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.

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.


Prevalence of Coxiella burnetii infection among humans and domestic animals of Rajasthan State, India.

A total number of 1806 sera comprising 1049 humans and 757 animals from four ecologically different areas of Rajasthan State were tested to determine the prevalence of complement-fixing (CF) antibodies to Coxiella burnetii (C burnett). Of the 1049 human and 757 animal sera tested, antibodies to C. burnetii were detected in 195 sera (18.6 per cent) and 187 sera (24.7 per cent), respectively. Among humans, the prevalence of infection with C. burnetii was highest in the desert area of Barmer district and the hilly area of Bundi district, viz 64.5 per cent and 28.2 per cent, respectively. It was slightly lower in the semi-arid and the arid zones of the State, viz Sirohi (20 per cent) and Jalore (13.7 per cent), whereas the irrigated areas of Kota and Jaipur had the lowest prevalence, 9.9 and 8.4 per cent, respectively. Among animals, the highest prevalence was detected in sheep (39.6 per cent), followed by cattle (29.9 per cent) and goats (18.6 per cent).


Basic criteria and classification scheme for bacteriophages acting against cholera vibrios of the classical and El Tor biotypes.

In accordance with the principles, nomenclature and taxonomic tests recommended by the TCNV for the systematization of viruses, cholera and El Tor phages were classified in all categories of the taxonomic hierarchy. On the basis of this classification, all known cholera bacteriophages could be affiliated to the biological type Vira, sub-type Deoxyvira, class 2/D B -- Deoxybinala, family Phagoviridae, genus Phagovirus vibrio and species Phagovirus cholerae. Phagovirus cholerae includes homologous series III-D, IV-D and V-D with 10 serologic types representing the smallest taxonomic units. At same time, phages of the remaining families with a single-strainded structure of DNA and RNA have so far not been detected for cholera vibrios.


Litter sizes in inbred strains of rats (Rattus norvegicus).

The average litter sizes and reproductive performance of twenty-five inbred strains of rats, maintained in a conventional colony in Pittsburgh, and twenty-six inbred strains, maintained in a barrier facility at the National Institutes of Health, were calculated from data collected for periods of 3 and 10 years, respectively. There were considerable variations in both parameters in each of the two colonies studied. In addition, there were quite wide variations in male to female ratios among inbred strains (0.80 to 1.42).


Serum levels of gonadotrophins in hamsters exposed to short photoperiods: effects of adrenalectomy and ovariectomy.

Female hamsters rendered acyclic by exposure to short photoperiods (10 h light: 14 h darkness) showed diurnal surges of both LH and FSH which persisted after removal of the ovaries and adrenal glands. The reduced increase in gonadotrophin secretion after ovariectomy, typical of hamsters exposed to short days, was also not contingent upon the presence of the adrenal glands. The results demonstrate that 'supersensitivity' of the neuroendocrine axis to feedback of ovarian or adrenal steroids cannot fully account for either the daily surges of gonadotrophins or the diminished response of LH and FSH to ovariectomy in hamsters maintained on short days. It is suggested that these changes result instead from other pineal-mediated actions of short photoperiods upon the brain.


Effects of aromatizable androgens on aggressive behaviour among rats (rattus norvegicus).

Three experiments were used to tests the applicability of the aromatization hypothesis of androgen action to aggressive behaviour among Norway rats. In Expt 1, administration of testosterone propionate was highly effective in restoring aggressive behaviour to castrated rats while 17 beta-hydroxy-5 alpha-androstan-3-one was of intermediate effectiveness. Of the steriods tested in Expt 2, androstenedione and testosterone were highly effective, 17 beta, 19-dihydroxyandrost-4-en-3-one was of intermediate effectiveness and cholesterol was ineffective. The results of Expt 3 indicated that treatment with testosterone or oestradiol both resulted in increased aggression while treatment with (5 alpha,17 beta)-17,19-bis(acetyloxy)-andostan-3-one diacetate (5 alpha-19-hydroxytestosterone) was without effect. Androgens which were aromatizable and could be 5 alpha reduced, i.e. testosterone, testosterone propionate and androstenedione, were highly effective in restoring aggressive behaviour; however, two other steroids, 5 alpha,19-hydroxytestosterone which is 5 alpha reduced, and 19-hydroxytestosterone, which can be aromatized, were respectively of low or medium effectiveness on behaviour. However, oestradiol, which did not maintain sexual development of accessory glands, was highly effective in the restoration of aggressive behaviour. Since the behaviourally active steroids in the present experiments were not only those predicted by the aromatization hypothesis, it is proposed that several steroids are capable of activating aggressive behaviour and that the aromatization hypothesis does not adequately explain the hormonal basis of aggressive behaviour among Norway rats.


Progesterone receptors and RNA polymerase activity in the rat uterus during the oestrous cycle.

The concentrations of progesterone receptors in uterine cytoplasm and nuclei were measured during the oestrous cycle of the rat. The concentration of cytoplasmic progesterone receptors was highest at pro-oestrus and declined at oestrus to reach lowest levels at metoestrus before rising at dioestrus. Similar changes were observed in the concentration of nuclear progesterone receptors, the highest levels being present at pro-oestrus and dioestrus. In addition, both activities A and B of RNA polymerase mirrored these alterations in nuclear receptor levels.


A radioimmunoassay for neurotensin in human plasma.

A specific radioimmunoassay for neurotensin in human plasma has been developed capable of detecting changes of 5 pmol/l with 95% confidence. Neurotensin-like immunoreactivity has been detected in human plasma in two molecular forms and rises by 27 +/- 8 (S.E.M.) pmol/l (n = 9) after ingestion of food.


Effects of the method of exsanguination on measurements of developmental changes in serum concentrations of follicle-stimulating hormone and luteinizing hormone in prepubertal and adult male house mice.

The relationship between the method of exsanguination and the serum concentration of gonadotrophins was investigated in male house mice. Mice were killed 21-99 days post partum by ether anaesthesia and cardiac puncture, or by decapitation. Results of gonadotrophin radioimmunoassays indicated that titres of serum FSH were not affected by the method of exsanguination, while concentrations of serum LH were significantly raised when blood was collected by ether anaesthesia and cardiac puncture. Serum FSH in an inbred strain of mice peaked at 21 days post partum and declined steadily to a relatively constant pubertal and postpubertal level. Concentrations of serum LH were variable during the prepubertal period with a peak occurring 31--35 days post partum. Extremely high titres of FSH on days 21-35 post partum indicated that mice in this study might be relatively insensitive to FSH in the prepubertal period.


Examination of the role of 'non-functional' corpora lutea in the female rat.

In rats with 5 day reproductive cycles, 'anovulatory' cycles were induced by blockade of ovulation with sodium pentobarbitone injected at pro-oestrus. Such anovulatory cycles were characterized in the ovaries by gradual atresia of the large follicles present at the time of the injection and the growth of a new cohort destined for the next ovulation. In the vaginal smears, anovulatory cycles were indistinguishable from normal ovulatory cycles. The serum concentrations of progesterone remained at baseline levels during dioestrus of anovulatory cycles whereas increased concentrations of progesterone were observed during dioestrus of ovulatory cycles. It is concluded that the 'non-functional' corpora lutea of the cycle are the source of dioestrous progesterone. The length of anovulatory cycles after a single injection of pentobarbitone was 5 days despite the absence of any increase in progesterone concentrations during dioestrus. It is concluded that progesterone production during dioestrus plays no major role in the control of the duration of 5 day reproductive cycles.


DNA synthesis after polyspermic fertilization in the axolotl.

Cytological and autoradiographic studies were done to investigate the cytoplasmic control of DNA synthesis under conditions of physiological polyspermy. The DNA synthetic phases of the egg, principal sperm and accessory sperm nucleic were determined and correlated with nuclear morphology and develpmental fate. Results show that accessory sperm nuclei undergo morphological transition to pronuclei. Their DNA synthetic phase is the same as that of the principal sperm nucleus. Hence accessory sperm nuclei are capable of initiating and completing DNA replication before any cytological evidence of their degeneration is observed.


Naloxone blockade of apomorphine-induced stereotyped behavior: interaction of endogenous opiates with dopamine.

The specific opiate antagonist, naloxone, inhibits the in vivo and in vitro activity of the endogenous opiate compounds which have heretofore been identified. In this study systemic naloxone administration successfully blocked the production of stereotyped behavior induced by the direct dopamine agonist apomorphine. This implies that the endogenous opiates contribute to the production of stereotyped behavior initiated by dopaminergic stimulation and that endogenous opiates may function as central neurotransmitters with dopaminergic activity.


Studies on soybean factors which produced goiter in rats.

The goitrogenicities of various preparations obtained from soybean were studied using rats. Thyroid weight, 131I uptake and the ratio of 131I-monoiodotyrosine to 131I-diiodotyrosine of the gland in rats fed the preparation was measured to determine the activity of these preparations. It was resulted that the group of rats receiving the curd had the most hypertrophied thyroid, the enlargement being equal to that in the group of these receiving soybean in the 8 weeks' feeding experiment. No difference on the content of iodine were found with the various diet preparations. The proteolytic digestion of soybean curd did not eliminate the goitrogenic property, thus, the goitrogen seems not to be a protein or peptide-like substance.


Structure--activity relationships of n-substituted dopamine and 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene analogues: behavioral effects in lesioned and reserpinized mice.

N,N-Disubstituted dopamine and 2-amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (6,7-ADTN) analogues were synthesized and tested intraperitoneally in mice for dopamine agonism. Compounds inducing asymmetric postures in unilaterally caudectomized mice were further tested in mice treated with reserpine and alpha-methyl-p-tyrosine methyl ester. ED50 values determined for reversal of reserpine-induced catalepsy were used to rank drug potency and correlated with molecular structure. N-n-Propyl N-substituted compounds were more effective than other N,N-dialkyl homologues. Of these, analogues with one alkyl group larger than propyl became inactive or their dopaminomimetic effect was reduced when the propyl was replaced with a larger group. N-Monosubstituted analogues were inactive as dopamine agonists. N-n-P-r-N-n-Bu-6,7-ADTN was six times more potent than N-n-propyl-N-n-pentyl- and N-n-propyl-N-phenethyldopamine but ten times less potent than apomorphine. The availability of an array of structurally related dopamine analogues with dopaminomimetic properties may make it possible to test the hypothesis that there are more than one type of dopamine receptor and that stereotypy and locomotor activity may have different central nervous system loci. Moreover, they may be of potential use in the treatment of parkinsonism and other extrapyramidal disorders.


An objective method for measuring fluorescence of individual sperm cells labelled with 1-anilinonaphthalene-8-sulphonate (ANS) by means of photomicrography and densitometry.

A method is presented for measuring the degree of fluorescence of sperm cells labelled with 1-anilinonaphthalene-8-sulphonate (ANS). Labelled spermatozoa were subjected to incident-light u.v. illumination and the fluorescent image was photographed. Densitometry of the photographic film yielded values for individual sperm heads, and the mean value for ten cells formed the unit of data for parametric statistical analysis. The method is both objective and precise, and was used to distinguish between treatments applied to the spermatozoa. It is believed that the method has a wide application as an inexpensive means of fluorimetry.


Staphylococcus aureus mastitis in nursing mink affected with aleutian disease.

An outbreak of staphylococcal mastitis in nursing female ranch mink (Mustela vison) is described. Lesions were acute necrotizing mastitis, fatty infiltration of the liver and renal tubules, and adrenal cortical hyperplasia. The presence of Aleutian disease in the herd suggests a role of immunosuppression in the outbreak.


Epidemiologic aspects of american Kaposi's sarcoma.

In our series of 37 patients with Kaposi's sarcoma, a larger than expected incidence of Americans of Italian and Jewish lineage was found, with 51% of Italian and 38% of Jewish background. Although a predominance of these two groups has been clearly shown previously, this may be the highest percentage of any series. In addition, 12, or 32%, of the patients were found to have concurrent diabetes mellitus. This association has been observed previously, but is not generally appreciated.


Effect of acute and chronic ethanol intake on metastasis formation in rat liver.

The effect of acute ethanol intoxication and prolonged intake of ethanol on metastasis formation in the liver was studied in the rat. The tumour cells used were derived from a pharmacologically induced colon carcinoma (NGW 1) and were injected intraportally. Acute ethanol intoxication at the time of injection was associated with a liver weight of 13.29 +/- 0.89 gm two weeks later, compared to 8.43 +/- 0.51 gm in controls. Daily intake of ethanol for eight weeks resulted in a mild reduction of platelets. The liver weight in two groups was 7.31 +/- 0.50 gm and 12.19 +/- 1.40 gm three weeks after intraportal injection of cancer cells as compared to 9.53 +/- 1.11 gm and 19.63 +/- 1.67 gm in controls. The results indirectly supported the hypothesis that acute ethanol intoxication causes increased platelet activity and confirmed earlier results showing the importance of platelets for metastasis formation.


Prolactin: its role in the development of mammary tumours.

Experimentally, it has been found that the rate of DNA synthesis of normal mammary glands which is a limiting factor for mammary tumourigenesis is largely dependent upon the circulating level of prolactin. Based on these results, one of the roles of prolactin in the development of mammary tumours is hypothesized to create mammary gland conditions favourable for the action of carcinogens through its stimulation of the rate of mammary gland DNA synthesis, a measure of the frequency of mammary gland cell division.


[Pathogenesis of urinary stress incontinence and principles of its surgical treatment].

As in different types of stress incontinence the mechanism of the destroyed urethral support is not the same, the principles of their surgical treatment are bound to differ. Generally speaking, the surgical intervention in stress incontinence aims at securing adequate mobility of the lower urinary organs, forming an effective suburethral support, and correcting the elements of the prolapse. Applying these principles in 508 women with stress incontinence having been surgically treated at the University Hospital Department of Gynecology and Obstetrics in Skopje from 1968 to 1976 and followed up at least two years afterwards, the global rate of relapses was 6.1%, by which the best results were obtained by Burch's modified method (2.1%), while after operations after Bonney or Barnett the frequency of relapses was relatively higher (7.1%). Analysing technical details in the carrying out of the above mentioned procedures, the authors plead for a most adequate preoperative preparation of patients, so as to determine and categorize the type of stress incontinence in question and allow the choice of the most appropriate surgical intervention in any given case.


The effect of exposure fractionation on the yield of nondisjunctional segregation involving compound X and free 4-chromosomes in Drosophila.

This paper describes the effect of fractionation of an exposure of 2000-R X-rays into two 1000-R exposures separated by a 3-h interval on the frequency of various segregation products following chromatoid interchange between a compound X and the 4th chromosome. It was observed that fractionation leads to a decrease of all exceptional progeny, the result being significant at the 0.05-probability level for both classes of detachments; i.e., with or without a 4th chromosome. The finding that the same effect of fractionation was observed for numerical aberration (non-disjunction) and detachment supports the idea that these reflect different consequences of the same primary effect, radiation-induced rearrangement. Furthermore, alternative recoveries were observed to occur at equal frequencies. Following C(1)-4 interchange, the univalent 4 does not segregate at random, but assorts apart from its homologue 1.5 as frequently as it moves to the same pole.


Effect of nitrites and nitrates on irrigation and secretion of gastric mucosa in rat.

An aspect of the vasomotor effect of nitrites and nitrates was studied by investigating the influence of these substances on the mucosal blood flow and hence on gastric secretion in the rat. Firstly, point measurements were obtained using an aminopyrine clearance technique after intravenous nitrite and nitrate treatment. Then with an appreciably different technique (14C-aniline clearance on perfused stomach) the kinetics of the phenomenon were followed after, firstly, intravenous administration of the two agents and secondly their intragastric administration. It is interesting to note that nitrites and nitrates are active to differing degrees and present opposing effects: nitrites decrease whereas nitrates globally enhance the factors under consideration (gastric mucosal blood flow, gastric juice volume, acory effect and cuase no modification on intragastric administration. The flow rate-gastric parameter correlations are significant in all cases and show modifications of the aqueous compartments of the body.


Natural changes in the cell division rate in normal tissues and the possible role of adrenalin concentration.

Mitotic incidence (MI) and DNA labeling index (LI) were measured in the liver, lung and kidney of rats at different times throughout the day. There were significant variations in MI and LI in all organs and there were also significant differences between organs in respect of the pattern of variation. It was found that adrenalin had no consistent inhibitory action on the MI and LI of these tissues maintained in organ culture. These results are not consistent with the view that diurnal (or circadian) rhythms in mitotic activity are directly mediated by changes in adrenalin secretion. In organ culture the rate of entry into DNA synthesis was higher than the rate of mitosis, leading to significant increases in the proportions of cells with reduplicated DNA content.


Evidence that chronic apomorphine treatment enhances 5-hydroxytryptamine metabolism in brains of normal and neonatally hyperthyroid rats.

Daily injection of L-triiodothyronine (10 microgram/100 g, s.c.) for 30 days to neonatal rats significantly enhanced the metabolism of 5-hydroxytryptamine as reflected by increased tryptophan hydroxylase activity and 5-hydroxyindoleacetic acid levels of certain discrete brain regions. However, neonatal L-triiodothyronine treatment produced no change in 3H-5-hydroxytryptamine uptake by crude synaptosomes. Chronic treatment with apomorphine (1 mg/kg/day, s.c.) for 15 days, beginning from 15 days of age, increased tryptophan hydroxylase activity as well as 5-hydroxytryptamine and 5-hydroxyindoleacetic acid levels and blocked the uptake of 3H-labelled serotonin in crude synaptosomes of normal and L-triiodothyronine-treated animals. Furthermore, apomorphine (which is known to indirectly stimulate 5-hydroxytryptaminergic neurons) produced a greater increase in tryptophan hydroxylase and 5-hydroxyindoleacetic acid in the mid-brain region of neonatally hyperthyroid animals as compared to normal rats. These data indicate that excess thyroid hormone in early life not only increases the turnover of brain 5-hydroxytruptamine, but also enhances the sensitivity of dopamine receptor sites. thus amplifying the stimulating action of apomorphine. Our findings also suggest that thyroid hormone in early life advances the overall development of monoaminergic systems in the brain.


[The epicutaneous DNCB test and CEA determination in neoplasms of the breast, digestive tract, lung and urogenital system].

110 patients observed over a three-year period suffering from cancer of the breast, digestive apparatus, lung and urogenital apparatus, and submitted simultaneouly to the DNCB skin test and to radioimmunological measurement of CEA have been studied. Confirming what has already been reported, the epicutaneous findings and laboratory results show there is an almost constant correlation between marked reactivity to DNCB and normal CEA titre and that, on the contrary, ml response to immune stimulation is often matched by a pathological CEA value. It is maintained that this type of cross investigation may be important for prognosis as the basis for initial assessment of the probable clinical course of the disease and the response of the patient to immunochemotherapeutic treatment.


Evaluation of indications for cataract surgery.

The large number of cataract operations performed in the United States annually has raised questions as to the indications and methods of documentation. The criteria designed for the Physicians and Surgeons Review Organization (PSRO), the thrust of second opinions, and an evaluation of the types of cataracts producing visual impairment are discussed. The contralight test of Junker is demonstrated as a means of determining the effect of cataracts on visual acuity and could be used as an indicator of the need for surgery.