[Effect of bacterial polysaccharide on the primary and secondary immunologic response following immunization with sheep erythrocytes]. The effect of typhoid bacterial polysaccharide on the primary and secondary immune response to SRBS was studied. The polysaccharide was shown to have both stimulating and depressive effect on the population of antibody-producing cells. This effect depended on the time and the number of polysaccharide injections. Thus, a single polysaccharide injection made on the day preceding immunization resulted in the maximum stimulation in the system of IgM- and IgG-producing cells, while the maximum depression of these cells could  be observed after 2 polysaccharide injections: on the day preceding immunization and on the day of immunization. In the secondary immune response considerable stimulation of the populations of antibody-producing cells was observed after polysaccharide injections made on days 2 and 3 after reimmunization.


[Effect of a typhoid lipopolysaccharide on the primary and secondary immunological response to ram erythrocytes]. The effect of typhoid bacterial lipopolysaccharide (LPS) on the primary and secondary response to sheep red blood cells was studied. LPS, injected simultaneously with the antigen, stimulated the synthesis of IgM and IgG, as well as the production of rosette-forming cells. When injected on days 2 and 3 after the secondary immunization, LPS induced the maximum stimulation of IgM, IgG and rosette-forming cells, while the injection of LPS prior to immunization induced immunosuppression which particularly affected IgG and rosette-forming cells.


[Characteristics of the immunosuppressive activity of lymphoid organ cells in the process of sensitization with ram erythrocytes and exposure to antilymphocyte serum]. In the adoptive transfer of cells obtained from the thymus, lymph nodes and the spleen to intact syngeneic animals the suppression of immune response was induced by lymph node cells. If the donors were previously sensitized, the cells of the thymus and lymph nodes showed suppressive activity in the adoptive transfer test. A single injection of antilymphocytic serum to the donors of lymphoid cells, previously sensitized with sheep red blood cells, enhanced the immunosuppressing action of thymocytes and lymph node cells.


Persisting suppression of prolactin secretion after long-term treatment with bromocriptine in patients with prolactinomas. The effect of bromocriptine withdrawal after long-term treatment on prolactin levels has been investigated in thirty-seven patients with prolactinomas. In ten patients with macroprolactinomas and post-operatively excessively high prolactin levels persisting suppression of prolactin secretion after bromocriptine withdrawal has been observed. This effect was not observed in patients with microprolactinomas or macroprolactinomas with only moderately elevated prolactin levels. The degree of persisting suppression correlated to the height of prolactin levels before treatment and to the duration of bromocriptine therapy. No correlation was found between the rise of prolactin levels after bromocriptine withdrawal and withdrawal time. It is suggested that the persisting suppression of prolactin levels is a sequence of reduction in tumour size. This anti-proliferative action of bromocriptine seems to be specific for the prolactin secreting cells in macroprolactinomas with high proliferation rate and high prolactin turn-over. These findings offer new possibilities in the management of patients with macroprolactinomas.


Interaction of synthetic progestagens with renal mineralocorticoid receptors. Progesterone increases urinary sodium excretion at least in part by competition for renal mineralocorticoid receptors. In contrast, synthetic progestagens do not increase sodium excretion or even cause a slight sodium retention. We therefore compared the ability of progesterone and 12 progesterone like compounds to displace [3H]aldosterone from binding at cytoplasmic mineralocorticoid receptors in rat kidney. All synthetic progesteronelike steroids were less active than progesterone in competing with [3H]aldosterone for the receptor binding: progesterone 100%, 11 beta-OH progesterone 50%, 17 alpha OH-progesterone 24% and 16 alpha-methyl-progesterone 3%. Derivates of 17 alpha OH-progesterone (medrogestone 5%, dydrogestone 4%, medroxy progesterone-acetate 2% and chlormadinone-acetate 0.5%) were more potent than 19-nor-testosterone derivates: ethisterone 1%, norethisterone less than 1%, norethindrone less than 1%, norethyl-nodrel less than 1%, and ethynodiol-diacetate less than 1%. The decreased affinity of synthetic progestins for mineralocorticoid receptors explains in part the lack of natriuretic activity of these compounds.


[Possibilities and limitations of echocardiography]. Echocardiography offers a noninvasive method ideal for the patient and the observer. The exact diagnosis can be established in most of cardiac diseases. We use the technique as a screening method to select patients for invasive procedures, it allows an accurate assignment and timing of catherization.


Immunoelectrophoresis in the diagnosis of neuroectodermal and mesodermal intracranial tumours, especially those of the posterior fossa. Immunoelectrophoresis of extracts of 200 intracranial tumours against rabbit anti-glioblastoma serum gave positive results (= precipitation) in all cases of tumours of neuroectodermal origin such as glioblastoma, astrocytoma, oligodendroglioma, ependymoma, neurinoma, and spongioblastoma. No immunoelectrophoretic precipitation was seen for any of the tumours of mesenchymal origin, for instance meningioma and metastases of cancer. On the basis of these findings, immunoelectrophoresis is considered to be a reliable method for differentiation between tumour tissue of neuroectodermal and non-neuroectodermal origin. Among the 41 posterior fossa tumours some unusual observations were made. Cerebellar angioblastoma (Lindau tumour) showed an atypically located precipitation line, which for the present is interpreted as an immunological reaction to vascular wall tissue. Furthermore, among the group of so-called medulloblastomas, two subgroups were distinguished on the basis of three parameters. The first of these subgroups comprises tumours whose immunoelectrophoretic pattern resembles that of gliomas, which are histologically characterized by neuroectodermal structures and which occur in younger children (5--10 years). The tumours of the second subgroup, which do not show this neuroectodermal immunoelectrophoretic pattern, have a sarcomatous character histologically, and occur in patients aged between 10 and 50 years. The view that medulloblastoma comprises a number of different types of tumour seems to be confirmed by this finding.


Generation and characterization of condensation aerosols of vanadium pentoxide and pyrene. Generation and characterization of submicron aerosols of V2O5 and pyrene, two materials with very different physicochemical properties, were studied. Vaporization-condensation methods were used to generate the aerosols. The effects of various experimental conditions on particle size and aerosol mass concentration were investigated. Also, chemical stability of the aerosols and the source materials with respect to temperature was determined. Experimental results show that there are significant differences in the nature of pyrene and V2O5 aerosols. These methods and experimental conditions can be easily adapted for generation of heterogeneous condensation aerosols of pyrene and V2O5.


Effect of oil mists on the irritancy of sulfur dioxide. II. Motor oil. This study examines the effect of sub-micrometer aerosols of motor oil on the irritant potency of sulfur dioxide. The increase in pulmonary flow resistance in guinea pigs was used as the bioassay of irritant response. When administered simultaneously, both unused and used motor oil protected against the irritant response to sulfur dioxide. This protective effect was lost when the oils had been previously reacted with sulfur dioxide. The additives used in the motor oil provided an equivalent protection when dissolved in a mineral oil which alone did not protect. Pre-exposure to motor oil did not provide protection. Sulfur dioxide and motor oil given together as a pre-exposure provided protection against further exposure to sulfur dioxide.


Christ-Siemens-Touraine syndrome--a clinical and genetic analysis of a large Brazilian kindred: II. Affected males. We describe 13 males with Christ-Siemens-Touraine syndrome from one family. History and examination were supplemented by three sweat tests and dermatolglyphic analysis. Some of the patients had two uncommon findings (onychodystrophy and excessive lacrimation), and five had an "incomplete" form of the syndrome. Four signs (distal phalanges of fingers and toes radially and tibially deviated, respectively; facial hypochromic spots; large occipitofrontal circumference) seem to be described for the first time. The segregation proportion in the sibships with at least an affected male was found to be normal (1:1) in 44 series of data (43 from the literature), where a high ascertainment bias was present (155 affected and 68 normal males).


Monosomy 21: a possible stepwise evolution of the karyotype. We describe a female infant with manifestations of complete monosomy for chromosome 21 intrauterine growth retardation, failure to thrive, craniofacial anomalies, arthrogryposis-like features, and psychomotor retardation. Chromosome analysis demonstrated mosaicism for three different cell lines in the various tissues examined; 45,XX,-21/46,XX,del(21)(q11)46,XX. The existence of these three lines suggests a possible explanation for the few cases of "complete monosomy 21" which have been reported.


Presence of a human chorionic gonadotropin--like substance in human sperm. An hCG-like material has been extracted from human sperm. These experiments were designed to characterize this material. Sperms of 10 volunteers were separated from seminal fluid, washed in PBS three times, and resuspended in 0.5 ml of the same buffer. Samples were pooled; cells were disrupted by sonication and extracted in alkaline buffer by constant agitation at 4 degrees C. The extract was ultracentrifuged at 4 degrees C. Supernate was lyophilized and reconstituted in 2 cc of distilled water. This material presented a dose-response curve parallel to those of IS2-hCG and CR119 in beta hCG RIA. When chromatographed in a Sephadex G-150 column the extract eluted within the hCG range and immunoreacted in the specific beta hCG RIA. When absorbed onto a concanavalin A--Sepharose column, all recovered immunoreactive material eluted after exposure to alpha-D-methylglucoside, indicating that it is a glycoprotein. The extract stimulated progesterone and testosterone secretion in porcine granulosa cells and decapsulated rat testis, respectively, indicating its biologic potency.


The effect of prenatal factors on crown dimensions. As shown in 870 white participants in the National Collaborative Perinatal Project (NCPP), maternal health status during pregnancy and birth size are systematically related to mesiodistal and buccolingual crown dimensions of I1, I2, dc, dm1, dm2 and M1. Maternal diabetes, maternal hypothyroidism and large size at birth are associated with larger maxillary and mandibular teeth in white children. Conversely, deciduous and permanent crown diameters are diminished in maternal hypertension, and in low birthweight and small birth-length conditions. These findings suggest that maternal and fetal (or gestational) determinants of both deciduous and permanent tooth crown dimensions may account for as much as half of crown-size variability with major implications to population comparisons and historical odontometric differences and trends.


Development of velvet coat (Ve/Ve), another early lethal mutation in the house mouse. A new semidominant mutation in the house mouse, velvet coat (Ve), is described. Ve homozygotes, recognizable on day 5 of gestation by their deficiency of ectodermal cells, never produce mesoderm and are resorbed by days 9-10. Primary Ve action may occur during the formation of the blastocyst or during determination and differentiation of the inner cell mass, or both. Accordingly, Ve/Ve embryos may provide a useful model for investigating primary gene action during blastocyst formation and subsequent differentiation.


Expertizing of posttraumatic sexual disturbances in males. An analysis has been performed of 95 expertises in men, who asked for compensation of after-effects in injury in their sex life. The regions most frequently affected were sexual organs, the head, the pelvis and the lower spine. Roughly one half of the examinees reacted strongly to the injury and its consequences. The most frequently reported symptoms were erectivity disturbances. The greatest subgroup consisted of men who had a rational attitude to an objective organic finding (about 38%). The second place is occupied by men in whom it was not possible to prove any somatic changes, but whose psychic reaction to injury was distinct and pronounced (about 31%). Roughly two out of three men reported a deterioration of their working capacity after injury and/or a disturbance of their interpersonal relations. Only 5% of the examinees underwent an expert examination during the first year following the injury. It is stressed that administrative procedures for damages should be performed with maximum speed especially in cases where there were no objectively proved after-effects.


Limitations of the cyanide electrode for use in plasma and whole blood. Although the cyanide ion-specific electrode has been used for the measurement of cyanide in biologic fluids, its validity has not been documented under these conditions. Therefore, we analyzed 60 paired samples of blood and plasma in a blinded fashion and compared the cyanide ion concentrations obtained from the ion-specific electrode with those obtained from a standard chemical assay. There was poor correlation between results in plasma samples (r = .52, n = 26) and no correlation in whole blood (r = .02, n = 34). Until refinements of the electrode technique are made, this method cannot be recommended for the measurement of cyanide in biologic fluids.


Psychomotor performance following exposure to trace concentrations of inhalation anesthetics. Using three psychomotor tasks administered three times each at 2-week intervals, we studied the performances of 18 control subjects and 18 subjects who were routinely and daily exposed to trace concentrations of anesthetic gases in the course of their clinical practice. No significant differences attributable to exposure to trace concentrations of anesthetics were detected. It is concluded that laboratory studies may overestimate the degree of alteration of psychomotor skills associated with exposure to trace concentrations of inhalation anesthetics.


Can the extent of halothane debromination be predicted preoperatively? In an attempt to predict the extent of halothane debromination, antipyrine metabolic clearance rate was measured preoperatively in 22 surgical patients, then correlated with percent hours of subsequent halothane exposure and postoperative serum bromide levels. There was a significant correlation (r = 0.78, p less than 0.001) between peak bromide level and anesthetic exposure but no correlation between peak bromide levels and antipyrine metabolic clearance rate. Thus, antipyrine is of no value for predicting the extent of halothane debromination. Mean peak bromide level 0.97 +/- 0.09 mM occurred on day 3 after anesthesia. This value is well below the psychoactive range. The data, therefore, also suggest that there is not a causal relationship between halothane biotransformation and the clinical syndrome of bromism.


Intravenous procaine as a supplement to general anesthesia for carbon dioxide laser resection for carbon dioxide laser resection of laryngeal papillomas in children. Procaine suppresses the cough reflex, decreases laryngeal irritability, and has general anesthetic properties. For these reasons, 14 pediatric patients undergoing CO2 laser resection of laryngeal papillomas were studied in which an intravenous infusion of procaine (1 mg/kg/min) was added to N2O-O2 halothane/enflurane general anesthesia immediately following endotracheal intubation. These patients were compared to nine patients receiving the same anesthesia without procaine. The mean age of both groups was 11 years. There was no difference between the groups in duration of anesthesia or surgery. Emergence, however, averaged 15 minutes in study patients compared to 36 minutes in the control group (p less than 0.01). There was no difference in anesthetic concentrations required to maintain satisfactory operative conditions in the two groups. Muscle relaxants were required intraoperatively in seven control patients but in none of the study patients. The surgeon ranked the operative conditions excellent in all study patients but poor in seven of the nine control patients. Five of the latter required postoperative treatment of laryngeal complications, including reintubation in three. Only one of the study patients had postoperative stridor. No evidence of procaine toxicity was noted in the study patients with total doses ranging from 500--3600 mg. Intravenous procaine is useful in pediatric patients having endoscopic laryngeal operations.


Antagonism of pancuronium and its metabolites by neostigmine in cats. Antagonism by neostigmine of neuromuscular blockade produced by pancuronium or its metabolites was studied in the cat anterior or tibialis muscle-peroneal nerve preparation using constant infusions of muscle relaxants. The ED50 of neostigmine (dose which caused a 50% antagonism) was 16, 11, 29, and 26 micrograms/kg for pancuronium, 3-hydroxypancuronium, 17-hydroxypancuronium, and 3, 17-hydroxypancuronium, respectively. Times of onset of neostigmine action were shorter when antagonizing 17-hydroxypancuronium neuromuscular blockade. Duration of neostigmine action when antagonizing 17- or 3, 17-hydroxypancuronium blockade was shorter than with pancuronium or 3-hydroxypancuronium. We conclude that more neostigmine is required to antagonize 17- or 3,17-hydroxypancuronium neuromuscular blockade than is required to antagonize pancuronium. Conversely, less neostigmine was required to antagonize 3-hydroxypancuronium blockade.


Pulmonary antibacterial defense mechanisms are depressed by halothane. Pulmonary antibacterial activity was determined by challenging halothane-exposed and control mice to radiolabeled (32P) Staphylococcus aureus produced by an aerosol generator and by determining the relative changes in viable bacterial and radioisotope counts in ground lung immediately after exposure and 4 hours later. Mice that were anesthetized for 4 hours with 1 or 2 MAC halothane and allowed to recover for 1 hour showed higher 4-hour bacterial counts when compared to controls, while radioisotope counts were the same in both groups. These data indicate that 1 and 2 MAC halothane anesthesia is associated with depressed lung bactericidal activity but that physical clearance mechanisms are not depressed.


Pancuronium reduces halothane requirement in man. We studied the contribution of pancuronium to anesthetic requirement in man by comparing MAC for halothane alone in 18 patients, with that after intravenous administration of pancuronium, 0.1 mg/kg, in 17 patients. In each group, movement was observed in limbs isolated from the circulation by tourniquet. In patients receiving pancuronium, halothane MAC was 0.55%, whereas the control value was 0.73%. Thus pancuronium reduces halothane MAC by 25%.


Incidence of fungal spores at the homes of allergic patients in an agricultural community. III. Associations with local crops. A predominantly agricultural community in California was surveyed for prevalent fungal spores during a 12-month period. Alternaria, Macrosporium and Stemphylium were recovered during asparagus and strawberry harvesting times year-round. Fusarium and Botrytis were less frequently associated with the strawberry harvest and were recovered only during the first quarter of the year. Epicoccum was recovered in the north end of the Salinas valley in low numbers throughout the year and was strongly associated with the strawberry and artichoke harvest. Aureobasidium (Pullularia) recovery occurred in different locations according to season, correlating somewhat with the cabbage harvest as well as with the harvest of strawberries. Recovery of the pigmented yeats showed strong correlation with the local growing season for lettuce. Cladosporium was prevalent year-round but did not appear to be signficantly affected by changing argicultural conditions. These data have permitted the predictability of mold aeroallergens with medical applications.


Impairment of tracheal ring ciliary activity by halothane. The effects of varying concentrations of halothane on tracheal ciliary activity were studied using a ferret tracheal organ culture technique. Using 1% and 2% halothane produced no impairment of ciliary activity, while 3% halothane caused some ciliostasis; 4% and 5% halothane produced rapid cessation of ciliary activity. Recovery was prompt and complete after exposure to 4% halothane for as long as 3 days, but after 4 days of exposure recovery was incomplete and after a week of exposure death of ciliary cells occurred.


Localisation of the NADPH delta 4-3-keto steroid-5 alpha-oxidoreductase in the rat ovary. The 5 alpha-reductase activity was localised on the stromal comportment of the rat ovary. The 5 alpha-reductase activity in stromal tissue decreased from 27.3 +/- 2.9 ng/hr/mg protein in rats 20 days old, to 13.4 +/- 1.2 ng/hr/mg protein in rats 30 days old and to 7.1 +/- 1.0 ng/hr/mg protein in adult rats. During the oestrous cycle no significant variations in enzyme activity could be demonstrated.


[Rapid change in the dynamics of antibody-forming cells under the influence of a single injection of various anthracyclines]. The dynamics of the titers and number of the antibody forming cells was studied within 26 hours after a single injection of anthracycline to immunized animals at various phases of the immune response. The specificity of the "rapid" effect of anthracyclines suggests that the low differentiated precursors of the antibody forming cells are the main target for the rubomycin effect and the immune competent cells in the state of multiplication and differentiation under the effect of the antigen are the main target for the carminomycin effect. The study was performed with noninbred mice immunized with sheep red cells.


Microbial transformations of natural antitumor agents: conversion of lapachol to dehydro-alpha-lapachone by Curvularia lunata. Microbial transformation of lapachol, a naturally occurring naphthoquinone, was carried out by Curvularia lunata (NRRL 2178). The fungus brings about oxidative cyclization of the substrate to dehydro-alpha-lapachone, which was isolated and characterized by nuclear magnetic resonance and mass spectral analyses; its structure was verified by chemical synthesis. The metabolite is a naturally occurring chromene possessing antibacterial and antitumor activities.


Nutritional status of light-for-date infants at birth and its influence on early postnatal growth. The ponderal index (PI) (g/cm3 X 100) was used to define the state of nutrition at birth of 47 light-for-date (LFD) term infants and to determine how intrauterine undernutrition influences growth during the first 6 months of postnatal life, at four age intervals: birth--4, 5--8, 9--12, 13--26 weeks. With the exception of one baby, each PI was less than 50th centile on the Miller and Hassanein standards: in 24 (51%) babies the PI was less than 3rd centile. This suggests that, generally, LFD infants are underweight for length, and by inference, are likely to have experienced intrauterine undernutrition. In the first month the 24 'wasted' infants (PI less than 3rd centile) gained more rapidly in weight, and grew more quickly in head circumference and length than the 16 'nonwasted' infants (PI greater than 10th centile). Thereafter growth rates were similar. With the exception of weight in the 'nonwasted' infants during the first month, rates of growth (weight, length, and head circumference) in both groups of babies in each of the 3 months after birth were greater than in normal infants. The first 3 months after birth can therefore be defined as the period of 'catch-up' growth in LFD term infants.


Unique amanitin resistance of RNA synthesis in isolated nuclei from Amanita species accumulating amanitins. In order to understand the basis upon which amanitin-accumulating species of Amanita are able to develop in the presence of these specific inhibitors of RNA synthesis, the in vitro RNA synthesizing activities of nuclei isolated from amanitin-accumulating species. Amanita hygroscopica (culture, derived from amanitin-accumulating carpophore) and A. suballiacea (carpophore), and from the non-accumulating species A. solitaria (culture) and A. brunnescens (carpophore) were tested for their sensitivities to alpha-amanitin inhibition. The nuclear RNA synthesizing activities obtained from both carpophores and cultures of non-accumulating organisms displayed significant sensitivities to alpha-amanitin, whereas those obtained from accumulating organisms displayed remarkable resistance to alpha-amanitin. The observed relationship between levels of amanitins in carpophores and resistance of the RNA synthesizing activities to inhibition by alpha-amanitin supports the hypothesis that amanitins may function as regulators of mRNA transcription in Amanita species.


[Ultrastructure of the adrenaline-synthesizing neurons of group A1 in the rat brain normally and under emotional stress]. Ultrastructure of A1 group, participating in the central regulation of cardio-vascular functions was studied in normal conditions and at early stages of an emotional stress. Normal adrenalin-synthesizing neurons of A1 group are characterized by a high intensity of processes of synthesis taking place in them. Abundant terminals, mainly adrenergic ones, which form varied contacts with neurons of A1 group, make a peculair feature. The hematoencephalic barrier in the area of A1 group as a sharply specific organization: transport of substances is mediated not only by pinocytosis, but also through fenestrated formations and through interendothelial spaces. Capillaries of the area studied have a powerful innervation--terminals immediately adjoin the vascular wall, sometimes penetrate into the basal membrane. An immediate contact between neurons and capillaries is noted. In the vicinity of the contact, the laminar complex is usually situated having increased in size vesicles, vacuoles, lysosomas, mitochondria; lobes of the invaginated nucleus are also directed towards this place. At an early stage of the emotional stress, when the arterial pressure is kept at the level close to the normal, rather essential changes are noted in neuronal ultrastructure, with significant spatial rearrangements including. Quantitative alterations of organells and reorganization of the neuronal ultrastructural apparatus results in formation of certain new properties and contribute to transformation of the neural cells to a more effective functional level.


Clinicopathologic correlation of xenon arc and argon laser photocoagulation. Procedure in human diabetic eyes. Xenon arc and argon laser lesions applied as scatter treatment to two diabetic patients were studied clinically and histopathologically. Seventy-one burns were grouped as very severe, severe, and moderately severe, according to pigmentation and diameter of the retinal scar. All lesions except half of the moderately severe ones showed full-thickness retinal involvement. In very severe xenon burns, whether xenon or argon, only proliferated retinal pigment epithelium provided a thin retinal scar, corresponding to an ophthalmoscopic appearance of "bare sclera". In the center of severe and moderately severe lesions, both glial cells and pigment epithelium bridged the retinal defect, forming a thick pigmented scar that contained pigment-laden macrophages. Rephotocoagulation over such thick scars may be less hazardous than is generally assumed. Nonvascular preretinal membranes were frequently present over very severe xenon lesions but rarely over very severe argon burns.


Non-surgical sterilisation of rams using a sclerosing agent. Injection of 0.25 ml of a solution of 3.6% formaldehyde in 90% ethanol direct into the caudae epididymides caused sterility in the rams treated. No live sperm were seen in ejaculates collected 35, 57, 91 and 196 days post-injection. The rams retained their libido and were mated to ewes between 2 and 3 months post-injection with no pregnancy ensuing. The technique offers a simple, safe, quick and effective method of sterilising rams. Attempts to produce sterilisation by injection of the sclerosing agent into the vas deferens were not successful.


[Diagnostic and therapeutic importance of breast pain]. In matters of mammary pathology, the attention of students has always been attracted by the study of tumours, while the varied field of mastalgia has been somewhat neglected. A series is reported in which a subdivision is suggested, on the basis of what has been done by other workers, whereby the various mammary pain syndromes are grouped. This is extremely useful, particularly for therapeutic purposes.


Effects of soy protein and saponins on serum, tissue and feces steroids in rat. Four groups of rats were fed, for 45 days, one of the following semipurified diets containing sucrose 55% (w/w) and (a) casein 25%, (b) casein 24%, saponins (from Saponaria officinalis) 1%, (c) isolated soy protein 25%, (d) soy protein 24%, saponins 1%. The soy protein diet, compared to the casein one, produced an increase in the fecal excretion of neutral sterols on the 29th and 42nd days, without any modification in the liver, aorta and serum cholesterol concentrations. The effect of soy protein cannot be attributed to its saponin content but other substances associated to soy protein may interfere. With the casein diet, added saponins increased the fecal excretion of neutral sterols and bile acids and decreased liver and aorta cholesterol levels. Serum cholesterol was found unchanged. The effects of saponins were suppressed or greatly reduced with the soy protein diet. These results could be explained by binding of the sterols in insoluble forms.


The location of Staphylococcus aureus in experimental chronic mastitis in the mouse and the effect on the action of sodium cloxacillin. Chronic staphylococcal mastitis was established in the mouse and electron miscroscopic examination of mammary tissue showed that the staphylococci were within epithelial cells, neutrophils and macrophages. Suspensions of cells containing staphylococci were prepared from chronically infected glands and incubated with sodium cloxacillin in vitro. This procedure resulted in a reduction in bacterial numbers, but a significantly greater reduction was obtained if the somatic cells were disrupted by sonication before the addition of cloxacillin. The results imply that the intracellular location of the staphylococci protects them against the action of the antibiotic.


In vivo effect of UV-B on lymphocyte function. Whole-body exposure of normal subjects to erythemogenic doses of UV-B radiation results in a decrease in the proportion of circulating E-rosette-forming lymphocytes, an increase in the proportion of null cells and a decreased incorporation of tritiated thymidine into DNA of lymphocytes following stimulation by phytohaemagglutinin (PHA). These alterations are dose-dependent, appear soon after exposure, reach a maximum after 8-12 h, and are reversed by 48-72 h post exposure.


Nitric oxide and carbon monoxide equilibria of horse myoglobin and (N-methylimidazole)protoheme. Evidence for steric interaction with the distal residues. The Soret absorption maxima and extinction coefficients of the CO and NO complexes of horse myoglobin and (NMeIm)protoheme (NMeIm = 1-methylimidazole) have been determined. The partition coefficient N, equal to the ratio P1/2 (CO)/P1/2(NO), has been determined spectrophotometrically for horse myoglobin and (NMeIm)protoheme. P1/2-(NO) values calculated from the partition coefficients are 5.7 x 10(7) mmHg for (NMeIm)protheme and 1.1 x 10(6) mmHg for horse myoglobin. The ratio of P1/2(NO) values for protein and model is 1.9 which is similar to a value of 1.6 reported for the ratio of P1/2(O2) values. These values may be compared to a ratio of 15 for CO binding to protein and model complexes. This different ratio for CO provides further evidence for steric interaction of the bound CO with the protein based on a consideration of the preferred nonlinear geometry of Fe-NO and Fe-O2 and the linear geometry of Fe-CO.


Conformational analysis by nuclear magnetic resonance: insulin. High-resolution 270-MHz proton nuclear magnetic resonance (NMR) spectra of the native two-zinc insulin hexamer at pH 9 have been obtained, and assignments of key resonances have been made. Spectra of zinc-free insulin titrated with Zn2+ are unchanged after the addition of 1 equiv of zinc per insulin hexamer, indicating that the conformation of the hexamer is fixed at this point and that the second zinc ion does not significantly change the conformation. Titration of the two-zinc insulin hexamer with anions high on the Hofmeister series such as SCN- causes marked changes in the NMR spectra which are interpreted as the result of major conformational changes to a new hexameric form of insulin having a twofold axis perpendicular to the threefold axis. Analysis of difference spectra indicates that this new hexamer (which should be capable of binding six zinc ions) binds 2 equiv of SCN- at two sites which are assumed to be identical and independent (K1 = 10(3), K2 = 2.5 X 10(2) M-1).


Protein-detergent interactions studied by capillary isotachophoresis. The interactions of sodium dodecyl sulphate with bovine serum albumin and ovalbumin have been studied by capillary isotachophoresis. This method makes it possible to determine very accurately the number of ligands bound to the high-affinity binding sites of the native protein. Bovine serum albumin was found to have seven high-affinity binding sites whereas ovalbumin in its native state was found to lack high-affinity binding sites for dodecyl sulphate.


Comparative studies of the action of long duration alcohol drinking on rat pancreatic exocrine cells. The influence of a long term ingestion of ethanol on the rat exocrine pancreas has been studied by electron microscopy. The comparison of micrographs from alcoholic and non alcoholic pancreas allowed us to compare the different organelles of acinar pancreatic cells. These data have been submitted to factorial analysis of correspondences. The ultrastructural modifications revealed out by this analysis are in good agreement with the hypothesis of an adaptation of exocrine cells to a chronic hyperfunctional state.


[Decrease in the stress reaction and the ulcerative lesions of the stomach under the influence of gamma-hydroxybutyric acid]. The effect of preliminary Na gamma-hydroxybutyric acid (GHBA) injection on the activation of adrenergic and hypophyseal-adrenal systems during emotional-painful stress and ulcerous affection of the stomach after the stress cessation was studied in experiments on rats. It was established that the preliminary GHBA administration inhibits stimulation of stress causing systems and thereby prevents ulcerous affection of the stomach mucosa. It is suggested that the activation of GHBA-ergic inhibiting system, which accompanies the stress, is a natural mechanism of limitation of the stress reaction and prevention of the stress affections.


[Effect of phenamine and caffeine on the behavioral changes occurring after the cessation of caudate nucleus electric stimulation in cats]. Low doses of amphetamine (0.1-0.5 mg/kg), not influencing the spontaneous behaviour of cats, prevent behavioral depression arising after repeated low-frequency stimulation of the caudate nucleus. Activating phenomena, observed after cessation of the stimulation, are considerably intensified up to the stereotypy formation. Caffeine (10-80 mg/kg), when activating spontaneous behaviour in cats, does not exclude a possibility of caudate-induced depression.


[Brain catecholamines at the early stage of emotional stress]. The blood pressure (BP) dynamics and catecholamine (CA) levels in the brain regions of August and Wistar rats were studied two hours after immobilization in order to elucidate the central neurochemical mechanisms leading to the destruction of BR self-regulation under emotional stress during the experiment. The BP level did not differ from the normal. The CA concentration in the hypothalamus, midbrain, isthmus rhombencephali and medulla oblongata underwent considerable changes. It is suggested that the mechanism of the CA changes in both strains of the rats exposed to stress is similar in the hypothalamus and medulla oblongata while both strains of the rats studied showed specificity in the activity of NA-synthesizing neurons of the isthmus rhombencephali and DA-synthesizing neurons of the midbrain that might be a cause of different resistance of cardiovascular functions during the later stage of immobilization.


Autonomic activity in subjects reporting changes in affect in the menstrual cycle. The paper reports a failure to find differences in autonomic activity in different phases of the menstrual cycle in subjects studied despite their reports of changes in affect.


Effect of intravesical formalin on the urothelium. In an experimental study, the urinary bladder of the rabbit was subjected to 10% formaldehyde instillation for periods ranging from 15 to 90 min. Varying degrees of cystitis, ranging from focal superficial to transmural necrotising cystitis, epithelial sloughs and intra-epithelial calcification may be seen. The depth of reaction was related approximately to the duration of exposure. Saline irrigation after instillation, contact time of 10 min and 1% formaldehyde seemed to reduce complications.


Basal bodies in the odontoblasts of the limpet, Patella coerulea L. (Gastropoda). The odontoblasts in the long radular gland of Patella coerulea L. are arranged in a terminal position; therefore newly formed teeth already have an upright position. The long and slender odontoblasts have only one to three lengthy and ramifying apical microvilli. Between these pinnate microvilli a fine filamentous material appears which probably corresponds to chitin microfibrils. Therefore, the pattern of chitin microfibrils seems to depend on the arrangement of odontoblasts' microvilli. For the first time, basal bodies were found in the apical part of odontoblasts which led to the assumption that the radular gland originally might have been a mucous gland, the secretion of which was transported by cilia.


Structural analysis of functionally different smooth muscles. The ultrastructure of the longitudinal and circular muscle cells of the guinea pig stomach which show different contractile responses was compared. The extracellular space within the muscle bundles is about 12.1% in the longitudinal layer and about 4.4% in the circular layer. Nexuses were sonsistently found in the circular muscle layer but not in the longitudinal muscle layer. Numbers of both mitochondria and microtubules per unit area of smooth muscle cell are larger in the longitudinal than in the circular muscle. Cellular area occupied by sarcoplasmic reticulum is about 4.7% in the longitudinal muscle, 2.3% in the circular muscle. The numbers of caveolae are almost the same in both tissues. The most distinct difference between the two types of smooth muscle is the appearance of the thick filaments. The circular muscle cell contains approximately 50 thick filaments per 0.5 micron 2 of cytoplasmic area, while the longitudinal muscle cell only about 25 filaments which were usually much thinner than those of the circular muscle. These results indicate that the contractile apparatus itself is different in longitudinal and circular smooth muscles of the guinea pig stomach.


The comparative ultrastructure of the egg membrane and associated pore structures in the starry flounder, Platichthys stellatus (Pallas), and pink salmon, Oncorhynchus gorbuscha (Walbaum). Eggs of the starry flounder (Platichthys stellatus) and pink salmon (Oncorhynchus gorbuscha) were examined by scanning and transmission electron microscopy to determine differences in egg membrane structure with reference to contrasting ecological conditions in which the eggs normally develop. The egg membrane of the starry flounder constitutes 0.22--0.50% of the egg's diameter. The zona radiata is composed of 6 continuous horizontal lamellae, covered by a thin triple layered border, and pierced by numerous regularly spaced pore canals. The micropyle canal measures 8 microns at the opening and tapers to 3.6 microns as it penetrates the membrane. In contrast, the thicker membrane of the pink salmon egg forms 0.80--1.0% of the egg's diameter, is composed of numerous short discontinuous lamellae which are pierced by pore canals, and is covered by a coating of irregular thickness. The 15--16 micron micropyle opening is surrounded by an area of protrusions, and the funnel-shaped canal tapers to 2 microns at its terminal aperture. Contrasting environmental conditions during embryogenesis of these two species may be reflected by the thin membrane and simple lamellar structure in the pelagic egg of the starry flounder, and the thick membrane and complex lamellar structure in the demersal egg of the pink salmon.


The lamina propria of the bovine seminiferous tubule. The boundary tissue of bovine testicular seminiferous tubules exhibits remarkable regional differences at the level of the seminiferous tubule proper, as compared with its terminal segment. The basal lamina of the seminiferous tubule proper is multilayered and possesses knob-like protrusions. At the level of the terminal segment the basal lamina is highly specialized; in the region of the terminal plug candelabrum-like projections of the tubular basal lamina invade the bases of the modified supporting cells up to a depth of 3.5 microns. The adjoining surface of these supporting cells is densely studded with hemidesmosomes. The elongated peritubular cells are arranged in 3--5 concentric layers around the tubulus seminiferus proper but form a loose association at the level of the terminal segment. Where the terminal segment joins the testicular straight tubule, peritubular cells may assemble to constitute a contractile spiral. Elastic tissue is situated mainly subjacent to the tubular basal lamina and to a lesser degree between the peritubular cell layers. A peritubular space lined by endothelium-like cells may surround the seminiferous tubule proper and also the transitional zone of the terminal segment.


Ultrastructure of the heart of the sturgeon Acipenser stellatus (Chondrostei). The ultrastructure of atrial and ventricular myocardial cells from Acipenser stellatus is described. The cells of the atrium are more loosely connected than those of the ventricle. Cell contact is by simple intercalated discs and by desmosomes. The cells are flattened, with peripheral myofibrils and a central region of mitochondria and the nucleus. The sarcoplasmic reticulum consists of subsarcolemmal tubules, that frequently extend towards the central mitochondria. Dyads are small and positioned at any sarcomeric level. No T-tubules are present. Specific granules are restricted to the atrial cell, and are sometimes present within the SR tubules.


The fine structure of the cumulus oophorus during follicular development in sheep. The cumulus and membrana granulosa of non-atretic ovarian follicles from primordial up to a stage shortly before ovulation were studied by electron microscopy. The follicular cells of primordial follicles were undifferentiated and rested on a thick basal lamina. In secondary follicles the endoplasmic reticulum had proliferated forming an anastomosing network. In early antral and antral follicles (0.5--2.0 mm dia.) the ER was composed of short cisternae, the mitochondria had elongated and gap junctions were first observed. In late antral follicles (3.0--5.9 mm dia.) gap junctions were frequent. In the cumulus the glycogen was associated with electron lucent areas whereas in the granulosa it was invariably associated with membranes. In large antral follicles large membrane bound bodies were present in the basal cells of the cumulus. At early oestrus a distinctive mitochondrial morphology was noted in the granulosa but not elsewhere in the follicles. At mid oestrus numerous annular nexuses were present in the granulosa but not in the cumulus. At late oestrus numerous lipid droplets were formed in both cumulus and granulosa, the boundary with theca interna became indistinct and the basal lamina became incomplete.


Nuclear inclusions in paraventricular nucleus neurons of the rat hypothalamus. This paper deals with the ultrastructure of two types of intranuclear inclusions, "microfilamentous spindle-shaped" and "crystalloid", present in paraventricular nucleus neurons of adult normal rats. These inclusions appear occasionally in some non-secretory neurons of the parvocellular system, but have never been seen in neurosecretory cells of the magnocellular system. The microfilamentous spindle-shaped inclusions show a close spatial relationship with the granulofibrillar body and interchromatin granules. The distribution and functional significance of such structures are discussed in the light of recent ultrastructural and biochemical studies on nuclear inclusions.


Junctions between interstitial cells of the renal medulla: a freeze-fracture study. Cell junctions between interstitial cells of the renal medulla were studied in freeze-fracture replicas of kidneys from rat, rabbit, hamster and the tree-shrew Tupaia belangeri. In all species studied a composite type of intercellular junction was found comprising elements of tight junctions and irregular gap junctions of highly variable size and shape. The number of these junctions increased towards the tip of the papilla. Our findings suggest that the composite junctions observed play a role in the maintainance of the ladder-like arrangement of the interstitial cells in the inner zone. The existence of irregular gap junctions raises the possibility that the functions of the interstitial cells are coordinated, especially during alterations of the functional state of the kidney.


An actin filament matrix in hand-isolated nuclei of X. laevis oocytes. The nuclear gel of Xenopus oocytes contains a meshwork of randomly oriented microfilaments which have been identified as F-actin by decoration with rabbit skeletal muscle myosin subfragment-1 (S-1). Nuclear gel preparations treated with S-1 differ in several respects from control preparations incubated in either aqueous medium alone, or medium containing BSA. Actin filaments in control preparations appear less well preserved than those in S-1 treated preparations of the nuclear gel. The nucleoli of control preparations are extremely dense, while those of S-1 treated preparations have a more open, granular appearance. Large granular aggregates, which are a prominent feature of the controls, are seen much less frequently in S-1-treated preparations of the nuclear gel. These morphological differences appear to be correlated with the binding of protein to F-actin, since nuclear gel preparations incubated in tropomyosin, which also binds to actin filaments, appear similar to those treated with S-1. Approximately 63% of the total nuclear actin exists in a globular state, while 37% is filamentous.


Oocyte-follicle cell relationships in a starfish. The follicle cells which surround the oocytes of starfish are known to both release the hormone 1-methyladenine and to respond to it by an active movement which forms a component of the spawning response to the hormone. In Patiria miniata these flagellated cells are located peripheral to the oocyte and have long cytoplasmic processes which penetrate the vitelline layer to the egg surface to form an adhering zonule-like junction. Within the follicle cell cytoplasm are located elongate filamentous bands which appear to represent a component of the contractile mechanism that mediates follicle cell response to 1-methyladenine. These bands do not resemble the filaments of vertebrate smooth muscle cells (quantity, distribution and size of filaments; lack of dense bodies in the filament mass), nor the contractile units of the superficial epithelium of lower vertebrate follicles.


Evidence for a DNA ligase change related to early cleavage in axolotl egg. A definite change in the forms of DNA ligase appears when the axolotl egg enters cleavage. Sucrose gradient and phosphocellulose chromatography show that the a 6S form of DNA ligase exists before division, i.e. in unfertilised and fertilised egg, and a 8.2S form is present at the first division. N-ethylmaleimide sensitivity and heat stability are different for the two forms. The possible significance of this early change is discussed.


Idiopathic oedema: a study of the effects of bromocriptine. We have studied the effect of bromocriptine in seven patients with idiopathic oedema. Two patients were unable to tolerate the drug; the remaining five were maintained on doses of between 5 and 7.5 mg a day. Symptomatic improvement was noted in two patients and in both of these and one other there was a significant improvement in mean diurnal weight gain. The fall in free water and sodium clearance on standing was reduced by bromocriptine, but there was no significant change in the excessive fall in plasma volume.


Heterozygote detection in glucose-6-phosphate dehydrogenase deficiency: limitation of hair follicle analysis. Glucose-6-phosphate dehydrogenase deficiency was demonstrated in a case of favism. The X-linked enzyme defect was expressed in erythrocytes but not in hair root cells. Predictably, the mother shown to be a heterozygous carrier on the basis of intermediate erythrocyte glucose-6-phosphate dehydrogenase activity could not be identified as a carrier by means of hair root study. It seems to be necessary to test the hair roots of at least one enzyme-deficient member of the family to exclude false negative results, if hair root analysis is used for carrier detection. Because of the more or less clonal origin of hair roots, they remain a convenient biopsy material with which to study heterozygosity in X-linked inborn errors of metabolism.


Diurnal variation in uterine estrogen receptors in immature female rats--inhibition by arginine vasotocin. A diurnal variation in uterine estrogen receptors from immature female rats was observed with the peak occurring during the mid-light phase (noon) of the cycle. Although no dose response was noted, all concentrations of arginine vasotocin ranging from 5 x 10(-7) to 5 x 10(-9) significantly inhibited binding of 3H-estradiol to estrogen receptors in the cytosol fraction of uteri obtained from immature female rats either during the light or dark phase of the photoperiod.


Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl)-maleimide. Glucose oxidase from Aspergillus niger was conjugated with rabbit immunoglobulin G or its monovalent fragments (Fab'). The enzyme was treated with N-hydroxysuccinimide ester of N-(4-carboxycyclohexylmethyl)-maleimide to introduce maleimide groups, which were then allowed to react with thiol groups of reduced IgG or Fab'. More than 40% of immunoglobulin G, Fab' and enzyme used could be conjugated without self-coupling. The enzyme activity decreased about 26 and 15% upon conjugation with immunoglobulin G and Fab', respectively, and the ability of antibody to bind to antigen was well preserved in conjugates. Conjugate preparations purified by gel filtration contained little free form of immunoglobulin G, Fab' or enzyme. Both the cross-link and enzyme activity in Fab' conjugate were stable at pH 6-7 at 4 degrees C for at least 6 months.


Conidiogenous loss of structuro-functional polarity in the hyphal tips of Sclerotinia fructigena. Ultrastructural depolarization in the conidiogenously induced hyphal tips is initiated by Spitzenkörper (apical body) disintegration, followed by spreading of wall-associated vesicles and migration of mitochondria and lipid granules into the exclusion zone of the vegetative apices.


Dietary cyanide and tropical malnutrition diabetes. Two categories of diabetes are recognized in the temperate zone--ketosis-prone diabetes requiring insulin and diabetes not requiring insulin. Another unique type of diabetes occurs in the tropics. It has two forms, both different from either form of temperate zone diabetes. Type J and pancreatic diabetes are both characterized by youth onset, antecedent malnutrition, substantial insulin requirement, and resistance to ketosis. In the tropical countries where they are found, both forms are associated with specific dietary practices, including a nutritionally marginal protein intake. The close association with low protein intake distinguishes this form of diabetes from that occurring in North America, Europe, and Oceania. The geographic distribution of malnutrition diabetes, in addition to being limited to the tropics, coincides regularly with the consumption of tapioca (cassava) or other foods that contain cyanide-yielding substances. Ingested cyanide is normally detoxified, principally, by conversion to thiocyanate. This detoxification requires sulfur, derived principally from amino acid sources. Studies in the rat indicate a remarkable ability to detoxify ingested cyanide, a reduction in urinary thiocyanate excretion when protein intake is lowered (especially during growth), production of marked hyperglycemia by either oral or parenteral cyanide, and the development of cyanosis and epidermal changes when there is prolonged exposure to cyanide. Both the association of malnutrition diabetes with food cyanogens and our laboratory observations support a role for cyanide in its pathogenesis.


Fine structure of the mitotic cycle of unfertilized sea urchin eggs activated by ammoniacal sea water. Unfertilized sea urchin eggs enter a mitotic chromosome cycle after treatment with sea water containing ammonia. Centrioles cannot be found but microtubules are formed in the later stages of the cycle. The microtubules are displayed in an astral arrangement centered on clusters of osmiophilic bodies. In early stages, distinct kinetochores on the condensed chromosomes show no attachments to microtubules. Later, a few microtubules may be attached to the kinetochores. The chromosomes and microtubules are contained in a "clear zone", a large compact accumulation of membranes which displaces yolk particles and mitochondria, but not ribosomes, from that region of the cell. No bipolar spindle is formed.


Filaments associated with the endoplasmic reticulum in the streaming cytoplasm of Chara corallina. Perfused Chara cells capable of resuming ATP-dependent cytoplasmic streaming in low free Ca++ solutions have been examined by electron microscopy for myosin-like filaments. Filaments 44 nm in diameter and up to 3 micron in length have been found associated with the endoplasmic reticulum that along with mitochondria, microbodies and dictyosomes from the endoplasm becomes immobilised around the sub-cortical actin bundles when ATP is depleted. Such endoplasmic filaments have not been detected in association with mitochondria or microbodies and they have not been found in the stationary cortex. These filaments are extracted from the perfused cell by ATP unless motility-inhibiting levels of cytochalasin B are present. The filaments are not detectable in cells inactivated in solutions containing high (10(-4) M) Ca++ concentrations even when the Ca++ level is subsequently lowered. Consistent with their being required for motility, cytoplasmic streaming cannot be effeiciently reactivated by ATP in such filament-depleted cells. The possibility is discussed that the filaments contain myosin and that the endoplasmic reticulum with which they are associated has a major role in generating and transmitting the motive force for streaming.


The effects of tannic acid on the in vivo preservation of microfilaments. A population of single, actin-like microfilaments has been found specifically associated with the cortical microtubules or radish root hairs. Extensive filament bundles are not specifically associated with the tubules although close proximity is observed. A technique is described for obtaining a reliable quantitative estimate of the size of these microfilament populations. The addition of tannic acid to the fixative preserved more microtubule-associated microfilaments, without revealing any extra non-microtubule-associated filaments. Microfilaments preserved with tannic acid have diameters which increase in proportion to the concentration of tannic acid used.


Long-term response to human growth hormone in 36 children with idiopathic growth hormone deficiency. The results of long term treatment with human growth hormone (Crescormon, 12.4 IU/m2/week) in 36 patients with idiopathic growth hormone deficiency are given. Birth trauma--in particular assisted breech delivery (30%)--is the major aetiological cause. Twelve patients had isolated growth hormone deficiency (IGHD), 24 had multiple pituitary hormone deficiencies (MPHD) of which 19 were treated with additional thyroid hormones. The results were judged by the criteria of height velocity, total height gain and change of height prediction (TW2, age based). It is concluded that the growth hormone dose chosen in many cases is insufficient to maintain high growth rates after the first year of treatment, when "catch-up" no loner takes place. The tendency of patients supplemented with thyroid hormone to grow better--without additional bone-age advancement--calls for careful search for hypothyroidism and suggests the use of thyroxin in cases of doubt.


Age differences and stress ulcer in the rat. Rats 2-, 7- and 12-mo. old were exposed to supine body restraint plus cold (5 degrees C) for 3 hr. The mean cumulative length of lesions for the 2-, 7-, and 12 mo. old rats was 22.0, 43.0 and 16.0 mm. respectively. The same experimental design was used in a second study, but the pylorus was ligated prior to restraint. Total acid output/hr. was 134.6, 178.2, and 64.7 muEq/60 min. respectively for the three age groups. Older rats were not more susceptible to stress-ulcer and gastric acid was not significantly related to degree of ulceration.


Presence of spermatozoa in cervicovaginal smears from young and old women. Routine cervicovaginal (PAP) smears from 1,239 women from ages 16 to 92 were examined retrospectively for spermatozoa. The prevalence of spermatozoa in cervicovaginal smears was found to be highest (25%) in the 20-24 year age group, and fell gradually to 20% in the 70-79 year age group. The oldest woman in the study to have spermatozoa in her smear was 76 years old. In order to evaluate persistence of spermatozoa in the lower genital tracts of women, 697 smears were prepared from women who had volunteered the time of last coitus, birth control methods, and douching practices. Spermatozoa could be found in smears taken up to 10 days after coitus, although their number decreased due to vaginal douching and birth control. These findings confirm that coital activity in aging women can persist into the eighth decade of some women's lives.


Pattern of follicular growth and steroidogenesis in the ovary of aging cycling rats. Prior to the cessation of reproductive cycles, older female rats exhibit irregular and prolonged cycles due to alterations in the hypothalamic-pituitary-ovarian axis. In order to evaluate the age-related changes in the ovary, the histology, and estradiol, testosterone and progesterone concentrations within the ovaries of mature regular cycling (4--5 mo. old) and older irregular cycling (10--11 mo. old) rats were examined. At estrus, the number of non-atretic growing follicles (150--300u in diameter) was greater in the mature than in the older rats (18 +/- 1.5 vs 4.5 +/- 1.4). However, the number of preovulatory follicles on proestrus did not differ (6.0 +/- 1.2 vs 5.5 +/- 0.6). Estradiol, testosterone and progesterone concentrations on proestrus in mature rats averaged 38.8 pg, 56.1 pg, and 1.0 ng/ml of ovary, respectively. In the older proestrous rat, only estradiol was altered, increasing to 124.3 pg/mg. In addition, many of the preovulatory follicles within the aged ovary were larger (greater than 600u in diameter) than those within the mature ovary. On the day of estrus virtually all preovulatory follicles ovulated in the mature rat, whereas large follicles, less than or equal to 600u in diameter, remained in the older ovary. In addition, estradiol levels remained elevated and ovarian cysts were observed in the aged ovary. Thus, in the older irregular cycling rat, 1) pre-ovulatory follicles develop, but many do not ovulate; 2) these non-ovulatory follicles form ovarian cysts which remain within the ovary. The number of cysts may increase with age until a polycystic ovary develops and the rat enters a constant estrous state.


Ontogeny of behavioral development, arousal and stereotypes in two strains of mice. A number of reflexes and amphetamine-induced locomotor and stereotyped behavior were assesses in 8, 16, 32, 90 and 360 day old C57BL/6J and SEC/1ReJ inbred mice. The data indicate that C57 mice are more precocious for a number of neuronal and behavioral mechanisms while SEC mice are less mature at birth. In addition, there are appreciable fluctuations of these behavioral patterns throughout life. A rise in arousal levels was evident in both strains between 8 and 16 days and between 32 and 90 days of age. Three-hundred-sixty days old mice presented a general decrease in the levels of arousal. These findings are discussed in terms of neuronal and behavioral plasticity and in relation to the ontogeny of the different catecholaminergic systems which modulate excitory and inhibitory different behavioral patterns at different ages.


A prodrug of ADTN: selectivity of dopaminergic action and brain levels of ADTN. The effects of administration of the prodrug dibenzoyl ADTN (DBADTN) on ADTN concentrations in rat brain and on behaviour in rats having a unilateral 6-hydroxydopamine lesion in the corpus striatum have been studied. Using a combination of HPLC and electrochemical detection as assay method it was found that there was a more selective accumulation of ADTN in the corpus striatum than in the cerebellum. In addition the accumulation of ADTN in the corpus striatum was slow in onset yet long in duration. The peak concentration of ADTN was relatively low and although it was sufficient to cause a strong stimulation of presynaptic DA receptors it did not cause significant rotation in the unilaterally lesioned rat. Prodrug methodology may thus prove useful in designing new selectively acting DA agonists.


Effect of bromocriptine on gonadal ornithine decarboxylase in vitro. The alterations in ornithine decarboxylase activity in response to in vitro treatment with bromocriptine suggest the possible direct action of the drug on gonads.


Diurnal rhythm of active calcium transport in rat intestine. The active calcium transport along the small intestine of adult rat was shown to exhibit daily rhythmic variations of considerable amplitude. Both the light-dark cycle and the time of food presentation acted as synchronizers of the rhythm observed.


[Diurnal variations evidence of glomerular filtration in the rat (author's transl)]. Parallel variations in plasma and urine variations of endogenous urea and creatinine level in the rat during 4 consecutive 6-h-long periods permit to evidence urea and creatinine clearance diurnal variations with very significant increase in the 2 nightly periods. The signification of this very large nightly glomerular filtration increase is discussed.


[A radioimmunoassay for plasma parathyroid hormone (PTH) using N-terminal PTH antiserum (author's transl)]. In order to investigate plasma bioactive PTH, we tried to assay the N-terminal portion of PTH by RIA. The antiserum to PTH was prepared by immunizing rabbits with a bovine 1-34 PTH conjugate BSA. A preparation of labeled PTH was radioiodinated by the chloramine-T or lactoperoxidase method. Labeled PTH was purified by means of adsorption by Quso G-32 powder or a sephadex G-50. The separation of the free and bound labeled hormone was performed by the dextran-coated charcoal method. The assay was carried out as follows: 0.2 ml diluted buffer (0.05 M, pH 8.6, veronal buffer), 0.1 ml standard PTH or sample to be tested, and 0.1 ml anti-PTH serum were mixed. After the first incubation at 4 degrees C for 4 days, 0.1 ml labeled PTH were added. After a second incubation at 4 degrees C for 12 hours, the assay tubes were centrifuged at 2,000 rpm for 30 min and the precipitates were counted. Various hypothalamic, pituitary and thyroid hormones did not interfere with the RIA for PTH. A dose response curve was obtained in a range from 100 pg to 5,000 pg per ml of standard PTH in this assay system. The serum immunoreactive PTH in healthy subjects values less than 290 pg per ml.


Maternal-channelled infant feeding.

Maternal-channelled infant feeding. Proper feeding of the mother during pregnancy and lactation -- preferably with locally available foods -- will optimize the volume and composition of breast milk (both as the sole food for the young baby and as a supplement for the transitional). This approach will avoid the economic, infective and distributive complexities of introducing cow's milk and bottle feeding unnecessarily, and will also help to retard the decline in breast feeding on a community basis.


Neonatal behavior of infants of adolescent mothers. The Brazelton Neonatal Assessment Scale was administered to 30 infants of adolescent mothers (under 18 years of age at delivery) and 30 infants of older mothers, at between two and five days of age. The infants of the adolescent mothers were found to be significantly less capable of responding to social stimuli, less alert, and less able to control motor behavior and to perform integrated motor activities. The results are further discussed in terms of the 'stimulus value' of the infant and areas of focus for efforts at early intervention.


Epidemiology of myelomeningocele in Utah. This study suggests that in the Intermountain area of the United States (Utah, Wyoming, Idaho and Montana) there is an effect of social class on the incidence of myelomeningocele: the higher the social class the less the incidence. This survey also suggests that there is an influence from the social class in which the mother was raised, as well as that into which she marries. Birth order and maternal age were found to have no influence on the incidence of myelomeningocele in this study.


[Formation of structural chromosome mutations in metaphase of mitosis]. The rate of structural chromosome mutations at metaphase of the first mitosis was determined in culture of embrionic mouse fibroblasts after UV-irradiation during the S-period (lambda = 265 nm at an incident dose of 40 erg/mm2). It is established that the mutation rate is higher at late metaphase than at early metaphase. After the cell treatment with intercalating compounds (actinomycin D, acridine orange or ethidium bromide) at metaphase, the rate of UV-induced chromosome aberrations was decreased (about 2-fold). It is concluded from the results obtained that the majority of aberrations arise during metaphase after UV-irradiation in the process of DNA synthesis. After the cell treatment with o-methylhydroxylamine (OMHA) during the S-period the rate of structural mutations was the same at late and early metaphases. This rate was not affected by the caffeine treatment at metaphase; during this stage the acentric chromosome fragments lie outside the equatorial plate, which is an indication that the OMHA-induced aberrations, in contrast to the UV-induced aberrations, are formed before the beginning of metaphase, possibly during the interphase. It is suggested that the chromosome condensation during metaphase is of importance in the formation of structural mutations.


Night-day variations of renin-activity in primary aldosteronism. To detect changes in previously unmeasurable low renin activity plasma specimens of 20 patients with primary aldosteronism (12 with an unilateral adenoma and 8 with idiopathic bilateral adrenal hyperplasia), obtained at short term intervals between 20.00 and 8.00, were incubated over a prolonged period of 18 hours. 6 of 12 patients with an aldosterone producing adenoma (APA) and 3 of 8 patients with idiopathic bilateral adrenal hyperplasia (IAH) showed typical night-day variations of PRA with lower values before and higher values after midnight. 7 of these 9 patients with night-day rhythmicity of PRA simultaneously showed secretory episodes. In 2 patients (1 with APA, 1 with IAH) PRA was constantly undetectable (less than 0.2 ng/ml . 18 h) and in 2 patients with APA a fixed secretion of renin was observed. We failed to demonstrate typical night-day variations of PRA in 3 patients with APA and in 4 with IAH, although in 5 of these 7 patients secretory episodes of PRA were found. Our results show that different patterns of PRA curves may be observed both in patients with APA and IAH. Thus, analysis of PRA curves is of no value to differentiate patients with APA from those with IAH.


Radioimmunoassay of unconjugated and total serum estetrol using a 125I-iodinated tracer. A radioimmunossay (RIA) for the measurement of both unconjugated and total serum estetrol has been developed, using an antiserum to an E4-3-conjugate and a 125I-radioiodinated E4 tracer. Assay of dried ethyl ether extracts was used for the determination of unconjugated E4, while a direct measurement of unextracted hydrolyzed serum in the presence of 0.3%  8-anilino-1-naphthalene sulphonic acid (ANS) proved adequate for total E4. Assay reliability was evaluated and the procedure standardized through a series of tests aimed at assessing accuracy, sensitivity and precision. No steroidal interference was found to practically affect the assay (0.3% estriol cross-reactivity), nor were solvent and sample blanks observed in the case of unconjugated E4. For total E4 assay, the sample blank effects were acceptably overcome by using hydrolyzed male serum and 0.3% ANS, as a standard diluent. An interassay variability amounting to approximately 10 and 6% resulted for unconjugated E4 and total E4 RIA, respectively. A number of serum samples (285 for unconjugated E4, 147 for total E4) randomly collected throughout normal pregnancy were assayed. The unconjugated E4 levels at 15th week and at term were 62.7 +/- 22.6 and 766.5 +/- 208.2 (SD) pg/ml, respectively. Total E4 was about 6--7 times higher than the levels of free E4 and increased 7 times from the 15th week to term.


Interaction of steroid-binding serum proteins with concanavalin A-Sepharose 4B. The interaction between steroid-finding serum proteins and concanavalin A was studied using affinity chromatography on concanavalin A-Sepharose 4B. The sex hormone-binding globulins of man and rabbit together with the corticosteroid-binding globulins of man, rabbit and guinea pig were all bound quantitatively by the affinity medium. Only two thirds of rat corticosteriod-binding globulin showed an affinity for concanavalin A indicating this protein to be heterogeneous with respect to terminal mannose or glucose residues. The progesterone-binding globulin in pregnant guinea pig serum, possessing the highest carbohydrate content of the proteins investigated, did not bind to concanavalin A-Sepharose 4B. Affinity chromatography on concanavalin A-Sepharose 4B is documented to be a valuable supplement to existing methods for analysing steroid-protein interactions.


The immune response of pigs to infection with the stomach worm Hyostrongylus rubidus (Hassall and Stiles, 1892). III. Effect of a primary infection on immunoglobulin-positive cell numbers. An immunofluorescent study has been made of IgG, IgM and IgA in the stomach of uninfected minimal disease pigs and pigs infected with a single dose of 100,000 third stage infective Hyostrongylus rubidus larvae. Immunoglobulin-positive cells were present in both uninfected and infected animals. In infected animals both IgM and IgA immunocytes increased in numbers following infection and the numbers remained raised during the histotropic phase of worm development. No increase in IgG immunocyte numbers was noticed throughout the course of infection (100 days).


[Postoperative serum immunoglobulin G, prealbumin, retinol-binding protein and transferrin picture]. In 24 patients who had undergone cholecystectomy or abdominal hysterectomy the serum levels of prealbumin, transferrin, retinol-binding protein and immunoglobulin G were determined up to the 5th day after operation. Intra- and immediately postoperatively the patients received only isotonic infusion, beginning on the 3rd day after operation, they were fed increasingly orally. The observed serum levels decreased postoperatively.


[Possibilities of calculating the fat-free body mass and its reaction to a carbohydrate-poor, fat-rich diet]. 9 healthy medical students got an isocaloric fat-rich carbohydrate-reduced diet during 14 days. Body-weight and lean body mass were registrated. We calculated lean body mass from measurement of skinfold thickness, total body potassium and total body water. Body-weight was reduced highly significantly by 3.4 kg on an average. Body potassium and body water remained unchanged. It can be concluded from these results that weight loss was caused by an exclusive reduction of the fat mass.


[Tolerance studies of an intravenous fat emulsion (FE-S15) with beagle dogs]. The potential toxicity of FE-S15, a soybean oil fat emulsion used in parenteral nutrition, was studied in dogs. Forty pure bred beagles, divided into two experimental groups (FE-S15 at 9 and 4 gm/kg BW/day) and two corresponding control groups (receiving Dextrose Ringer's Solution) were given daily infusions for 28 days via a central venous catheter. When compared with control groups no significant weight loss was observed in either experimental group; the food intake decreased only in animals receiving fat in high doses. Hemoglobin and hematocrit decreased in all groups, the greatest fall observed in the group receiving high dose fat infusion was the hematocrit decline from 43.9% to 31%. This decrease was significantly different from the control only during one observation period. The total serum lipids, triglyceride and phospholipid concentrations of the animals receiving fat in high doses increased 3-4 times in comparison to that of the control group; cholesterol increased 5 times. The serum protein level fell from 6.5 to 5.1 gm/dl in animals receiving 9 gm/kg BW/day while animals receiving 4 gm/kg BW/day had a significant increase to 8.4 gm/dl. Except for an overall decreased activity clinical sign such as lethargy, loss of appetite, vomiting, and diarrhea were infrequent and equally observed in experimental and control animals. The post mortem examination did not reveal changes that must be attributed to the administered fat. It is concluded that the fat emulsion FE-S15 is fairly well tolerated in dogs at a potentially toxic level.


Effect of succinylation on the oligomeric structure of glycinin. By varying the ratio of succinic anhydride to the protein, glycinin, one of the major fractions of soybean proteins, is succinylated to various levels. Sedimentation velocity experiments indicate the dissociation of the protein due to succinylation. Viscosity increases and a blue shift occurs in the absorption spectrum. The rate of proteolysis increases. Both dissociation and denaturation of the protein appear to occur. The effect of syccinylation on glycinin and arachin, the major protein of groundnuts, appears to be different.


Application of the aluminum-formaldehyde (ALFA) histofluorescence method for demonstration of peripheral stores of catecholamines and indolamines in freeze-dried paraffin-embedded tissue, cryostat sections and whole-mounts. This paper describes new procedures for highly sensitive visualization of monoamine stores in peripheral tissues, taking advantage of the recently introduced aluminum-catalysed formaldehyde (ALFA) reaction. The tissues are exposed to an aluminum sulphate solution (with or without formaldehyde fixation) in a perfusion and/or immersion step, followed by formaldehyde vapour treatment. Procedures are described for freeze-dried, paraffin embedded tissue, cryostat sections and whole mount preparations. For all these tissue preparations the ALFA method gives a highly sensitive and precise demonstration of catecholamine-containing neurons and 5-HT-containing cells in a variety of peripheral tissues. For freeze-dried tissue and cryostat sections the ALFA method represents an improvement in comparison with other available methods. This is particularly noticeable for the very delicate adrenergic nerves in such organs as the thyroid, ovary, pancreas and the gastrointestinal tract.


Cobalamins and cobalamin-dependent enzymes in Candida utilis. Candida utilis has been shown to contain 4.7 pmol of cobalamin per g of wet cell paste. Purification of the cobalamin showed it to be a mixture of methylcobalamin and adenosylcobalamin. Two cobalamin-dependent enzyme systems have been found in the yeast: methylcobalamin-dependent methionine biosynthesis and leucine 2,3-aminomutase. The cobalamin extracted from the yeast is as effective as authentic adenosylcobalamin in stimulating leucine 2,3-aminomutase.


The effect of side chain structure of ester substrates in determining the rate-controlling step in alpha-chymotrypsin-catalyzed hydrolysis. Presteady state and steady state analyses of the alpha-chymotrypsin [EC 3.4.21.1]-catalyzed hydrolysis of three specific ester substrates and three ring-substituted derivatives were carried out to elucidate the effect of hydrophobic interactions due to the different side chains of the substrates on the individual steps of the reaction. Hydrolysis of all the substrates except for N alpha-acetyl-Nin-formyltryptophan methyl ester (Ac-Trp(CHO)-OMe) was controlled by the deacylation rate. In spite of their comparable Ks values, the substrates with small kcat, such as N alpha-acetyltryptophan methyl ester and N alpha-acetyl-2-(2-nitro-4-carboxyphenylsufenyl)-tryptophan methyl ester, characteristically gave Km values one order of magnitude smaller than the others. For the reaction of Ac-Trp(CHO)-OMe, it was ascertained that the deacylation step was not rate-controlling. It is suggested that the acylation step controls the rate in this case.


Mechanism of association of a specific aldehyde inhibitor, leupeptin, with bovine trypsin. Leupeptin (acyl peptidyl-L-argininal) is a potent inhibitor of trypsin and related proteases. We analyzed the association of leupeptim with bovine trypsin kinetically, assuming that it proceeds by a pathway which involves two steps: E + I in equilibrium K1 Complex I k-2 in equilibrium k+2 Complex II. The observed dissociation constant (K1) for the first step was 1.24 X 10(-3) M (at pH 8.2 15 degrees C) and the two first-order rate constants (k+2 and k-2) were 166 s-1 and 1.75 X 10(-3.s-1, respectively (at pH 8.2, 15 degrees C). The dissociation constant (Kd) for the whole process was calculated from these parameters to be 1.34 X 10(-8) M. This value is compatible with that determined directly by an independent static method (2.36 X 10(-8) M). We also measured Kd for the leupeptine complex of anhydrotrypsin, a trypsin derivative in which the active-site hydroxyl group is missing. The observed value was about 5 orders of magnitude larger than Kd and was rather similar to K1 in native trypsin. A elupeptin isomer which contains a D-argininal residue did not show strong affinity towards trypsin. These findings suggest that complex II consists of a covalent hemiacetal adduct formed between the serine hydroxyl group in the enzyme active site and the aldehyde group in the inhibitor. The pH dependencies of the dissociation constant and other parameters show that deprotonation of the charge-relay sustem in the active site is important for the formation and stabilization of complex II.


Purification and characterization of a lectin from the mushroom, Flammulina veltipes. A lectin was purified to homogeneity from the mushroom, Flammulina veltipes, by zinc acetate treatment and CM-cellulose column chromatography. Its molecular weight was estimated to be 20,000 by gel filtration and polyacrylamide gel electrophoresis. The lectin does not contain carbohydrate, half-cystine, methionine, or histidine. On gel filtration sith Sepharose 6B in the presence of 6M guanidine-HCl, the purified lectin dissociated into two nonidentical subunits, FVA-L (molecular weight, 12,000) and FVA-S (8,000). The hemagglutinating activity was retained only in the FVA-L subunit. The lectin is mitogenic with respect to mouse spleen lymphocytes.


Mouse gamete interactions during fertilization in vitro. Chlortetracycline as a fluorescent probe for the mouse sperm acrosome reaction. We have developed an assay for detecting the acrosome reaction in mouse sperm using chlortetracycline (CTC) as a fluorescent probe. Sperm known to be intact with nonreacted acrosomes show CTC fluorescence in the presence of Ca2+ over the anterior portion of the sperm head on the plasma membrane covering the acrosome. Sperm which have undergone the acrosome reaction do not show fluorescence on the sperm head. Mouse sperm bind to zonae pellucidae of cumulus-free eggs in vitro in a Ca2+-dependent reaction; these sperm are intact by the CTC assay. Intact sperm bind to mechanically isolated zonae under the same conditions: the egg is apparently unnecessary for this inital reaction. Sperm suspensions, in which greater than 50% of the motile population had completed the acrosome reaction, were prepared by incubation in hyperosmolal medium followed by treatment with the divalent cation ionophore, A23187. Cumulus-free eggs challenged with such sperm suspensions preferentially bind intact sperm; acrosome-reacted sperm do not bind. We conclude that the plasma membrane of the mouse sperm is responsible for recognition of the egg's zona pellucida and that the obligatory sequence of reactions leading to fusion of mouse gametes is binding of the intact sperm to the zona pellucida, followed by the acrosome reaction at the zona surface, followed in turn by sperm penetration of the zona.


Evidence for the participation of actin microfilaments and bristle coats in the internalization of gap junction membrane. Thin sections of rabbit granulosa, human SW-13 adrenal cortical adenocarcinoma, and mouse B-16 melanoma cells revealed an apparent single-layered basket of 4- to 7-nm filaments surrounding cytoplasmic gap junction vesicles. This interpretation was based upon apparent longitudinal, cross, and en face sections of structures surrounding the vesicle profiles in tissue treated with tannic acid-glutaraldehyde. In granulosa cells incubated with the S-1 fragment of heavy meromyosin, arrowhead-decorated filaments were observed at the periphery of the gap junction vesicles, suggesting that these filaments contained actin. In addition, we found that small gap junctional blebs and vesicles at the cell surface were coated with short electron-dense bristles similar in appearance to the cloathrin-containing coat of coated vesicles of nonjunctional membrane. The role of these and other cytoskeletal elements in the possible endocytosis of gap junction membrane is discussed.


Species-specific sperm adhesion in sea urchins. A quantitative investigation of bindin-mediated egg agglutination. Bindin, a protein component of the acrosomal vesicle of sea urchin sperm, has been isolated from Arbacia punctulata and strongylocentrous purpuratus. Using this isolated bindin, we have devised a quantitative assay for bindin-mediated egg agglutination and compared the agglutination of bindin eggs from A. puntulata and S. purpuratus. Bindin- mediated agglutination is species -specific in both species, although a measurable degree of heterotypic interaction is observed. Homotypic bindin-egg interactions differ significantly from heterotypic interactions both in the extent of agglutination and the size of the resulting aggregates. We also provide direct evidence that bindin particles agglutinate eggs by adhering to the surfaces of adjacent eggs. Although the A. punctulata bindin preparation displays the same functional properties and consists of one major polypeptide of the same apparent molecular weight as S. purpuratus bindin, its morphology is very different. Unlike the spherical aggregates observed with S. purpuratus bindin, A punctulata bindin exists as lamellar vesicles and binds significant amounts of phospholipids and Triton X-100, suggesting that it may be tightly associated with the acrosomal membrane. Having defined a number of the basic parameters of bindin-mediated agglutination, we examined the effect of a number of saccharides and glycopeptides on bindin-mediated egg agglutination. Carbohydrate-containing components derived from the egg cell surface by proteolysis were found to inhibit bindin-mediated egg agglutination at low concentrations, but this inhibition is not species specific.


The growth and differentiation of transitional epithelium in vitro. The development of rat transitional epithelial cells grown on conventional non-permeable surfaces was compared with development on permeable collagen supports. On glass or plastic surfaces, cells grew as expanding nomolayer sheets. Once confluent, growth continued with a bilayer being formed in most areas and apical cells being continuously sloughed off. Although most cells were interconnected by desmosomes, and junctional complexes were formed, no other indications of differentiation were observed. After 2-3 wk of growth, division stopped and cel death ensued. In contrast, single-cell suspensions plated on collagen-coated nylon disks reassociated into multicellular islands and commenced growth. Mitoses were confined to the basal cells in contact with the permeable substrate. The islands developed into epithelial trilayers, tapering to monolayers along spreading edges. Once the islands were confluent, stratification was completed and appeared similar to that observed in vivo. Germinal cells formed a basal lamina, and the upper layer was composed of large, flattened cells with an unusually thick asymmetrical plasma membrane on the apical surface. Electron microscopic and radioactive tracers demonstrated "leaky" zonulae occludentes with a restricted permeability to small molecules. The movement of urea was retarded in comparison to water. Unlike the slow turnover of adult epithelium in vivo, maturation and sloughing of apical cells were measurable. Transfer of cells could be effected and growth maintained for up to 4 mo. These results may indicate the necessity of a nutrient-permeable growth surface for the polarized differentiation of adult transitional epithelium.


Cytochalasin inhibits the rate of elongation of actin filament fragments. Submicromolar concentrations of cytochalasin inhibit the rate of assembly of highly purified dictyostelium discoideum actin, using a cytochalasin concentration range in which the final extent of assembly is minimally affected. Cytochalasin D is a more effective inhibitor than cytochalasin B, which is in keeping with the effects that have been reported on cell motility and with binding to a class of high-affinity binding sites from human erythrocyte membranes (Lin and Lin. 1978. J. Biol. CHem. 253:1415; Lin and Lin. 1979. Proc. Natl. Acad. Sci. U.S.A. 76:2345); 5x10(-7) M cytochalasin B lowers it to 70 percent of the control value, whereas 10(-7) M cytochalasin B lowers the rate to 25 percent. Fragments of F-actin were used to increase the rate of assembly fivefold by providing more filament ends on to which monomers could add. Under these conditions, cytochalasin has an even more dramatic effect on the assembly rate; the concentrations of cytochalasin B and cytochalasin D required for half-maximal inhibition are 2x10(-7) M and 10(-8) M, respectively. The assembly rate is most sensitive to cytochalasin when actin assembly is carried out in the absence of ATP (with 3 mM ADP present to stabilize the actin). In this case, the concentrations of cytochalasin B and cytochalasin D required for half-maximal inhibition are 4x10(-8) M and 1x10(-9) M, respectively. A scatchard plot has been obtained using [(3)H]cytochalasin B binding to F-actin in the absence of ATP. The K(d) from this plot (approximately 4x10(-8) M) agrees well with the concentration of cytochalasin B required for half-maximal inhibition of the rate of assembly under these conditions. The number of cytochalasin binding sites is roughly one per F-actin filament, suggesting that cytochalasin has a specific action on actin filament ends.


Identification and organization of the components in the isolated microvillus cytoskeleton. We have examined the effects of ATP and deoxycholate (DOC) on the cytoskeletal organization of Triton-demembranated microvilli (MV) isolated from chicken intestine brush borders. Isolated MV are composed of a core of tightly bundled microfilaments from which arms project laterally to the plasma membrane with a 33-nm periodicity. These lateral arms spiral around the core microfilaments as a helix with a 25 degrees pitch. Demembranated MV consist of four polypeptides with mol wt of 110,000, 95,000, 68,000, and 42,000, present in molar ratios of 1.1:1.6:1.3:10.0. After addition of 50 microM ATP and 0.1 mM Mg++, the cytoskeletons are organized as a tight bundle of microfilaments from which lateral arms are missing. In these ATP-treated cytoskeletons, the 110-kdalton polypeptide is reduced in amount and the 95,000, 68,000, and 42,000 polypeptides are present in a 1.3:1.2:10.0 ratio. In contrast, after incubation with 0.5% DOC, the core microfilaments are no longer tightly bundled yet the lateral arms remain attached with a distinct 33-nm periodicity. These DOC-treated cytoskeletons are depleted of the 95,000 and 68,000 polypeptides and are composed of the 110,000 and 42,000 polypeptides in a 2:10 molar ratio. These results suggest that the microfilaments are associated into a core bundle by the 95- and 68-kdalton polypeptides and from this core bundle project the lateral arms composed of the 110-kdalton polypeptide.


Mitochondria and mitochondria-tonofilament-desmosomal associations in the mammary gland secretory epithelium of lactating cows. The lactating cow mammary secretory epithelial cell is very active synthetically and contains numerous very pleomorphic mitochondria. Cup- and ring-shaped mitochondria are frequent and many are extremely elongated. Preferential localization of mitochondria in the basal region, or at the lateral margins of the secretory cell adjacent to intraepithelial monocytes have been observed. Occasional mitochondria show one or several very densely staining cristae with a lattice pattern. Mitochondria are frequently seen closely associated with the tonofilament-desmosomal complex. These structures and associations are present after immersion or perfusion fixation of mammary gland from several breeds of cow; their possible significance is discussed.


The effect of colchicine on fibrillar material in wheat meiocytes. Wheat meiocytes contain intranuclear bundles of microfibres at stages from pre-meiotic mitosis until zygotene. This includes early pre-meiotic interphase, the stage when colchicine induces univalents at subsequent metaphase I. This paper reports the effect of colchicine on the structure and distribution of such fibrillar material. Many normal-looking bundles of intranuclear fibrillar material formed after prolonged colchicine treatment (up to 5 days) which totally suppressed microtubule assembly in meiocytes. Thus, fibrillar material is unlikely to comprise tubulin. However, the formation of intranuclear fibrillar material was retarded in colchicine-treated meiocytes, and fibrillar material was absent during the colchicine-sensitive stage for univalency. The occurrence of cytoplasmic fibrillar material was delayed and greatly increased in colchicine-treated meiocytes compared with controls. A novel curved form of fibrillar material was seen in colchicine-treated meiocytes. It is concluded that colchicine had significant effects on the assembly and distribution of fibrillar material. These results are, therefore, compatible with the possibility that colchicine causes univalency in wheat via its effects on fibrillar material.


Ethyl acetate as a substitute for diethyl ether in the formalin-ether sedimentation technique. Ethyl acetate appears to be a satisfactory subsitute solvent for diethyl ether in the Formalin-ether sedimentation technique. In comparative studies, concentration of organisms with ethyl acetate was equal to or greater than that with diethyl ether. No distortion or alteration of morphology was observed with eigher solvent, and preparations were comparable in appearance and ease of examination. In addition, ethyl acetate is less flammable and less hazardous to use than diethyl ether.


The role of hyperdipsia in aggression following septal lesions in rats. Lesions of the septum in animal subjects are known to produce an increase in aggressiveness and an increase in water intake. A series of experiments was carried out to examine the possibility that aggression was secondary to hyperdipsia. When rats with septal lesions were restricted to preoperative levels of water intake, aggression scores declined significantly. When animals without lesions were preloaded, with either water or saline, aggression increased. Neither decreased shock threshold nor increased cell hydration provided a full explanation for the results. It is suggested that the aversive nature of the stomach turgescence caused by increased water intake may be an additional mediating factor in septal aggression.


Help-agents' views about clinical interactions with acting-out children. A scale to measure nonprofessional child-aides' views about, and practices in, working primarily with acting-out children is described. The scale was used as part of an overall evaluation of a program to teach the aides Ginottian limit-setting approaches for work with such children. A prior study had shown that the training was followed by significantly more favorable treatment outcomes. Compared to 44 nontrained child-aides, the 19 trained aides had significantly higher postprogram scale scores on opinions and beliefs about acting-out children, changes in feeling about working with them, and actual observed changes in their playroom behaviors. Item analysis indicated that, following training, aides felt more comfortable with and had a richer repertoire of techniques for dealing with acting-out children. Specifically, they found it easier to set limits and to deal with overtly aggressive behaviors. Those changes may be key factors in explaining the significantly greater effectiveness of the trained aides working with acting-out children.


[Treatment of the epithelial cysts of the iris by Argon laser photocoagulation (author's transl)]. In this work seven cases with epithelial cyst of the iris resulting from trauma or cataract surgery were studied and treated by Argon laser photocoagulation. The authors analyse theirs results stressing the fact that no alteration of the lens or the cornea has been provoked after a follow-up of three years.


Freeze substituted tissue in 5'-nucleotidase histochemistry. Comparative histochemical and biochemical investigations. The suitability of freeze-substitution in n-butanol and paraffin embedding of tissues for the histochemical demonstration of 5'-nucleotidase was investigated and compared with commonly used preparation techniques, such as fresh frozen sections and cryostate sections of cold formalin and glutaraldehyde-fixed rat liver. The influences of each step of the preparation techniques on the enzyme activity were controlled by a quantitative radiochemical assay. Freeze substitution was revealed to be superior to the commonly used preparation techniques with respect to: 1) high sensitivity and specificity of the histochemical 5'-nucleotidase reaction (this is based on the fact that incubation media with very low lead concentrations (0,6 mM/1) can be used); 2) excellent morphological appearance of the tissues showing cytological details of enzyme localization; 3) unlimited storage of the tissue materials and ease of sectioning.


Attempt to isolate antierythrocyte antibodies by the method of affinity chromatography. II. IgG - incomplete antibodies. It ensues from the results of the present work that erythrocyte stromata can serve as a suitable insoluble immunosorbent of both complete and incomplete antibodies. When applying optimum conditions, such as laboratory incubation temperature and suitable buffers, it is possible, by means of a chromatographic column with 0.5 ml erythrocyte stromata suspended in 40 g of glass beads (ballotini), to obtain fractions devoid of incomplete antibodies. Incomplete antibodies could be absorbed completely, either directly by a 24-hour incubation of the respective serum on the column or by passaging the serum through the column. In the presence of 0.2 M NaCl, absorption of incomplete antibodies was prevented; only complete antibodies were demonstrated in the fractions of the eluates. The elution yield in incomplete antibodies was low; in some cases, it could not be demonstrated at all. It can be explained partly by non-specific absorption, partly by the low initial antibody titre. It is therefore rather difficult to separate the individual incomplete antibodies from each other in this method. The column with incorporated erythrocyte stromata can be recommended for incomplete antibodies only because of its absorption capacity (to remove incomplete antibodies), but not for obtained elution yield.