerised tomograms of normal brains. Many parts of the bony skull, the cisterns and ventricles and physiological calcification of the pineal body, choroid plexus and falx can be clearly identified. Frequently it is possible to distinguish between grey and white matter in the basal ganglia. After intravenous contrast injection it is sometimes possible to identify arteries and other pathways and vascular structures by their increased contrast. Because of the constant topographic relations in the brain, it is possible to identify these structures with certainty on the computerised tomogram.


[Angio-tomographic aspects of cerebral aneurysms and angiomas (author's transl)].

Angio-tomography is a selective radiological method which can be used to complement serial angiographs in order to demonstrate better detail. This results from the fact that selective levels can be demonstrated without super-imposition of bone or other vessels. Angio-tomography combines the advantages of tomography, subtraction and selective procedures. The method has been used for all types of intracranial disease; aneurysms and angiomas are the most common conditions for which angio-tomography has been employed. The present paper is based on a total of 60 cerebral aneurysms and angiomas in various situations; angiotomography was indicated for these amongst a larger material of intracranial vascular malformations in which the diagnosis was possible without its use. The indications consist of: 1) Demonstration of very small aneurysms and their differentiation from vessel loops, 2) Demonstration of the origin of an aneurysm which is situated at the confluence of several major vessels, 3) Point of origin of a so-called hugeaneurysms which cannot otherwise be shown by oblique views, 4) Demonstration of so-called micro-angiomas and of the afferent and efferent vessels in central arterio-venous malformations. Unusual aneurysms of the internal carotid artery, of the anterior choroidal and vertebral arteries are described. Some new concepts in the localisation of aneurysms of the anterior communicating artery are discussed. Many of the examinations were carried out with an angio-tomographic apparatus made in the department.


[The diagnosis of cerebellopontine tumours by tomography after positive contrast cisternography (author's transl)].

Forty-one tumours in the cerebello-pontine angle were found as a result of 207 Duroliopaque cisternograms. In all 37 patients who came to operation, the position and size of the tumour, as shown radiologically, could be confirmed. The value of positive contrast cisternography was greatest in the smaller tumours. 35 tumours were less than 2.5 cm. in their vertical diameter. Twenty-four tumours were predominantly extra-meatal, were predominantly extra-meatal, four predominantly intra-meatal and 13 extra- and intra-meatal as well. In only 23 patients was there enlargement of the internal acoustic meatus, either absolutely or by comparison with the other side. In 18 patients there was no significant difference in the meati on the two sides. These facts and the observation that there may be idiopathic meatal widening in the presence of typical symptoms of an acoustic neuroma, but without a tumour being present, underlines the importance of positive contrast cisternography. A high degree of accuracy for tumour less than 2.5 cm. can only be obtained by using the method in conjunction with multi-dimensional tomography.


[A new film changer for use with an image intensifier (author's transl)].

A new film changer is described which permits serial films to be obtained, it can be used without attenuation of radiation to the image intensifier. Details of the serial changer, such as speed and resolution are described.


[Evaluation of experimental arterial occlusion, with special reference to its complications (micro-spheres, Histo-Acryl, Paladur, alternating and direct currents (author's transl)].

Permanent arterial occlusion can be obtained experimentally by using plastic micro-spheres, polymerising materials (Histo-Acryl, Paladur) and direct current thrombosis. Certain limitations in their use arise from the properties of the substances. In our experience, plastic microspheres are not suitable for blocking entire organs or large vascular territories. They are more appropriate for occlusion of individual, small vessels by super-selective injections. Sole use of polymerising materials is not possible; they are best used in combination with other techniques (magnets, balloon catheters). Induction of thrombosis by small direct currents (50 volt, 10 mA). produces permanent and complete occlusion. The method can be used selectively or super-selectively. Unwanted thrombosis can be avoided by using catheters with the smallest possible external diameters.


[The application of sonography in the differential diagnosis of space-occupying renal lesions (author's transl)].

Two dimensional ultrasound echography is a reliable method for demonstrating the topography of retroperitoneal space-occupoing lesions which causes little inconvenience to the patient. This is particularly the case in the upper abdomen, where the differentiation between renal, suprarenal, pancreatic and splenic lesions has been difficult. The criteria for the differential diagnosis of sonographic findings are described and discussed. Echography should be used before invasive methods, particularly angiography, are employed.


[Urographic investigations on the effect of high dose progestagen therapy (author's transl)].

The relationship between high dose progestagen treatment and changes on the intravenous urogram were studied in 27 patients. For this purpose, six specific features in the urogram were examined. Our findings in the patients available to use indicate that there is no demonstrable dilatation of the renal pelves and ureters as a result of the administration of progestational substances alone. These results are discussed in relation to the relevant literature.


[Film quality in film mammography. Article II: Signal/noise ratio of the recording system as a quantitative measure of quality. Final evaluation of recording systems (author's transl)].

During consideration of three film mammographic systems (see article I), the concept of signal/noise ratio is developed as a quantitative measure of film quality. The ability to recognise detail related to detail size, film blackening and exposure geometry was studied for various systems, and the quality profiles are discussed. There is a considerable difference in quality between industrial films without screens and film-screen combinations; however, exposure geometry during mammography has a considerable effect which tends to reduce the difference. Consequently, detail sizes of 200 mu to 1,000 mu (including the majority of mammographic micro-calcifications) are shown about equally well. Contrast for the lo-dose system (D = 1.1) is somewhat less than for adequately exposed industrial film (D = 2.0) or for Cronex 75 m with lo-dose screen (D = 1.5 to 2.0). The lo-dose system (D = 1.1) is 8.6 times more sensitive than Cronex 75 m (D = 2.0), and the system Cronex 75 m with lo-dose screens is 11.6 times more sensitive. Over-exposure with the lo-dose system, contrary to industrial film, rapidly leads to unsatisfactory results (loss of contrast and increased granularity for D greater than 1.5). On the other hand it is often not possible to obtain an adequate exposure (D = 2.0) when using industrial film. For these reasons it is often an advantage to examine large breasts (with considerable geometric unsharpness) and the dense breasts of young women with a film-screen combination (eg. lo-dose system) which requires approximately one eighth of the dose necessary for industrial film. For small or easily compressable breasts (little geometric unsharpness) best results are obtained, using an adequate exposure by employing industrial film; radiation dose it then acceptable. Detailed analysis of the systems has shown that the recording system unsharpness of film-screen combinations in mammography should be reduced.


Some effects of experimentally-induced diabetes on pituitary-testicular relationships in rats.

The effect of experimentally-induced diabetes mellitus on reproductive organ weights, serum and pituitary gonadotropin levels and serum testosterone levels was studied in 3-month old rats. In experiment 1, intact rats were treated with alloxan monohydrate or streptozotocin. In experiments 2 and 3, intact and castrated rats were rendered diabetic with alloxan (experiment 2) or streptozotocin (experiment 3). The duration of each experiment was 3 weeks. In each experiment diabetes resulted in body weight losses or reduced body weight gain, elevated serum glucose concentrations and reduced assessory sex gland weights (intact rats). Serum levels of testosterone were depressed (P less than 0.05 or P less than 0.01) in diabetic rats. Serum levels of LH were significantly (P less than 0.05) lower in intact diabetics than in controls when pooled data from the three experiments were compared. Serum levels of FSH were not affected by diabetes. Pituitary concentrations of FSH were elevated (P less than 0.05) in diabetics in two of the three experiments, while LH concentrations were elevated (P less than 0.05 or P less than 0.01) in diabetics in all experiments. The hypersecretion of gonadotropins in castrated rats was not affected by diabetes.


Concentration and adenosinetriphosphatase activity on left ventricular actomyosin in Goldblatt rats during the compensatory stage of hypertrophy.

Left ventricular myocardia of Goldblatt rats with an average increase in arterial blood pressure to about 200 mm Hg showed a progressive reduction of the Ca-activated specific acotmyosin ATPase activity 4 -12 weeks after the coarctation of one renal artery, as compared with controls of the same age. During the same period, a significant increase in the concentration of contractile proteins was noticeable, whereas the content of nonprotein substances and of water corresponded to the control values. The hydroxyproline concentration, as a measure of the collagen tissue content, increased only after 24 weeks. The time course of the specific ATPase activity was closely parallel to the decrease in the unloaded myocardial shortening velocity, as estimated at the same stage by our group. This is in accordance with the assumption of a fundamental relationship between the two values. The reduced rate of energy turnover and of the shortening velocity is regarded as an adaptive mechanism which, however, has a negative effect in advanced hypertrophy when further diminution takes place. The decrease in the specific enzymatic activity of actomysin is not necessarily linked to a large increase in myocardial mass, but is already apparent at moderate degrees of hypertrophy (34%).


Mitochondrial adenosine triphosphatase from yeast, Saccharomyces cerevisiae. Purification, subunit structure and kinetics.

1. A procedure for the purification of ATPase extracted by chloroform from baker's yeast (Saccharomyces cerevisiae) is reported. The yield based on submitochondrial particles was 55% and the purification was 100-fold. The isolated complex was homogenous as assessed by gel filtration, ion-exchange chromatography, sedimentation in sucrose gradient and in the analytical ultracentrifuge. The molecular weight determined by gel filtration was 400000 +/- 20000. Ultracentrifugation yielded s020,w = 12.50 +/- 0.13 S and the laser light scattering study gave a diffusion coeficient of D20w - 2.92 X 10(-7) cm2 s-1. The amino acid composition as well as absorption, fluorescence, and circular dichroism spectra, from which the helicity of 39% was evaluated, are given. 2. On polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate, six components with molecular weights of 58500(alpha), 55000 (beta), 42000, 34000 (gamma), 10000(delta), and 8600 (epsilon) were observed with a stoichiometry of 3:3:1:1:1:1. The amino acid composition is given for alpha + beta and gamma as well as delta and epsilon components. 3. The maximum specific activity of the enzyme was 200 U/mg under the optimum conditions. The enzyme was inactivated by incubation at 0 degrees C and strongly inhibited by the antibiotic Dio-9 but not by oligomycin and N, N'-dicyclohexyl-carbodiimide. The effects of kinetic parameters and anions on the enzyme are reported. Two active sites for Mg-ATP with Km values of 0.045mM and 0.37mM and a single activie site for Mg-ITP with Km = 0.179mM were found. A study of the temperature dependence of the maximum activity revealed a straight line in the Arrhenius plots with an activation energy of 11.0 kcal/mol (= 46 kH/mol).


Lack of linkage between gene(s) controlling the synthesis of the seventh component of complement and the HLA region on chromosome No. 6 in man.

The family of an individual was studied who lacks the seventh component of complement in his serum (C7 homozygous deficiency). Both parents are C7 heterozygous-deficient. In this investigation, the following parameters were determined: complement components in functional and immunochemical tests; HLA-A,B antigens, HLA-D (MLC) determinants; the Bf system; glyoxalase I and B cell antigens. No evidence for linkage between the immunogenetic linkage group on chromosome 6 and gene(s) controlling the synthesis of the seventh component of complement was obtained. This is in according with the assumption that only genes controlling components of the initiating rather than the membrane attack unit of complement are linked to the HLA region.


Mitosis of maternal lymphocytes in the presence of fetal cells: possible implication in prenatal diagnosis from fetal blood samples.

The suppression of proliferation of maternal lymphocytes by the lymphocytes of their own male newborns have been tested in a PHA-induced two-way stimulation system. The mixed lymphocyte cultures of 6 out of 12 such mothers/son pairs had 23--50% metaphases with 46,XX karyotype. In 2 more cases 10% maternal metaphases have been observed. Hence, it appears that fetal lymphocytes are unable to suppress the proliferation of maternal cells completely.


Genetic loci of components of the classical and alternate pathway of complement activation: a new dimension of the immunogenetic linkage group (HLA) on chromosome 6 in man.

In this review, a working hypothesis is put forward that functional cooperation of various types of cells and proteins in immune recognition, mediation and response is maintained by a common chromosomal region which evolved over millions of years from a common ancestor by gene duplication. In brief, the known functions of the H-2 complex are discussed (susceptibility and resistance to viral infection, immune response genes, T-B cell interaction as non-self recognition and response). The addition of loci of the classical and alternate pathway of complement activation to the HLA region (i.e., C2, C4 and the Bf system) is reviewed with respect to functional relationship to immune recognition and mediation mechanisms. As expected according to this hypothesis, genes for late-acting components (C3, C5, C7 and C8 in man, C5 in mice) have so far not proved to be linked to HLA.


Lymphocyte transformation in human plasma without addition of synthetic medium. A study of immune function in patients with chronic uraemia or diabetes mellitus.

The in vitro response of lymphocytes obtained from normal subjects, uraemic patients on haemodialysis and diabetic patients was studied using cultures containing either medium plus plasma (medium cultures) or plasma alone (plasma cultures). The study demonstrated that plasma alone can adequately support lymphocyte transformation induced by nonspecific mitogens (PHA and PWM) and allogeneic lymphocytes in mixed lymphocyte culture (MLC) reaction. This investigation further confirms our previously reported findings that uraemic patients undergoing haemodialysis have a normal lymphocyte response in MLC and to PHA and PWM. Plasma cultures give results similar to conventional medium cultures in subjects where lymphocyte transformation is normal. The lymphocyte hyporactivity observed in diabetics is, however, better shown in the plasma cultures. The suppressed response of the diabetic patient's lymphocytes to PHA and PWM both in the presence of autologous and normal AB plasma suggests intrinsic lymphocyte dysfunction as the explanation for impaired immune function. Plasma cultures may provide a better in vitro system for the evaluation of immune function in certain groups of patients where it is desirable to distinguish between intrinsic abnormalities of lymphocyte function and the effect of humoral immunosuppressive factors.


[(Whole body plethysmographic studies on the bronchospasmolytic effect of injectabls drugs. Comparison of aminophylline, diprophylline and hexoprenaline)].

The bronchodilator effect of intravenously administered drugs was tested by whole bodyplethysmography in 24 patients with chronic obstructive airway disease over a period of 60 min. Subsequent to this, another bronchodilator was applied by a metered dose inhaler. Aminophylline (0,24 g i.v.) was followed by a decrease of airway resistance to a maximum of 73,04% of the base line value after 30 min. and to 76,33% after 60 min; inhalation of fenoterol (0,4 mg) caused a further decrease to 47,33% after 20 min. Diprophylline (0,28 gi.v.) produced only a slight decrease to a maximum of 95,75% after 20 min. and to 99,54% after 60 min.; fenoterol (0,4mg) reduced airway resistance to 55,29%. Hexoprenaline (0,005 mg i.v.) was followed by a decrease to a maximum of 74,71% after 5 min. and to 92,83% after 60 min.; ipratropiumbromide (0,08 mg) caused a further decrease to 54,92%. A considerable increase of heart rate, from 89,25/min. to 106,15/min. occurred only after hexoprenaline. For intravenous bronchodilator therapy we prefer aminophylline. Optimal bronchodilation however, was seen only after further application of fenoterol or ipratropiumbromide by a metered dose inhaler.


Comparison of the results of semi-thin sections with those of conventional paraffin histology and immunohistology in renal biopsy specimens.

One hundred renal biopsy specimens out of the routine material were investigated by the usual paraffin technique, by semi-thin sections and by the immunofluorescence technique. The findings were compared in regard of the diagnoses made by each technique. The results indicate that the paraffin technique is inadequate in a considerable proportion of the specimens, because very small glomerular lesions cannot be recognized. The semi-thin section technique enabled us to correct the light-microscopic diagnosis in 17 per cent, particularly in establishing membranous glomerulonephritis. In another 31 per cent the histologic diagnoses could be improved. The immunofluorescence technique provided evidence of active glomerulonephritis in 57 per cent (24 cases with immune complex nephritis and 3 cases with basement membrane antibody nephritis). We found a much closer correlation between results of the semi-thin section technique and immunohistology that between the paraffin technique and immunohistology.


Autoradiographic detection of mucopolysaccharide accumulation in single fibroblasts.

A method is described for localizing acid mucopolysaccharides autoradiographically in cultured cells. Normal fibroblasts and fibroblasts, from patients suffering from Mucopolysaccharidosis II disease (MPS II), were cultured for six days in the presence of 35SO4 and one day in unlabelled medium. The cultured cells were transferred to plastic film dish and, after settling, they were rapidly quenched, freeze-dried, fixed in osmium tetroxide vapour and embedded in Epon. Grain counting after autoradiography in 2 mum sections revealed a significant difference (P greater than 0.001) in 35SO4 incorporation in the perinuclear cytoplasm of MPS II cells and control cells grown under the same conditions. Autoradiography was also performed after mixing MPS II cells and control fibroblasts in a ratio 1:1-8 prior to freezing and the same ratio was found between labelled and unlabelled fibroblasts. These results demonstrate the feasibility of the present autoradiographic technique for the detection of the acid mucopolysaccharide storage at the single cell level.


Histochemical sub-types of three fibre-types of avian skeletal muscle.

The soleus muscle of adult chicken was studied histochemically. Succinic dehydrogenase (SDH) and myofibrillar ATPase reactions, with or without preincubation in K2-EDTA salt, were compared in serial frozen sections. Based upon the distributions of the above reactions, the three major fibre types distinguished were "Type I red", "Type II red" and "Type II white". On the basis of non preincubated ATPase reaction alone two sub-types of type I red fibres could be distinguished. However, following preincubation in a "Cold" solution of K2-EDTA, Type II red fibres fell into two sub-types and Type II white fibres fell into three sub-types. Amalgamating the two already existing classifications, a more elaborate classification is presented for characterizing these different sub-types. The presence of two different or a spectrum of staining variations in a seemingly homogeneous population of muscle fibres in a given fibre-type emphasizes the possible correlation between this histochemical data and the heterogeneity of contraction times of the different motor units.


Implications of the normalization principle for psychotic children.

The evolution of the normalization principle is described. Whereas the principle initially advocated normalizing individual lives and routines, it presently is being applied to entire human service delivery systems. The need for more data supporting proposals to normalize human service systems is discussed, as well as some disadvantages in applying this approach with psychotic children. It is concluded that a wide range of treatment options for psychotic children is needed and that these options should be evaluated based on their effectiveness with individual children.


Characterization of qa-2 mutants of Neurospora crassa by genetic, enzymatic, and immunological techniques.

Genetic and complementation mapping studies using 20 qa-2 mutants defective for catabolic dehydroquinase indicate that the qa-2 gene encodes a single polypeptide chain and is the structural gene for catabolic dehydroquinase, a 220,000-molecular-weight protein composed of identical 10,000-molecular-weight subunits. Many qa-2 mutants are capable of reversion, but no evidence has yet been obtained for nonsense mutations in this gene. The biochemical consequences of the mutations in two complementing qa-2 strains (M239 and M204) have been determined. Both mutants have extremely low levels of catalytic activity and form a heterocaryon with about 4% of the wild-type activity. As assayed by immunological cross-reactivity, mutant M239 and the heterocaryon have nearly wild-type levels of native-molecular-weight catabolic dehydroquinase protein, whereas M204 has no detectable amount of this protein. Thus it is concluded that M239 has a mutation at or near the catalytic site which reduces the activity 10,000-fold but has little or no influence on the formation of the native multimeric structure. In contrast, M204 apparently has a mutation that severely inhibits aggregation and may have only a minor effect on the inherent potential for catalytic conversion at the reactive site. The heterocaryon would appear to form a mixed multimer with the monomeric subunits from M239 providing the aggregated structure and those from M204, the catalytically active moiety.


Mutations in prophage phi11 that impair the transducibility of their Staphylococcus aureus lysogens for methicillin resistance.

Methicillin resistance (mec) is not transduced into Staphylococcus aureus 8325-4, but is transduced into this host after it has been lysogenized with phage phi11 and has acquired the penicillinase plasmid pI524 by a separate transduction (Cohen and Sweeney, 1970, 1973). Strain 8325-4 is competent for transformation of typical plasmid or chromosomal markers and for mec only if it is lysogenic for phi11 or a related prophage (Sjöström et al., 1974, 1975). A mutant strain of phi11 that was temperature sensitive (Ts) for vegetative multiplication did not mediate competence for transformation of its 8325-4 lysogen if the lysogen had been grown at a nonpermissive temperature (Sjöström and Philipson, 1974). We isolated four Ts mutants of phi11 that did not mediate transducibility of their 8325-4(pI524) lysogens for mec after growth at nonpermissive temperatures (40 to 42 degrees C). Transduction of typical plasmid or chromosomal markers was not affected. These phi11-Ts mutants mediated normal competence of their lysogens for transformation of a tetracycline resistance plasmid. Similarly, phi11-Ts mutants that rendered their lysogens temperature sensitive for transformation did not depress the frequency of transduction of mec. These two types of phi11-Ts mutants fell into two different genetic complementation groups that differed in the physiology of deoxyribonucleic acid synthesis and in the time of expression of the mutations during a single-burst growth cycle at a nonpermissive temperature. A virulent mutant of phi11, which plaqued with 100% efficiency on 8325(phi11), also failed to condition strain 8325-4 for transducibility of mec but retained the ability to confer competence for transformation of a tetracycline resistance plasmid. Different genetic loci and physiological functions are involved in phi11 mutations that affect transducibility of mec and those that affect competence for transformation of markers generally in S. aureus 8325-4.


Biosynthesis of the outer membrane receptor for vitamin B12, E colicins, and bacteriophage BF23 by Escherichia coli: kinetics of phenotypic expression after the introduction of bfe+ and bfe alleles.

The bfe locus codes for the cell surface receptor for vitamin B12, the E colicins, and bacteriophage BF23 in the Escherichia coli outer membrane. When the bfe+ allele, which is closely linked to the argH locus, was introduced into an argH bfe recipient by conjugation, arg+ recombinant cells rapidly and simultaneously acquired sensitivity to colicin E3 and phage BF23. In the reciprocal experiment introducing bfe into an argH bfe+ recipient, it was found that colicin E3-resistant, arg+ cells began to appear shortly after the arg+ recombinant population began to divide. This was far earlier than would have been predicted on the basis of 220 receptors per haploid cell. Moreover, there was a lag between the appearance of colicin resistance and the appearance of resistance to killing by phage BF23, and hence a period of time during which some arg+ recombinant cells were sensitive to the phage but resistant to the colicin. Colicin E3 added to cells during this period of time protected against phage killing, indicating that the colicin-resistant cells still had receptors capable of binding colicin on their surface. The modification of the phenotypic expression of colicin and phage resistance by inhibitors of deoxyribonucleic acid, ribonucleic acid, and protein synthesis was also investigated. The results obtained indicate that the receptor protein coded for by the bfe locus can exist on the cell surface in several different functional states.


Interaction of cytoplasmic membrane and ribosomes in Escherichia coli: spectinomycin-induced disappearance of membrane protein I-19.

Incubation of Escherichia coli with spectinomycin caused the disappearance of a major protein from the cytoplasmic membrane. This protein, called "I-19", was not a ribosomal protein. Its disappearance was not a result of the direct action of spectinomycin on the cytoplasmic membrane, but a result of its action on ribosomes. The disappearance was specifically induced by spectinomycin, and other antibiotics such as neomycin, erythromycin, and chloramphenicol had no effect. Although growth was not required for spectinomycin-induced disappearance of protein I-19 from the cytoplasmic membrane, the disappearance was not observed under conditions where protein synthesis was inhibited completely either by the addition of chloramphenicol or by cooling in ice. It is suggested that at least some ribosomes interact with the cytoplasmic membrane and that a modification of the mode of interaction through the action of spectinomycin on ribosomes caused the deletion of membrane protein I-19.