[Effects of biotechnical management procedures on blood parameters in gilts. 2. Protein-bound iodine and copper]. Two groups of twelve gilts each kept on platforms were synchronised in two passes over 20 days, using 100 mg Suisynchron per animal and die followed by PMS treatment (1,000 I.U. Intergonan) and HCG treatment (250 I.U.). Complement fixation occurred five days after the first hormone application and was followed twelve to 15 days later by another phase of treatment, using Gravigonan (250 I.U. follicle-stimulating hormone (FSH), 500 I.U. HCG, 1 mg oestradiol benzoate in 10 ml serum of swine). Blood samples were continuously drawn during the various phases of treatment and cycle from the above animals as well as from 20 gilts synchronised in the above way and from another 20 untreated gilts. Protein-fixed iodine and copper levels were assessed from those samples. The protein-fixed iodine and copper levels of the blood were significantly reduced (P less than 0.01) by synchronisation, using Suisynchron. None of the two parameters was particularly affected by PMS and HCG treatments. Blood-borne protein-fixed iodine and copper was increased by administration of gonabione. Both parameters went up further during full oestrus at which date they were higher in untreated animals than in synchronised. (Protein-fixed iodine: 3.50/226micrograms/100 ml; P less than 0.01; copper: 0.283/0.234 mg/100 ml; P less than 0.01).


[Histological findings on the uterus of gilts following synchronized ovulation]. Histological studies were undertaken with the view to testing uterus structure and function of gilts following synchronised ovulation by means of different PMSG doses. All proband groups received 500 I.U. HCG. All histomorphological, histochemical, and histometric checks revealed 500 I.U. PMSG to be too low a dosage, whereas the optimum amount was found to be between 750 and 1,000 I.U. PMSG. The uterine glands of all treated animals in all three groups were less favourably developed than those of the untreated controls. The best morphologico-histochemical pattern was observed following administration of 750 I.U. PMSG.


[Non-steroid Suisychron-Premix--its impact on reproduction of gilts in relation to the phase of the sexual cycle]. The action of Suisynchron-Prämix in its dependence on a given phase in the sexual cycle of gilts was studied with the view to improving effectiveness of its administration. A dose of gilts was studied with the view to improving effectiveness of its administration. A dose of 5 g/die was fed to the animals over 20 days, beginning with the third, eleventh, and 16th days of the cycle. The preparation was not fed to the controls. Brood sows in heat were selected and inseminated in compliance with the valid code of practice. Practice-oriented experiments then were conducted and elucidated the effect of Suisynchron upon the reproduction function of the sows concerned. The biologcial and economic effectiveness resulting from the use of Suisynchron, a nonsteroid preparation, was found to depend directly on the phase of the sexual cycle of swine, The regular course of sexual processes in swine is accompanied by characteristic changes, and sexual functions may be either intensified or weakened by administration of Suisynchron-Prämix.


[Possibilities in the use of anabolic substances as veterinary drugs for metaphylaxis and biotechnology]. An account is given of the latest developments and trends in using anabolic preparations in animal husbandry. The development of veterinary medicaments for metaphylaxis and biological engineering in industrialised animal husbandry is explained and substantiated, with reference being made to protein-anabolic and growth-stimulating properties. However, full observation of and compliance with codes and bylaws on drug and food production will depend strongly on efficient interdisciplinary socialist teamworks.


[Exfoliative cytology of nipple discharge (author's transl)]. Due to the exfoliative cytology of the nipple discharge, intracanalicular proliferations within the millimeter range can be detected. Accordingly, this diagnostic method is firmly integrated in the mammary early diagnostics. Within the period reported on so far 3420 women with a pathological secretion could be examined. In 1669 cases there was a bilateral secretion. Thus, altogether 5089 mammae could be cytologically explored. Mammary cytodiagnostics was centered on the bleeding mamma, the myotheliae and galactophoritis. In addition exfoliative cytology can also yield some other diagnostic results, such as the detection of fungiproved in respective cultures. Galactography which was performed in 1053 cases, may be considered to be the only diagnostic consequence resulting from a pathological cytotest. Every 6th galactogram had to be judged as being pathological, and every 5th galactogram histologically clarified revealed a malignant or premalignant proliferation. Accordingly, women manifesting a pathological secretion should be regarded as risk-patients.


Diurnal variations in intensity of symptoms in patients of different diagnostic groups. Consecutive patients, regardless of diagnostic group, upon admission to the Department of Psychiatry, University of Umea, were asked to fill in a special form about diurnal variation in their symptomatology and sleep disturbances. Three trained psychiatrists separately reviewed the records and completed a second form comprising information about diagnosis and certain background factors. Among the background factors only age and ongoing antidepressive medication were found to be of importance. In the study no strong consistency in the symptom complex of early awakening, experience of symptomatology as worst in the morning, and a feeling of relief in the afternoon was found. However, the experience of symptomatology as a whole, anxiety and inhibition as being worst in the morning is significantly more common in patients with unipolar and bipolar affective disorders but is also common in all diagnostic subgroups and seems in no way to be specific for depressive disorders. Furthermore, the phenomenon of diurnal variation of symptomatology was not found to be of diagnostic help and should be employed very cautiously as a basis in pathogenic consideration.


Description of a technique for the analysis of antineuraminidase antibodies oriented to H2N2 and H3N2 influenza virus strains. A method for the analysis of mixtures of antineuraminidase (AN) antibodies oriented to antigenically different neuraminidases of H2N2 and H3N2 influenza virus strains is described. The method is based on the finding that the different classes of AN antibodies give different titer ratios when reacting with the recombinants A/Bel/42 (H0)-A/Sing/1/57 (N2) and A/equine/Prague/1/56 (Heg1)-A/Hong Kong/1/68 (N2) and a mixture of both recombinants. These titer ratios are determined under the experimental conditions of the photometric ACU (antibody concentration unit) method for a series of reference sera and test sera. By mathematical treatment of these data an exact analysis of AN antibodies of test sera is achieved. This paper gives a detailed description of the conduct of the test and summarizes the results of experiments to designed to assess the accuracy of analysis.


[Age and local changes in the architectonics of the lymphatic bed of the wall of the jejunum and ileum in normal rabbits]. Age and local changes of the lymphatic bed have been studied in the jejunum and ileum preparations obtained from 100 rabbits. The architectonics of the bed is stated to undergo some age and local changes. In fetuses and two-week-old rabbits, chyle sinuses are narrow and all flow into the capillaries of the tela submucosa. In one-month-old animals, the mucosal capillary network is developing and the chyle sinuses flow into it. In six--ten-month-old rabbits, intestinal bed is already formed, the chyle sinuses are wide. Aged changes are represented by a considerably rarefied networks, decrease in the chyle sinuses caliber, deformation of the sinus, capillary and vessel walls. In the jejunum, unlike the ileum, the chyle sinuses are of larger size and have more complex architectonics in the capillary networks of the mucosa and tela submucosa.


[Cholinesterase activity in transplantable tissue cultures]. Comparative cytochemical investigation was undertaken to study cholinesterase activity in reinactivated cell strains: Hela, RH, in hamster fibroblasts and in mouse ependymoblastoma. In the cells of ependymoblastoma, butyrilcholinesterase was predominantly discovered. Dynamics of the enzyme activity was inversely proportional to the rate of cell division. Its maximal activity was related to the stage of cellular monolayer formation. Acetylcholinesterase activity in the mouse ependymoblastoma culture was not great.


[Homo- and heterotopic choriocarcinoma in 6- and 18-month-old infants]. Clinical and morphological observations of choriocarcinoma in infants of 6 and 18 months localized in the right lateral ventricle of the brain and in the sacralcoccygeal region are presented. The grounds for verification of the heterotopic tumor include the presence of one large degenerating node in the sacral-coccygeal region and the absence of neoplasia in the central nervous system, its typical histological characteristics corresponding to choriocarcinoma as well as extensive metastasizing along the lymph tracts. The dysontogenetic nature of the tumor is suggested. The difficulties of the clinical and histological diagnosis are due to the rarity of choriocarcinoma and heterotopic localization.


Eye injuries from exploding car batteries. Explosions of car batteries have become a frequent cause of eye injuries. A total of 62 patients were seen in our clinic during eight consecutive months. A wide range of injuries were discovered that extended from the anterior segment to the retina. The most common injuries were acid-induced conjunctivitis, corneal abrasions, and iridocyclitis. Hospitalization and surgery were required in some cases.


Application of the egg-bit technique in poultry-disease research. Experiments were carried out to determine the usefulness of the egg-bit technique (EBT) in poultry virus research with various agents available on the market. The EBT was found satisfactory for propagating 9 strains of avian influenza virus of different antigenic groups, with only minor differences resulting when the procedure used two sources of gelatin, presence or absence of CaCl2, or commercial or specific-pathogen-free embryos.


A quantitative study of pinocytosis and intracellular proteolysis in rat peritoneal macrophages. A method for the culture of rat peritoneal macrophages in vitro is described, in which pinocytic uptake of colloidal [198 Au]gold, 125I--labelled poly(vinylpyrrolidone) and [14C]sucrose proceeds at contant and fairly reproducible rates for several hours. The rat of uptake of colloidal [198 Au]gold, which wxhibited some inter-batch variation, was approx. 100 times that of the other two substrates. Colloidal gold did not affect the rate of uptake of 125I-labelled poly(vinylpyrrolidone) and therefore its own high rate of uptake could not be attributed to a stimulation of the formation of pinocytic vesicles. It conclude that uptake of collodial gold is highly dependent on adsorption on binding sites on the plasma membrane. Uptake of formaldehyde-treated 125I-labelled bovine serum albumin was followed by the release of [125I]iodo-L-tyrosine into the culture medium and took place at a rate intermediate between those of collodial [198Au]gold and the other two non-digestible substrates, 125I-labelled poly(vinylpyrrolidone) and [14C]sucrose.


Stabilization and solubilization of bovine corpus-luteum adenylate cyclase. The effects of guanosine triphosphate, guanosine 5'-[beta,gamma-imido]triphosphate, sodium fluoride and Tris/hydrochloric acid concentration on enzyme activity. 1. Adenylate cyclase of the washed 600g sediment of bovine corpus-luteum homogenate is stimulated by p[NH]ppG (guanosine 5'-[beta,gamma-imido]triphosphate), the imido analogue of GTP, and to a lesser extent by GTP itself. Activation by p[NH]ppG is not reversed by extensive washing before assay, but can, however, be reversed by NaF. 2. Both p[NH]ppG and NaF stabilize the enzyme during incubation at 37 degrees C. NaF also causes an irreversible activation, but only of part of the potentially NaF-activatable adenylate cyclase; there are possibly two components of the adenylate cyclase system, which can be distinguished by their response to NaF. 3. Solubilization of the adenylate cyclase activity in the 600g sediment, by using the non-ionic detergent Lubrol-PX, gave variable yields. A relationship between the magnitude of NaF stimulation of the 600g-sediment enzyme and the yield of soluble activity derived from the sediment was recognized. The results suggest that the pre-existing state of the enzyme complex in vivo is reflected by the response in vitro to NaF and may determine the success with which activity can be solubilized. 4. The absolute yields of soluble activity could be increased by p[NH]ppG preactivation of the 600g sediment. During the development of the maximally active state by preincubation with p[NH]ppG the enzyme passes through a stage in which Lubrol solubilization is increased, but the maximally active state is itself less amenable to solubilization. p[NH]ppG activation causes the appearance of NaF-inhibited states, which appear to be preferentially solubilized by Lubrol-PX.


Subcellular distribution of pyruvate (glyoxylate) aminotransferases in rat liver. The distribution of pyruvate (glyoxylate) aminotransferases in the particulate fraction of rat liver homogenates was examined by centrifugation in a sucrose density graident. Aminotransferase activities towards serine, phenylalanine and histidine with pyruvate and those towards phenylalanine and histidine with glyoxylate were nearly identically distributed. Some 50-55% of the particulate activity was localized in the peroxisomes and the remainder in the mitochondria. Most of alanine-glyoxylate aminotransferase activity was localized in the mitochondria, with some activity in the peroxisomes. Glucagon injection resulted in increases of these enzyme activities in the mitochondria, but not in the peroxisomes.


Hepatic triglyceride accumulation and the ethanol physical withdrawal syndrome in mice. Physical dependence on ethanol was induced in TO strain mice by chronic administration of ethanol by inhalation. The severity of the behavioral syndrome of withdrawal from ethanol was quantified by a subjective scoring method. During the chronic administration of ethanol, triglycerides accumulated in livers of male or female mice with a time course similar to that of the induction of physical dependence. When ethanol was withdrawn from adult or weaning dependent mice, a relationship was observed between the decline of triglyceride concentrations in liver and the duration of the ethanol withdrawal syndrome. The addition of DL-carnitine (7% w/w) to diet during the administration of ethanol markedly inhibited the accumulation of triglycerides, and significantly reduced the intensity of the ethanol withdrawal syndrome. Administration of carbon tetrachloride ((1.3 ml/kg i.p.), however, although augmenting hepatic triglyceride accumulation, had no significant effect on the withdrawal syndrome. The results are interpreted as suggesting either that ethanol-induced liver dysfunction plays a part in dependence, or, more likely, that triglyceride accumulation reflects an ethanol-induced metabolic disorder which is itself related to the induction of dependence.


Neonatal electrocardiographic changes in relation to cardiotocographic changes. Electrocardiograms (ECG) were made immediately after delivery in 28 infants with normal cardiotocographic patterns in labour and in 29 with abnormal patterns. Changes in the S-T segment and T-wave were quantified in a scoring system. High and peaked T-waves (high T/QRS-ratios) were significantly more common in the group with abnormal cardiotocographic patterns. The infants with ECG-changes also had significantly lower Apgar scores and more neonatal complications. The immediate postpartum ECG seems to be a useful indication of the accumulated hypoxic stress to which the child has been subjected in labour.


The presence of an aldehyde dehydrogenase inhibitor in animal diets and its effects on the experimental results in alcohol studies. 1. The effects of chronic ethanol administration on the metabolism of ethanol and acetaldehyde were studied in rats fed on a commercial diet containing an aldehyde dehydrogenase inhibitor, calcium cyanamide (calcium carbimide), as a contaminant in the calcinated bone-meal fraction of the diet. 2. Rats given an ethanol solution (150 ml/l) for 3 months and fed on a diet containing calcinated bone meal showed two times higher activity of the low-Km acetaldehyde dehydrogenase in the liver, 26% higher rate of ethanol elimination, and two to three times lower acetaldehyde levels in blood during ethanol elimination compared with control rats pair-fed on the same diet. 3. The results obtained from the ethanol-treated rats were similar to those obtained in experiments on control rats fed on diets not containing calcinated bone meal. 4. Experiments performed in vitro and in vivo on the inhibition of the acetaldehyde metabolism by cyanamide suggested that the apparent effects of chronic ethanol intake were really caused by calcium cyanamide in the diet.


The hypercholesterolaemic effect of caffeine in rats fed on diets with and without supplementary cholesterol. 1. The effect of caffeine (2.5 g/kg diet) on lipid metabolism was examined in rats fed on a stock (low-cholesterol) diet or on a cholesterol plus cholic acid-supplemented (high-cholesterol) semi-synthetic diet. 2. When caffeine was induced in the stock diet fed to rats for 7 d, there was a moderate but significant increase in the concentration of serum cholesterol compared to the levels observed in the control rats. This change can be accounted for by the increase that was observed in the rate of cholesterogenesis in the liver. 3. After 25 d of caffeine in the stock diet, hepatic cholesterogenesis was still increased but the concentration of serum cholesterol was now the same as in the control rats. During the experimental period there was a progressive increase in the faecal excretion of neutral sterols in the rats receiving caffeine. 4. When caffeine was added to a cholesterol plus cholic acid-supplemented diet, there was a marked increase in the concentration of serum cholesterol but hepatic cholesterogenesis was now reduced. 5. Caffeine in the high-cholesterol diet appeared to delay, but probably did not reduce, the absorption of an oral dose of radio-labelled cholesterol. This conclusion was confirmed using rats which had not previously received either caffeine or cholesterol in the diet. 6. When the effect of caffeine in the high-cholesterol diet was investigated during a 24 h period, an exacerbation of the hypercholesterolaemia was seen only at certain times. 7. After a 4-month period of feeding rats on the caffeine-supplemented high-cholesterol diet, histological examination did not detect any damage to the heart and aorta. 8. The metabolic regulations involved in the effects of caffeine in the two diets are discussed and the relevance of the present results to observations made with human subjects is considered.


Differences in the binding of methylated albumin to non-replicating, replicating and denatured DNA from Ehrlich ascites cells. The binding of methylated albumin to DNA, the basis of the chromatography on columns of kieselguhr coated with methylated albumin (MAK chromatography), was investigated. Scatchard plots revealed only one mode of interaction with fully double-stranded DNA. The complexes should be completely dissociated by raising the NaCl concentration of the solution to 0.8 M, indicating a binding by electrostatic attraction between the oppositely charged protein and DNA molecules. In complexes with denatured and partially single-stranded replicating DNA an additional kind of binding was found which made these complexes more stable against salt dissociation. These secondary interactions were stronger at 23 degrees C than at 0 degrees C and could be weakened by the addition of 6 M urea. It was therefore concluded that apolar forces were involved in these interactions.


Enzymic activities of endo-1,4-beta-D-glucanases purified from Trichoderma viride. Endoglucanases II, III and IV (EC 3.2.1.4) from Trichoderma viride are highly active in degrading CM-cellulose or phosphoric acid swollen cellulose, and only slightly active on Avicel. The specific activities of the endoglucanases increase with the length of the cellooligosaccharide substrates. By rate and product analyses using high pressure liquid chromatography the mode of action of Endoglucanase III was differentiated from that of Endoglucanases II and IV. Endoglucanase III has a low affinity for cellobiose, reacts rapidly with cellotriose, and gradually increases in reactivity with cellooligosaccharides as degree of polymerization increases from four to six. In addition to cleaving internal glycosidic bonds of polymeric substrates, it preferentially cleaves cellobiosyl units from the non-reducing end of oligosaccharides. The cellobiosyl units are often, under initial reaction conditions, transferred to the substrate-acceptor. Endoglucanases II and IV show a preference for internal glycosidic bonds of cellooligosaccharides. The soluble products from the initial action of Endoglucanases II and IV on swollen cellulose are glucose, cellobiose, and cellotriose, which are slowly converted to glucose and some cellobiose.


Characterization of endo-1,4-beta-D-glucanases purified from Trichoderma viride. Four electrophoretically distinct endo-1,4-beta-D-glucanases (EC 3.2.1.4) from Trichoderma viride have been identified and named as isozymes, Endoglucanases I, II, III and IV, according to their electrophoretic mobilities on polyacrylamide gels. Endoglucanases II, III and IV, the homogeneity of each of which was established by discontinuous gel electrophoresis and ultracentrifugation, had specific activities on CM-cellulose of 1010, 60 and 250 specific fluidity units/mg protein, respectively. These enzymes have similar pH optima (pH 4.0-4.5) and are labile at pH values greater than 8.0. The endoglucanases are high in acidic and hydroxylated amino acids and glycine, but low in basic amino acids. Values of 12.0, 10.3 and 13.1 have been determined for the epsilon 1%280 of purified Endoglucanases II, III and IV, respectively. Sedimentation equilibrium analysis has established the molecular weights of Endoglucanases II, III and IV to be 37 200, 52 000 and 49 500, respectively. The three endoglucanases contain mannose, galactose, glucose and glucosamine. Mannose is the principal neutral sugar in each enzyme. Endoglucanase II is distinguished by its low carbohydrate content, 4.5% (w/w), compared to Endoglucanases III and IV which contain 15.0% and 15.2% carbohydrate, respectively.


3-Hexulosephosphate synthase from Methylomonas aminofaciens 77a. Purification, properties and kinetics. 3-Hexulosephosphate synthase (D-arabino-3-hexulose 6-phosphate formaldehyde lyase) was purified from an obligate methylotroph, Methylomonas aminofaciens, to homogeneity as judged by polyacrylamide gel electrophoresis and analytical ultracentrifugation. The molecular weight was determined to be 45 000-47 000 by sedimentation velocity and gel filtration. The enzyme appears to be composed of two identical subunits (Mr = 23 000). A bivalent cation is required for the activation and stabilization of the enzyme. The enzyme is specific for formaldehyde and D-ribulose 5-phosphate. The optimum pH is 8.0 (isoelectric point, pH 5.1) and the optimum temperature is 45 degrees C. Initial velocity studies are consistent with a sequential mechanism. The Michaelis constants are 0.29 mM for formaldehyde and 0.059 mM for D-ribulose 5-phosphate.


Porphyrin biosynthesis: immobilized enzymes and ligands. VI. Studies on succinyl CoA synthetase from cultured soya bean cells. Soybean callus succinyl CoA synthetase (succinate: CoA ligase, (ADP-forming), EC 6.2.1.5), has been chemically bound to Sepharose 4B and some of its properties have been studied. The optimal conditions for binding have been determined. The immobilized enzyme retained 48% of the activity of the soluble enzyme and the coupling yield amounted to 50%. Sepharose-succinyl CoA synthetase can be stored at 4 degrees C for periods up to 90 days with only 25% loss of activity; it can also be repeatedly used without alteration of its enzymic activity. The complex showed enhanced thermal stability; pH optimum was between 7.0 and 8.0 for the bound enzyme, and 8.0 for the free enzyme. A general decrease in the Michaelis-Menten constants for the different substrates of the insoluble enzyme, as compared with values obtained for the free enzyme, was found. Plots of the rate product formation against ATP concentration changed from sigmoideal for the soluble succinyl CoA synthetase to hyperbolic for the immobilized enzyme.


The use of a correlated binomial model for the analysis of certain toxicological experiments. In certain toxicological experiments with laboratory animals, the outcome of interest is the occurrence of dead or malformed fetuses in a litter. Previous investigations have shown that the simple one-parameter binomial and Poisson models generally provide poor fits to this type of binary data. In this paper, a type of correlated binomial model is proposed for use in this situation. First, the model is described in detail and is compared to a beta-binomial model proposed by Williams (1975). These two-parameter models are then contrasted for goodness of fit to some real-life data. Finally, numerical examples are given in which likelihood ratio tests based on these models are employed to assess the significance of treatment-control differences.


Studies related to the metabolism of anabolic steroids in the horse: a gas chromatographic mass spectrometric method to confirm the administration of 19-nortestosterone or its esters to horses. A method is described to confirm the presence of 19-nortestosterone metabolites in urine after the administration of veterinary preparations of this anabolic steroid to horses. The method is based upon the detection, by gas chromatography mass spectrometry or selected ion monitoring, of an isomer of estrane-3,17-diol in the urine.


[Effect of preliminary administration of allogenic cells on transplantation immunity in mice receiving cyclophosphamide]. It was shown that injection of 1 X 10(8) spleen cells from C57Bl/6 mice to CBA mice one day before the injection of cyclophosphamide (CY) helped the take of 2 X 10(7) allogeneic or semiallogeneic cells (injected for the second time 3 to 6 hours after C)). Criterion of survival is the ability of the donor cells to produce antibodies to sheep red blood cells in the recipients tolerant of this antigen. Injection of 1 X 10(8) allogeneic cells two days before CY produces no protective effect. Killer-cells proved to appear on the second day after the immunization with allogeneic cells; their peak was reached on the 5th day. The data obtained suggest that CY eliminated the recipients' lymphocytes, which responded to the transplantation antigens, whereas the killer-cells already formed were stable to the CY action.


[Mechanisms of the reversibility of the stathmokinetic response induced by suboptimal temperature]. Autoradiographic research with leucine-3H showed that the reversibility of the statmokinetic reaction induced by the suboptimal temperature (21 degrees C) did not require any additional protein synthesis and therefore was not connected with the neoformation of the microtubules. Normalization of the mitotic rate was delayed in the presence of copper ions preventing polymerization of microtubules. These data suggest that repolymerization of microtubules subunits was the principal way of mitosis recovery after the action of suboptimal temperature.


[Effect of nerve growth factor and amizil on the viability and metabolism of cultured spinal ganglia]. Spinal ganlia of a 9-day chick embryo were cultivated by the method of "floating rafts" in common medium (control) and in the medium containing amizyl (100 microgram/ml) or a neuregrowth factor (50 microgram/ml). With the action of amizyl there proved to be an increase in the number of surviving neurons; the majority of these neurons contained monoaminoxidase; there was a rise of NAD-diaphorase activity, and, to a lesser extent, of lactic dehydrogenase and isocitric dehydrogenase activities. The neurogrowth factor caused an increase in the number of nerve cells with acetylcholinesterase; there was an elevation of NAD-diaphorase and some rise of malic dehydrogenase activities; the activity of lactic dehydrogenase became maximal; as to succinic dehydrogenase--its activity was somewhat suppressed.


Separation and functional characterization of human neutrophil subpopulations. Human neutrophils have been considered to be a functionally homogeneous population of cells. We have developed a density sedimentation technique for separation of neutrophils into two populations based on their ability to form rosettes with IgG-coated human erythrocytes (7SEA). Under the experimental conditions 80% +/- 4.3% of normal human peripheral blood neutrophilis form rosettes. Functionally rosette-forming neutrophils are more adherent to nylon wool, able to phagocytize more 14C-labeled Staphylococcus aureus, more efficient in killing S. aureus, and more responsive to endotoxin-activated human serum in a 51-cr chemotaxis assay that the non-rosette forming neutrophils. However, there is no difference among neutrophil subpopulations' ability to phagocytize latex particles. Paired samples of exudate neutrophils from cutaneous abscess fluid and peripheral neutrophils from three patients were investigated for their ability to form 7SEA rosettes. In each case exudate neutrophils contained greater than 96% rosette-forming neutrophils, whereas peripheral blood contained the normal 80% ( less than 0.01). Thus we show that peripheral blood contains at least two distinct populations of neutrophils. However, an essentially homogeneous neutrophil population is present in cutaneous exudate fluid.


Does "afternoon diabetes" predict diabetes? Twenty-eight men were given morning and afternoon oral glucose tolerance tests in 1969 and again in 1975. According to British Diabetic Association criteria all 28 had normal morning values in 1969 but seven had "afternoon diabetes". Four men had diabetic values in the morning in 1975 but only two of these had had afternoon diabetes in 1969. Better prediction of subsequent diabetes was obtained by calculating the area under the morning glucose tolerance curve in 1969. All four men who progressed to diabetes had areas exceeding 1000 units, which distinguished them absolutely from the other 24. They also tended to be more obese, but this was less predictive of subsequent diabetes.


Idiopathic thrombocytopenic purpura in Hodgkin's disease: a rare occurrence of no prognostic significance. Six patients with Hodgkin's disease who developed idiopathic thrombocytopenic purpura (ITP) are presented. In contrast to prior cases reported in the literature, none of these patients developed recurrent Hodgkin's disease at the time of or subsequent to their thrombocytopenia, with follow-up data available up to 10 years. ITP does not necessarily imply recurrent or active malignancy.


Oscillating contractions in protoplasmic strands of Physarum: effects of external Ca++-depletion and Ca++-antagonistic drugs on intrinsic contraction automaticity. 1. The minimal requirement of external Ca-concentration for continuance of contraction activity lies in the range of 10(-4) M. 2. As in mammalian smooth muscle, the Ca antagonistic drugs verapamil and D 600 (5.10(-4) M) suppress the minute-rhythms. The action of the drugs is inhibited by 5mM Ca, La or Mn. De novo generation of contraction automaticity is not inhibited by external Ca depletion or by Ca antagonists. 3. It is concluded that rhythmical Ca fluxes across the cortical plasmalemma are not a precondition for triggering continuance or de novo generation of contraction automaticity.


Sex differences in averaged visual evoked potentials during food intake in rats. Averaged visual evoked potentials (AVEP's) were recorded in unrestrained male and female rats during alert wakefulness (baseline condition), feeding and drinking. The rats had been deprived of food and water for 23 hr prior to testing. Peak-to-peak amplitude of the AVEP's was measured between the slow negative and the preceding positive component. Under baseline conditions females had higher amplitude AVEP's than males. It is suggested that sex differences in adrenal steroid secretion could account for this observation. During feeding and drinking, AVEP's increased significantly in amplitude in both sexes, and high-voltage, slow EEG activity was observed to appear in most animals. During feeding, these increases in AVEO amplitude were significantly larger in males than in females. These findings support recent views on sex differences in the regulation of feeding behavior.


Gene mapping by fluorescent in situ hybridization. We describe a new method for the mapping of mammalian genes, utilizing in situ hybridization of mRNA to DNA of chromosomes. It involves the hydrogen bonding of the polyadenylic acid at the 3' end of hybridized mRNA to the polyuridylic acid tail of a highly fluorescent latex microsphere. The resultant double hybrid can be visualized by fluorescence microscopy. The chromosomal localization of human alpha + beta globin genes has been explored by this method. Our data point ot the long arms of chromosomes 4 and 5 as the loci for the human globin genes.


Ordered arrays of intramembrane particles on the surface of fish gills. Freeze fracture of euryhaline fish gill epithelium reveals large smooth surfaces with intramembrane particles or corresponding pits often aggregated into hexagonal arrays. They belong to the outer plasma membranes of respiratory cells, the main surface of contact between the internal and external milieu of the fish. Possible functions for the arrays are discussed in relation to the fine structure of the respiratory cell.


Enhancement of movement of mouse peritoneal macrophages caused by colchicine treatment. Colchicine in the concentration range of 10(-8) to 10(-4)M enhances the movement of mouse peritoneal macrophages while at higher concentrations it inhibits movement. Maximum movement is observed in the concentration range of 10(-6) to 10(-4)M.


Milk fat globule membranes devoid of intramembranous particles. When isolated milk fat globule membranes from bovine, human, and murine (rat) milk were examined by freeze-fracturing most of the membrane faces were devoid of membrane-intercalated particles whereas a minor portion showed relatively few particles, either in clusters or in apparent random distribution. A reduced particle density was also noted in membranes of intra-alveolar milk fat globules of cows and rats, in contrast to high particle densities in the apical plasma membrane of lactating epithelial cells. The observations suggest that certain membrane constituents recognized as intramembranous particles either are displaced from the region of the apical surface of the mammary epithelial cell which is involved in milk fat globule budding or are dislocated and rearranged during the budding process.


Determinants of growth in patients with ventricular septal defect. Growth status was studied in 1210 patients with ventricular septal defects (VSD) participating in the Joint Study on the Natural History of Congenital Heart Defects. A total of 793 patients were managed medically and 194 underwent successful surgical repair. Admission measurements revealed height and weight status to be subnormal and related to physiological severity and age. Successful repair resulted in significant increase in weight but not height. Medical therapy was associated with little change in the subnormal growth pattern. Multivariate analysis revealed determinants of admission height for all patients with VSD to be presence of extracardiac anomalies, birthweight below 2.5 kg, admission age greater than 2 years, history of heart failure and pulmonary/systemic pressure ratio. The same determinants were noted for admission weight with addition of pulmonary/systemic flow ratio. It is conluded that the severe growth disturbance in patients with VSD is only in part due to abnormal postnatal hemodynamics. Intrauterine and genetic factors and low birthweight also play a role and help explain the incomplete growth response after successful surgery.


Radioimmunoassay of creatine kinase isoenzymes in human serum: isoenzyme MM. Measurement of the mass concentration of serum enzymes by radioimmunoassay provides direct quantitation of specific isoenzymes and may be less subject to some of the limitations of traditional assay procedures for enzymes. We describe the development of a sensitive and specific radioimmunoassay for the muscle isoenzyme of creatine kinase, CM-MM, in human serum. CK-MM, purified from human skeletal muscle, was used to raise high-titer antisera and for iodination by the Chloramine T method. The radioimmunoassay required 50 microliter of sample, utilized a double-antibody separation method, and was completed in 24 h. Cross reactivity with CK-BB was virtually zero, 3--17% with CK-MB. The mass concentration of CK-MM in the serum of healthy subjects ranged from 36 to 1668 microgram/liter and correlated closely with total CK enzymatic activity. Serum concentrations of CK-MM from casually selected patients correlated less well with total CK enzymatic activity, suggesting the existence of other CK isoenzymes or the presence of inactive forms.


Radioimmunoassay of creatine kinase isoenzymes in human serum: isoenzyme BB. We describe a sensitive, specific radioimmunoassay for the BB isoenzyme of creatine kinase (CK-BB) in serum. A sequential saturation assay was used to achieve sufficient sensitivity to detect the isoenzyme in 100-microliter serum samples of all healthy persons and patients tested. Bound and free antigen were separated by a second antibody system. Large excesses of purified isoenzyme MM did not react in the assay. Cross reactivity of two preparations of CK-MB was only 1 to 7+. The 95th percentile of serum CK-BB in 208 healthy adults was 6.2 microgram/liter. Within-assay and between-assay precision ranged from 5.5 to 11.9% and 9.7 to 13.6%, respectively.


Parathyrin radioimmunoassay: diagnostic utility of antisera produced against carboxyl-terminal fragments of the hormone from the human. Antisera directed toward the carboxyl-terminal region of human parathyrin (parathyroid hormone), for use in daignostically applicable radioimmunoassays of the hormone in serum, are scarce, largely because of the lack of suitable immunogens of human origin. We produced four antisera in goats and guinea pigs by immunization with recently discovered carboxyl-terminal fragments of human parathyrin extracted from parathyroid tumors. Here, we report results of radioimmunoassays of nearly 200 normal and pathological sera with one of these antisera; we observed almost complete differentiation between concentrations of parathyrin in serum of healthy normal subjects and patients with primary, secondary (due to chronic renal failure), or "ectopic" hyperparathyroidism (due to nonparathyroid cancer). The availability of a new immunogen should now make possible the deliberate production of large quantities of diagnostically applicable parathyrin antisera directed toward the carboxyl-terminal region of human parathyrin. This should, in turn, lead to more widespread availability of this useful radioimmunoassay.


The pharmacology of thebaine in the chronic spinal dog. The effects of thebaine were studied in nondependent and morphine-dependent chronic spinal dogs as well as in chronic spinal dogs receiving thebaine chronically. Thebaine did not produce any morphine-like effects in nondependent dogs nor did it precipitate an abstinence syndrome in morphine-dependent dogs. Large doses of naltrexone precipitated a mild abstinence syndrome in dogs receiving thebaine chronically; however, no withdrawal abstinence syndrome was observed after abrupt withdrawal. Thebaine is not a morphine-like drug, nor does it produce appreciable physical dependence in the dog.


Dehydrogenase activity of the pineal gland during the estrous cycle. An investigation of the relationship of succinic dehydrogenase (SDH) and lactate dehydrogenase (LDH) in the pineal gland of the adult female albino rats was carried out during seven definitive stages of the estrous cycle: Early Estrus (E1), Late Estrus (E2), Metestrus (ME), Early Diestrus (D1), Late Diestrus (D2), Preproestrus (PPE) and Proestrus (PE). Controlled housing conditions were maintained on 7:00 A.M. to 7:00 P.M. light-dark cycle. The animals were necropsied at precise times after the onset of estrus and subsequent stages of the cycle. Comparative histochemical and biochemical analysis revealed the following for SDH activity: E2 greater than E1 greater than D2 greater than D1 greater than PPE greater than PE greater than ME; and for LDH activity: PE greater than E2 greater than ME greater than D1 greater than PPE greater than E1 greater than D2. Interrelationships are postulated between pineal SDH, GOT, GPT, and estrogen concentrations in the rat.


Vaginal stone in a male pseudohermaphrodite. A rare case of vaginal stone in a male pseudohermaphrodite who had undergone plastic surgery for external genitalia 16 years ago is presented. It is recommended to remove the entire wall of vagina musculina at the time of plastic repair for hypospadias in a male pseudohermaphrodite.


Increased antagonist potency of naloxone caused by morphine pretreatment in mice. Using the writhing test in mice, it was shown that pretreatment with a single dose of morphine hydrochloride given 3 h previously caused a marked increase in the antagonistic effect of naloxone without any change in the antinociceptive action of morphine itself. It was shown that when mice were pretreated with different doses of either morphine alone, or in combination with naloxone, so that each treatment produced the same antinociceptive effect, the increase in naloxone potency was proportional only to the antinociceptive effect of the pretreatment and not to the total dose of morphine present. It was also found that the concurrent administration of naloxone plus morphine prevented the development of "acute dependence" to morphine, as measured by the jumping reaction after challenge with naloxone.


Ovarian serotonin content in relation to ovulation. In the mature cyclic female rat, analysis for ovarian serotonin content reveals comparatively high serotonin content. Fluctuation of serotonin content was observed; peak for serotonin was observed at estrus. In gonadotropin-treated immature rats, there was no detected ovarian serotonin using this procedure. It was concluded that ovaries from gonadotropin-treated immature rats are physiologically different from ovaries taken from mature cyclic rats.


[Studies on the measurements of urinary pregnanediol by radioimmunoassay (author's transl)]. A simple and practical radioimmunoassy (RIA) for pregnanediol without any pretreatment of urine specimens such as hydrolysis and/or extraction was developed. Antisera were produced by immunizing rabbits with pregnanediol-3-glucuronide (P-diol-3-G)-BSA conjugates and P-diol-3-hemisuccinate-BSA conjugates. The specificities of the antisera were examined by their cross-reactivities with related steroids. Antiserum to P-diol-3-G-BSA was significantly cross-reacted with 20 alpha-OH-P(201%), P-diol(100%), P -diol-3-G (77.3%, where 119% as free P-diol), to a lesser extent with 20 beta-OH-P(17.8%), and did not react with other steroids. Antiserum to P-diol-3-hemisuccinate-BSA, on the other hand, was capable of reacting with P-diol(100%), P-diol-3-G(46.9%), 20 alpha-OH-P(75.2%) and 20beta-OH-P (8.97%), respectively. Antiserum generated against P-diol-3-G-BSA was almost twice as high in its avidity for P-diol-3-G as compared with antiserum raised against P-diol-3-hemisuccinate-BSA. Antiserum to 20alpha-OH-P-3-carboxy-methyl oxime-BSA conjugate was only reactive with 20alpha-OH-P and not with P-diol or P-diol-3-G. From the data presented here, antiserum to P-diol-3-G-BSA with high specificity toward urinary P-diol-3-G was used for the present assays. Ammonium sulfate precipitation was employed for the separation of antibody-bound and free tracer hormones. A direct method in which simply diluted urine was directly applied to RIA and the enzymatic hydrolysis method which requires extraction of P-diol with ether following hydrolysis of urine by beta-glucuronidase were both examined in the present RIA on the same specimens which contained known amounts of 3H-P-diol-3-G. Values obtained by the direct method were in good correlation with those determined by the hydrolysis method (r=0.994 p less than 0.01, Y=1.07X + 0.30). The working range of the assay was from 50pg to 5ng of P-diol per tube, and intra- and interassay precisions were 9.8, 12.5 and 12.9, 16.7%, respectively. Determination of radioimmunoassayable P-diol-3-G in urine was also in good correlation with the values obtained by the currently used analytical procedure, the Klopper-Kambegawa method.


A components method for measuring the impact of a family planning program on birth rates. One of the major goals of family planning programs worldwide has been to reduce the level of fertility in hopes of slowing the rate of natural increase and promoting social and economic development. Such programs have now been in existence for sufficient lengths of time to have had an impact on fertility levels. In general countries with organized family planning programs, marked declines in fertility levels have been observed. The extent to which such declines may be credited to organized programs has not been rigorously measured because an appropriate research methodology has been lacking. This paper describes one method of directly linking declines in fertility levels to the contraceptive protection experienced by a population. The contribution of organized family planning programs is estimated by decomposing the amount of total contraceptive protection into within-program and outside-program sources.


An observational study of caretaking behavior of male and female mice housed together. A time-sampling procedure was employed to examine the caretaking behavior of male mice housed with lactating females and their litters, and to investigate the effects of the male's presence on the behavior of the female. The incidences of 14 categories of behavior were recorded over 20 hr distributed throughout the period of lactation. The development of litters was monitored by regular weighing. The results show, contrary to earlier findings, that males display just as much caretaking behavior as females. Differences in other forms of behavior can be related to the increased nutritional needs of the nursing mothers. The presence of the male seems to have very little influence on the behavior of the female and no effect at all upon the physical development of the litter.


Female mouse maturation: effects of excreted and bladder urine from juvenile and adult males. A sequence of 4 experiments examined the effects of prepubertal and adult males on the sexual maturation of young female house mice. The results support 3 conclusions: (1) the presence of a prepubertal male or of urine from prepubertal males does not affect the timing of sexual maturation in young female house mice; (2) the maturation-accelerating pheromone produced by adult males is present in the bladder urine of intact adult males but is absent from both excreted and bladder urine of castrated males; and (3) young females caged with 7 prepubertal males or with a castrated adult male mature earlier than control females caged alone. Results indicating that the presence of a castrated male leads to earlier sexual maturation of young female mice differ from previous findings. A possible explanation for this contradictory result is based on the ability of young weanling female mice to acclimatize the thermoregulate when separated from the dam and litter-mates. A model for density-feedback population regulation in house mice involving pheromones produced by males and females is presented.


Characterization of seven C-peptide antisera. The plasma C-peptide immunoreactivity (CPR) in 10 normal subjects varied considerably when measured with different antisera in parallel assays. The CPR level correlated with the blank "CPR" value measured in plasma devoid of C-peptide and to a lesser degree with the sensitivity of the standard curves obtained with the individual antisera. Storage of plasma samples at different temperatures and for different lengths of time before the analyses were carried out resulted in further variation in the CPR results. This was caused by a time- and temperature-dependent fall in CPR, which was more pronounced with some antisera than with others. This sensitivity to storage of plasma did not correlate with the antigenic characteristics of the antisera as determined by their reactivity with 11 specific fragments of the C-peptide molecule. The contribution of human proinsulin to the CPR concentration relative in normal subjects was considered to be negligible even though the relative immunoreactivity of human proinsulin and C-peptide ranged from 11 to 143 per cent among these antisera. These results suggest that differences in C-peptide antisera are a major reason for the variation in the concentration of circulating CPR as measured in different C-peptide immunoassays.


Neonatal stimulation, maternal behavior, and accelerated maturation in BALB/c mice. An experiment was conducted to examine the effects of neonatal handling and hypothermia on infant physical maturation and growth and on maternal behavior in BALB/c mice. Stringent methodological and statistical controls were employed for experimenter bias and litter effects. No evidence appeared to support claims that neonatal stimulation leads to accelerated physical maturation. Alterations in maternal behavior patterns were found only immediately following hypothermia of pups. Maternal behavior did not correlate highly with offspring maturational rates, but maternal weight and age and pup birth weight were highly predictive of infant physical maturation.


[The course of hypertrophic obstructive cardiomyopathy. A preliminary study of 40 cases (author's transl)]. The course of hyperthrophic obstructive cardiomyopathy in 40 patients was studied. The average follow-up of the patients (anamnestic and prospective) was 8 years. The age of the first diagnosis, the age of the first observation and the age of the detection of the first symptom, were considered. A prospective follow-up study was carried out in 31 patients, the average period of observation being 8 years. Functional capacity modifications, symptoms, clinical and electrocardiographic signs and the mortality rate  were analyzed. Functional class unchanged in the most part of the patients; the overall mortality was 16% with high incidence of sudden and unexpected death. From our results it appeared that the prognosis could be related to the old age, heart failure and arrhythmias.


Small intestinal epithelial cell kinetics and protozoal infection in mice. The effects of chronic protozoal infection on small intestinal architecture have been examined in mice, infected with Giardia muris and Hexamita muris. Techniques used were conventional histology, quantitation of intraepithelial lymphocytes, microdissection and measurement of individual villi and crypts, and epithelial cell kinetic studies. The histology of small intestine from infected mice appeared normal apart from the intraepithelial lymphocyte numbers. Mean intraepithelial lymphocyte counts in two groups of uninfected mice were 11.6 and 13.6 per 100 epithelial cells, and in two groups of infected mice were 17.6 and 21.8. Dynamic studies showed that protozoal infection doubled the cell production per crypt per hour from mean values of 6.2, 7.3, and 8.2 in three groups of uninfected animals, to 11.8, 13.4, and 17.1 in groups of chronically infected mice. Cell production per villus was also influenced by protozoal infection, with values of 93, 99, and 101 cells per hr in groups of uninfected animals whereas in infected mice the values were 155, 162, and 180 cells per hr. Although there was no reduction in villus height in the infected animals, radioautography using [3H]thymidine confirmed that the enterocytes moved rapidly up the sides of the villi than was the case for uninfected mice.


[Skin defects in the newborn or fetus as questionable sequelae of amniocentesis in early pregnancy (author's transl)]. In three of 111 infants born after amniocentesis and in one of 5 fetuses aborted after amniocentesis, which could be examined, small skin lesions or scars were observed. Interpretation as sequelae of needle puncture appears possible, but questionable, particularly since similar lesions were observed in infants born after uncomplicated pregnancy without amniocentesis. The differential diagnosis of a focal dermal hypoplasia or aplasia may be considered, which may be hereditary.


Immune-complex mediated colitis in rabbits. An experimental model. An experimental colitis in rabbits is described, following the intravenous injection of preformed immune complexes of human serum albumin (HSA) and anti-HSA into non-sensitised rabbits. Tissue damage was localised to the colon by the Auer technique of inducing local non-specific inflammation, by the rectal instillation of dilute formalin. Formalin alone gave transient changes that reverted to normal within 24 hours. In rabbits given intravenous immune complexes formed in antigen-excess, a severe colitis was initiated, with histological features including mucosal ulceration, mixed inflammatory cell infiltration in the lamina propria, and crypt abscess formation. It is possible that immune-complex damage may be one of the pathogenic mechanisms involved in human ulcerative colitis.


The effect of parathyroid hormone and vitamin D3 metabolites on CaBP synthesis by embryonic duodenum, in vitro. Embryonic chick duodena were incubated, in vitro, for two days in the presence of vitamin D analogues and metabolites and the ability of PTH to potentiate the vitamin-D-dependent synthesis of CaBP measured. PTH potentiated CaBP synthesis in the presence of vitamin D3, 25-OH-D3, 1 alpha-OH-D3, DHT, and 1,25(OH)2D3. The synthesis of CaBP was directly related to the log of the meddium PTH concentration over the range 1 x 10(-8) M to 5 x 10(-7) M.


Utilization review can be a tool for improvement. The battle still rages over how effective PSROs are in conducting patient care review. A competitive win-lose relationship between the PSRO and the hospital to which it has delegated review authority, two PSRO experts caution, will only rebound to the hospital's disadvantage. However, a partnership between the two, they say, will help alleviate some of the problems that are bound to occur.


Bursal fabricii as a peripheral lymphoid organ. Transport of various materials from the anal lips to the bursal lymphoid follicles with reference to its immunological importance. In tracer studies, various particulate and molecular materials applied on the anal lips of chickens were transported to the bursal lumen. From here some of the tracers were further transported into the bursal lymphoid follicles and colloidal carbon even to the bursal stroma. In immunization experiments, Brucella abortus organisms and SRBC were applied on the anal lips of 1-day-old, 4-week-old and 10-week-old chicks on 5 consecutive days. Although Brucella bacteria in tracer studies were not found in the bursal tissue, agglutinin response to Brucella was observed in the 4-week and 10-week groups as early as 3 days after the last application of this antigen. The response to SRBC was strong only in the oldest group of chickens. When stimulated continuously, the agglutinin titres reached their highest values after the 2nd week and then began to decrease to rather low levels. In the youngest age group, the anti-SRBC titres, however, remained low during the whole experimental period, and even in the 4-week group the anti-SRBC response was weak. Bursectomy, carried out at the age of 10 weeks, inhibited the agglutinin response to Brucella but affected only little the response to SRBC. It is concluded that by taking up antigens per anum the chicken possibly gains part of its basic immunity. In this immune response the bursa seems to play an important role at least with some antigens, and thus the bursa may have an immunofunction comparable with that found in peripheral lymphoid organs.


Homocytotropic response of guinea-pigs to antigen extracts from formalized and unformalized Parascaris equorum. A homocytotropic anaphylactic response is described in guinea-pigs sensitized with antigen from Parascaris equorum. This antigen was not sensitive to prolonged storage in formalin. Some adult ascarid extracts prepared from worms that had been stored for 14 years in 10 per cent formalin before use, retained their ability to produce an allergenic response. The antisera withstood heat inactivation at 56 degrees for up to 7 h or treatment with 2-mercaptoethanol and were still capable of eliciting a skin response. The skin response persisted for 96 h after sensitization. The serum of animals sensitized with a single dose of P. equorum antigen remained capable of producing a positive passive cutaneous anaphylactic response up to 245 days after the sensitizing dose. It appears, from the results obtained, that this antibody resembles IgG1, rather than IgE.


[Human catabolism of medium and long chain triglycerides after intravenous infusion]. The metabolic rate of intravenously administered middle-chain and long-chain triglycerides has been tested in metabolically healthy adult subjects. After an infusion period of three hours (infusion rate 0.22 g triglyceride/kg body-weight and hour) the metabolic rate of long-chain triglycerides was 16.4 mg/kg body-weight and hour. The metabolic rate of the middle-chain triglycerides (infusion rate 0.056 g/kg body-weight and hour) administered in a mixture with long-chain triglycerides was slower: 4.8 mg/kg body-weight and hour. Even under these conditions, middle-chain triglycerides increased ketonemia and ketonuria. Furthermore, the acetate level in blood increased significantly. Infusion of lipids have a strong effect on amino-acid levels in blood. The concentration of alanine, leucine, isoleucine are decreased, whereas the concentration of glutamate increases at the same time. Under clinical parameters only a mild leucocytosis has been found. Our data suggest that the metabolic rate of middle-chain triglycerides is according to their low plasma concentration slow, nevertheless, the ketogenesis is significantly higher if middle-chain triglycerides are infused in a mixture with long-chain triglycerides.


[Comparative study on the immunogenicity of plasma substitutes with gelatin base in animal experiments]. The immune response of rabbits and guinea pigs to gelatin and to three commercial plasma substitutes based on gelatin (oxypolygelatin [OPG], di-isocyanate cross-linked gelatin [DCG], modified fluid gelatin [MFG]) has been reevaluated. Passive hemagglutination, passive cutaneous anaphylaxis and active anaphylaxis have been used to detect the response of animals immunized in complete Fruend's adjuvant or in aluminium hydroxide. Marked immune responses were observed with DCG and MFG which were to a large extent specific for the immunizing antigen (i.e. the corresponding chemically modified gelatin). Gelatin and OPG induced weak responses. In contrast to DCG and MFG in systemic anaphylactic shock experiments no anaphylactic shock could be elicited with gelatin and OPG. A limited series of immunizations in guinea pigs of various strains demonstrated that responses to weak immunogens, such as modified gelatins, are markedly influenced by genetic factors.


[Animal experimental studies on liver morphology after intravenous administration of a fat emulsion]. In short infusion series run over a 5day period, a fat emulsion on the basis of soya oil with egg lecithin as emulsifier, has been applied to rabbits by the intravenous route in a dosage of 2 and 4 g/kg body-weight/day. The results changes to the liver morphology have been analysed histologically by light-microscope in a systematic manner; using standard intervals of one to fifteen weeks. The most conspicuous phenomenon of the changed liver morphology is a fatty degeneration of the Kupffer's cells of the liver, whose different degree of severity was determined semiquantitatively by determination of the so-called index of fatty degeneration. The loading of the reticulo-endothelial system (RES) of the liver is very heavy with the higher dose; it is lower with a dose of 2 g fat/kg body-weight. The histological changes recede only slowly. A simultaneous parenteral administration of carbohydrates did not result in any provable inferior loading of the RES.


[Intraportal and intravenous administration of fat emulsions. Comparison of liver morphology in animal experiments]. In short infusion series run over 5 days, a fat emulsion on the basis of soya oil with egg lecithin used as emulsifier has been infused to rabbits by the intravenous and intraportal routes in a dosage of 2 and 4 g/kg bodyweight/day. By a light-microscopic analysis of the liver morphology after one and three weeks, mainly a fatty degeneration of the Kupffer's cells of the liver was found. It was considerably less conspicuous after endo-portal supply than after peripheric-intravenous supply--findings which became more obvious with the higher dose.


Lipid-free glycerol teichoic acids with potent membrane-binding activity. Lipid analysis of several glycerol teichoic acid preparations strongly indicated that covalently bound lipid is not required for spontaneous adsorption of glycerol teichoic acid to erythrocyte membranes. Although fatty acids were detected in each of four batches, none were covalently bound. Chloroform-ether-extracted antigens retained potent erythrocyte membrane-binding activity as measured by passive hemagglutination, even though they were shown to contain less than one fatty acid residue per 4,869 teichoic acid chains. Mild ammonolysis abolished erythrocyte-sensitizing activity in passive hemagglutination, but further studies indicated the loss of activity was due to partial destruction of the polyglycerophosphate backbone and not to the removal of esterified lipid. The amount of hydrolyzed antigen required to produce 100% passive hemagglutination inhibition was between 170 and 330 times the amount required to produce the same result using unhydrolyzed glycerol teichoic acid. The average chain length was reduced from 19.1 to 9.7, 7.4, and 5.1 glycerophosphate residues for antigen samples hydrolyzed for 1, 5, and 16 h, respectively.


Hydrogen peroxide release by rat alveolar macrophages: comparison with blood neutrophils. Hydrogen peroxide release was examined using biochemical and cytochemical techniques in rat alveolar macrophages, at rest and during phagocytosis, and compared with rat blood neutrophils. Using biochemical techniques, alveolar macrophages released small amounts of hydrogen peroxide at rest, and no increase was observed after challenge with opsonized and nonopsonized zymosan particles at several particle-cell ratios (1:1 to 1:1,000). Neutrophils released similar quantities of hydrogen peroxide at rest but showed a 12-fold increase in hydrogen peroxide release following exposure to opsonized zymosan particles. Using cytochemical techniques to localize sites of hydrogen peroxide release, resting neutrophils showed little deposition of reaction product at the cell surface and occasional deposits in endocytotic vesicles. After exposure to latex particles, a dense reaction product was observed between the particle and the cell membrane, indicating significant increases in hydrogen peroxide release at the sites of particle contact with the neutrophil. The resting macrophage displayed a light, uniform precipitation of cerium over the cell surface and lining intracellular channels and endocytotic vesicles and vacuoles. Following particle exposure, there was no significant difference in the density or distribution of reaction product. These findings, together with previous studies of oxidative metabolism, suggest that alveolar macrophages do not release increased quantities of hydrogen peroxide during phagocytosis. In contrast to neutrophils, oxidative-dependent metabolic pathways may not be of primary importance for microbial killing by alveolar macrophages.


Motives for drug use: an application of Cohen's typology. A modified version of Cohen's typology of motives leading to drug use was studied in three private schools, Grades 7 through 12, two colleges, and a random sample of adults in two cities in a rural county located in the Intermountain West. The distribution of motives for the use of 12 categories of drugs was summarized for each population and comparisons made for age and sex differences. Younger respondents reported a higher use of drugs for adventure/curiosity reasons than older respondents. Students using drugs at a college having unusually low drug use showed a higher level of general motive satisfaction than students at a college having average drug use. Females reported a somewhat higher level of general motive satisfaction than males, but the sex differences observed were not consistent with common sex role stereotypes. By applying these motive distributions in a given community or institution, and following Cohen's suggestions for alternatives to drug use, it would be possible to build an effective program aimed at the prevention of drug abuse and related problems.


Ocular immunoglobulins in lepromatous leprosy. Immunoglobulin levels in the ocular fluids have been estimated in normal subjects and lepromatous leprosy patients. In the normal tear, IgA is the major immunoglobulin while IgG is the only immunoglobulin detected in the aqueous humor. The immunoglobulin profiles in the tear and the aqueous humor in normal subjects are different. The mean IgA level in the tears of the lepromatous leprosy group is significantly lower than in the control patients. IgA and IgG levels are raised in the aqueous humor of some leprosy cases who had suffered from uveitis in the past and also in all cases with active endogenous uveitis. Therefore, in lepromatous leprosy the pattern of immunoglobulin alteration in the tear and the aqueous humor is not parallel.


Epidemiology of leprosy in Iran.

Epidemiology of leprosy in Iran. A total of 907 cases of leprosy from two sources, records from Baba-Baghi Leprosarium (709 cases) and Ahar case finding survey (198 cases), have been studied. The main characteristics of the cases are: a) about 50% of all cases are lepromatous leprosy; b) the leprosarium cases are about 2.5 years younger; c) about 70% of all cases are male; and d) the incidence of leprosy shows a steady increase up to 25-30 years of age and levels off thereafter. These and other findings are discussed.


The prevalence and pattern of leprosy in a school survey. The prevalence of leprosy in a school survey conducted in Panaji, India was found to be 5.3 per 1,000 with males predominating. The majority of patients had a single lesion on exposed parts of the body showing the clinical characteristics of tuberculoid leprosy. However, clinical features of indeterminate leprosy were seen in two patients and borderline tuberculoid in a single case. On the other hand, histopathologically, the majority of the patients were classified as having borderline tuberculoid or indeterminate leprosy. A disparity between the clinical and histopathologic diagnosis was evident. This observation emphasizes the importance of studying both the clinical and histopathologic features in deciding the precise status of a patient in the leprosy spectrum.


Time course changes in total lipid content and fatty acid distribution in various tissues of young chickens after feeding rapeseed oil or soybean oil. 1. In chicks fed rations containing 15% rapeseed oil (RSO) or soybean oil (SBO) a gradual increase in the lipid content of heart tissue was found from 1 to 10 days of age. The finding that feeding high levels of RSO to chicks did not cause excessive accumulation of fat in cardiac tissue supports the contention that it is inappropriate to extrapolate from one species to another (e.g. from rats to humans) as has been done by some workers with respects to RSO utilization. 2. Inclusion of RSO in the ration caused an increase in the total omega-9-monounsaturated fatty acid concentration in heart tissue. The increase in C 20:1 fatty acid concentration in heart tissue noted in the RSO-fed chicks suggests beta-oxidation of erucic acid (delta13 22:1) to eicosenoic acid (delta11 20:1). The rapid decrease in the oleic acid (delta9 18:1) concentration of liver tissue of chicks found during the period 1 to 10 days of age suggests the importance of oleic acid as a source of energy for the chicken.


Fine needle aspiration of lymph nodes in leprosy. A study of bacteriologic and morphologic indices. Lymph node aspiration was performed from the inguinal group of lymph nodes in 16 patients having lepromatous or borderline leprosy. In the same group of patients impression smears of excised lymph node and slit smears of the skin were also studied. This study made it obvious that the aspiration biopsy technic gave similar information regarding the Bacteriologic (BI) and Morphologic Indices (MI) as the impression smears of excised lymph nodes. The technic of aspiration being simple and nontraumatic is recommended for the follow-up of patients on antileprosy drugs and in reactional phases.


Transurethral sphincteric stimulation.

Transurethral sphincteric stimulation. Electrical stimulation of the external urethral sphincter is a valuable aid in teaching sphincterotomies and transurethral procedures. It helps accurately locate the external urethral sphincter which should be transected totally during a sphincterotomy and left intact in a transurethral procedure.


The value of routein intravenous urography in acute epididymitis. The urographies of 106 patients with acute epididymitis are reviewed. In 71 cases the urogrpahy was completely normal, while in the remaining 35 patients abnormalities were found. In 25 patients the findings were consistent with hypertrophy of the prostate. More than half of the patients had known urological diseases and had undergone cystoscopy and/or catheterization prior to the onset of acute epididymitis. All 35 patients had demonstrated symptoms that would otherwise have resulted in intravenous urography independent of the epididymitis. No cases of urogenital tuberculosis were found. The authors conclude that routine urography under the age of 50 is not recommended. Over the age of 50 concomitant urological diseases are so common that urography is often indicated, but acute epidiymitis in itself is only a relative indication.


Relationship of San Miguel sea lion virus to other members of the calicivirus group. San Miguel sea lion virus (SMSV) is indistinguishable from vesicular exanthema virus (VEV) and feline calicivirus (FCV) in its morphology and in possessing a single capsid polypeptide with a molecular weight of approximately 65 X 10(3). Neutralization tests readily differentiate the three viruses, but immunodiffusion tests show that SMSV is closely related serologically to VEV but not to FCV. Although the RNAs of the three caliciviruses have similar base compositions, homology tests show that SMSV is closely related to VEV but is not related to FCV. Tryptic peptide maps of the single major polypeptide comprising the capsid of each virus also show that SMSV and VEV are more closely related to each other than to FCV.


Cow's milk intolerance presenting as necrotizing enterocolitis. Case history is reported of an infant who was changed from breast to bottle feeding at fourth day of life, who on ninth day developed bilious vomiting and bloody diarrhoea. Later gas was found in colonic wall on X-ray. Necrotizing enterocolitis was diagnosed and infant managed with total intravenous alimentation. On recovery although she tolerated breast milk, return to cow's milk formula lead to return of bloody diarrhoea. Subsequently she thrived on a cow's milk-free diet based on chicken meat formula. At four months after total of 15 ml cow's milk she had an acute anaphylactic reaction. She recovered and is well on milk-free diet. Thus, this infant has recovered from necrotizing enterocolitis but has evidence of persistent cow's milk protein intolerance. The hypothesis is put forward that in this child cow's milk protein played a major role in the pathogenesis of necrotizing enterocolitis.


Characterization of major glycolipids in bovine erythrocyte membrane. Several neutral glycolipids and gangliosides were isolated from bovine erythrocyte stroma. Their structures were determined by partial acid hydrolysis, methylation analysis, periodate oxidation and CrO3 oxidation. Two major neutral glycolipids were characterized as lactosylceramide and galactosyl(alpha1--3)galactosyl(beta1--4)N-acetylglucosaminyl(beta1--3)galactosyl(beta1--4)glucosyl(beta1--1)ceramide. Two major gangliosides were N-glycolylneuraminosyl(2--3)galactosyl(beta1--4)glucosyl(beta1--1)ceramide and N-glycolylneuraminosyl(2--3)galactosyl(beta1--4)N-acetylglucosaminyl(beta1--3)galactosyl(beta1--4)glucosyl(beta1--1)ceramide. Minor glycolipids were glucosyl- and galactosylceramide, glucosamine-containing tri- and tetraglycosylceramide, glucosamine-containing disialosylhexaglycosylceramide, and gangliosides containing N-acetylneuraminic acid. The ceramide moiety of each glycolipid contained perdominantly d18:1 sphingosine, and normal fatty acids of C16:0, C22:0, C24:0, and C24:1.


Synthesis of a reactive ATP analog and its preliminary application as an affinity labeling reagent. A reactive ATP analog, N6-(6-bromoacetamidohexyl)-AMP.PCP, was synthesized in an attempt to covalently label the binding sites for adenine nucleotides, especially ATP, of various enzymes which utilize adenine nucleotides as substrates, cofactors, inhibitors or allosteric effectors. This reagent rapidly inactivated rabbit muscle glyceraldehyde 3-phosphate dehydrogenase (GPD), myokinase (MK), and creatine kinase (CK) under very mild conditions. Adenine nucleotide substrates prevented the inactivation. In the case of GPD, complete inactivation was observed when 1 mol of the reagent per mol of enzyme subunit was incorporated into the enzyme. These results indicate that the present ATP analog may be useful as an affinity labeling reagent for various adenine nucleotide-dependent enzymes.


Monomeric selectively S-alkylated derivatives of seminal ribonuclease: preparation and properties. Procedures are described for preparing monomeric selectively S-carboxamido-methylated and S-aminoethylated derivatives of seminal ribonuclease. The main properties of the derivatives, including their extinction coefficients, have been determined. Their catalytic activities and that of the S-carboxymethyl derivative have been tested. On double-stranded RNA as a substrate the monomeric derivatives are less active than the native dimeric enzyme, but much more active than pancreatic ribonuclease. On yeast RNA as a substrate the amino-ethyl derivative is found to be less active (80%) than the native enzyme, while the other two are over 30 percent more active. The monomers are stable in solution and when lyophilized from acetic acid solution do not associate to the same extent as pancreatic or native seminal ribonucleases.


Structural changes at the active site of pyruvate kinase during activation and catalysis. The distance between the obligatory monovalent and divalent cations at the active site of pyruvate kinase (rabbit muscle) has been used to monitor structural changes at various successive intermediates in the catalytic reaction and to relate structure and activity of substrate analogs. Determinations of distance were obtained from measurements of the longitudinal relaxation rate (1/T1) of the methyl protons of methyl ammonium ion, which was affected by the activating divalent cation Mn2+ in the various enzyme complexes. A frequency dependence of the 1/T1 effects with several complexes indicats that the observed relaxation rate changes are modulated by changes in the cation-cation distances. The results are interpreted as a sequence of structural changes at the active site which occur upon substrate binding, catalysis, and product release. Substrate analogs which are good analogs of phosphoenolpyruvate by kinetic criteria induce structural changes analogous to the substrate. An attempt is made to correlate the cation-cation distance changes to other structural changes reported for pyruvate kinase.


A portion of all major classes of histone messenger RNA in amphibian oocytes is polyadenylated. Ovaries of the amphibian Triturus viridescens contain a considerable amount of 7 to 12 S RNA which fractionates with poly(A)+ RNA on oligo(dT)-cellulose column chromatography and directs the synthesis of all five classes of histones in a wheat germ cell-free system. The polyadenylate tracts associated with this 7 to 12 S poly (A)+ RNA are heterogeneous in length, ranging from approximately 60 to 120 nucleotides. Partially purified subfractions of histone mRNA templates were isolated from this RNA by preparative polyacrylamide gel electrophoresis. The correlation between the poly(A) content, the template activity in vitro, and the cell-free products encoded by these RNA subfractions suggests that Triturus ovary RNA contains poly(A)+ histone mRNA. The 7 to 12 S poly(A)+ RNA, but not 7 to 12 S poly(A)- RNA, is an effective template for the avian myeloblastosis virus reverse transcriptase-directed synthesis of complementary DNA (cDNA) in the presence of oligo(dT) primer. In the absence of primer, virtually no cDNA is synthesized. When cDNA complementary to 7 to 12 S poly(A)+ RNA is hybridized in situ to Drosophila melanogaster polytene chromosome preparations. cDNA hybrids are found primarily in the region 39D-E, the locus of the histone genes in Drosophila. This cDNA also hybridizes to the sea urchin histone gene sequences contained in the chimeric bacterial plasmids pSp2 and pSp17. These results provide strong evidence for the reality of polyadenylated histone mRNA in Triturus ovary.


D-3-Phosphoglycerate dehydrogenase from chicken liver. I. Purification. A method is described for the preparation of homogeneous D-3-phosphoglycerate dehydrogenase from chicken liver in amounts sufficient for structural studies. The procedure utilizes ammonium sulfate precipitation, blue dextran-Sepharose chromatography, ion exchange chromatography on phosphocellulose, and crystallization. Previous reports of instability of the enzyme have been shown to be due to proteolysis in the crude extract which can be effectively prevented by leupeptin. The purified enzyme is a basic protein with a pI of 8.95 as measured by isoelectric focusing. The extinction coefficient at 278 nm of a 1% solution is 5.3.


D-3-Phosphoglycerate dehydrogenase from chicken liver. II. Chemical and physical properties. The molecular weight of chicken liver D-3-phosphglycerate dehydrogenase, as measured by sedimentation equilibrium analysis, was found to be 165,000, consistent with the measured sedimentation coefficient of 8.13 S. Sedimentation equilibrium studies in 6 M guanidine hydrochloride and gel electrophoresis in sodium dodecyl sulfate gave molecular weights of 40,000 to 43,000, indicating that the protein is composed of 4 subunits of equal molecular weight. Fractionation of the soluble tryptic peptides of the S-[14C]carboxymethylated enzyme by ion exchange chromatography and unidirectional paper electrophoresis indicated a maximum of 40 major peptides, which is approximately one-fourth of the number of tryptic peptides expected from the lysine plus arginine content of 157 of the tetramer. This observation suggests that the subunits possess identical or very similar amino acid sequences. The purified enzyme is a basic protein with an isoelectric point of 8.95. Amino acid analysis and titration with p-hydroxymercuribenzoate indicated that the 12 half-cystinyl residues present/subunit exist in the free sulfhydryl form. NH2-terminal analysis in an automatic protein sequenator indicated that the polypeptide chains of the protein are blocked.


Structural basis for the specific activation of human enteropeptidase. Human cationic trypsinogen is activated by human enteropeptidase much more readily than bovine trypsinogen, the ratios kcat/Km being 330 and 11 mM-1S-1, respectively. Conversely, porcine enteropeptidase activates bovine trypsinogen much more rapidly (kcat/Km = 630 mM-1S-1) than human cationic trypsinogen (kcat/Km = 2.4 mM-1S-1). The primary structure of the activation region of human cationic trypsinogen has been investigated in an attempt to elucidate the basis for these findings. The sequence of the first 12 residues at the NH2-terminus of human cationic trypsinogen has been shown to be Asp-Lys-Ile-Val-Gly-Gly-Tyr-Asn-Cys-Glu-Glu-Asn. Furthermore, the activation peptide derived from human cationic trypsinogen has been isolated and shown to be the dipeptide Asp-Lys. This result is in contrast to the Val-(Asp)4-Lys activation peptide from bovine trypsinogen and demonstrates that human cationic trypsinogen does not contain the (Asp)4 sequence present in many other mammalian trypsinogens. It is proposed that the high degree of specificity for activation of human cationic trypsinogen by human enteropeptidase is due to the preferential recognition of the novel activation peptide sequence in the human zymogen. Thus, these two functionally related proteins, cationic trypsinogen and enteropeptidase, may have evolved in a parallel manner in the human lineage.


The non-tubulin component of microtubule protein oligomers. Effect on self-association and hydrodynamic properties. We have investigated the association of non-tubulin microtubule proteins with tubulin to form the ring-shaped oligomeric structures found in microtubule preparations. We have found that the two oligomeric species present in our preparations of microtubule protein (s020,w = 18 and 30 S) each require non-tubulin factors for their formation. Two types of non-tubulin protein, the high molecular weight proteins (HMW) and the tau proteins were found to be active in ring formation. The HMW proteins promoted the formation of the 30 S oligomer, while the tau proteins promoted the formation of an oligomer of s020,w = 20 S. Analysis of the 30 S oligomer by gel filtration chromatography showed that the ratio of HMW proteins to tubulin was about twice that in the microtubule. The HMW proteins could be destroyed by exposure to trypsin, resulting in a marked increase in the sedimentation coefficient of the 30 S oligomer to 39 S. The 20, 30, and 39 S species were identified as rings by electron microscopy. The identity of the 18 S structure as a ring was called into question. Our data indicate that the 20 S species is a single ring and that the 30 S oligomer is a two-layered ring bearing HMW projections which contribute substantial hydrodynamic drag to the particle. We compare the organization of tubulin subunits and HMW molecules in the 30 S ring with the organization of these components in the microtubule and suggest that the organization in the ring is conserved in the microtubule.


Electron microscopy of metal-shadowed and negatively stained microtubule protein. Structure of the 30 S oligomer. Microtubule protein purified from porcine brain was fixed at low protein concentration with glutaraldehyde under conditions which maximize the relative concentration of the ring-shaped 30 S oligomer. Fixed oligomer was separated from glutaraldehyde and other protein species by column chromatography. The fixed, isolated oligomer was deposited on electron microscopy grids, dehydrated, and then critical point-dried before shadow-coating with carbon/platinum alloy at a fixed angle. Analysis of the shadow lengths observed by electron microscopy revealed that the height of the 30 S oligomer is 15 nm. Microtubule protein deposited on electron microscope grids at high protein concentrations was examined by the negative stain technique and found to contain apparent stacks of oligomer from which the number of tubulin dimers per turn of the ring and the distance between turns could be determined. The number of subunits per turn was determined as 13.8. The distance between turns was found to be 7.4 nm, indicating that the 15 nm high, shadowed oligomers consisted of two turns. Additional information from the literature is considered and a model is presented for the oligomer. The model is a helix of 29 tubulin dimers and five high molecular weight protein molecules arranged so as to preserve intersubunit bonding patterns found in microtubules.


Sedimentation velocity analyses of the effect of hydrostatic pressure on the 30 S microtubule protein oligomer. Increasing hydrostatic pressure in the analytical ultracentrifuge by increasing rotor velocity and overlayering protein samples with oil caused a depolymerization of the 30 S oligomer of microtubule protein. This results indicates that the reaction of 6 S microtubule protein to form the oligomer was accompanied by a positive volume change. The effect of hydrostatic pressure on the 6 S to 30 S transition was employed to demonstrate the presence of a rapidly reversible equilibrium between these components by showing polymerization or depolymerization of the oligomer during the course of ultracentrifugation. The magnitude of the partial specific volume change accompanying this reaction was estimated from mass fraction measurements of microtubule protein solutions at a variety of hydrostatic pressures to be about 9 X 10(-4) ml g-1.


Skeletal muscle acetylcholine receptor. Purification, characterization, and turnover in muscle cell cultures. Acetylcholine receptor has been purified from embryonic skeletal muscle cells grown and allowed to differentiate in tissue culture. The polypeptide composition of purified receptor has been determined by two-dimensional electrophoresis. The purest preparations are composed of a single Mr = 41,000 class of polypeptide which exhibits some charge heterogeneity. By high resolution two-dimensional electrophoresis a spot corresponding to acetylcholine receptor was localized among total proteins of muscle membrane extracts. Synthesis of this component is shown to be developmentally regulated. Quantitative analysis of receptor synthesis and degradation has led to the conclusion that receptor is one of a class of proteins whose synthesis is tightly regulated during terminal steps of myogenesis.


Cytoplasmic filaments and gap junctions in epithelial cells and myofibroblasts during wound healing. During the healing of an experimental skin wound, epidermal cells and granulation tissue fibroblasts (myofibroblasts) develop an extensive cytoplasmic contactile apparatus. Concurrently, the proportion of epidermal cell surface occupied by gap junctions increases when compared to normal skin, and newly formed gap junctions appear between myofibroblasts; this suggests that epidermal cell migration and granulation tissue contraction are synchronized phenomena.


The spermatozoon of arthropoda. XXX. The multiflagellate spermatozoon in the termite Mastotermes darwiniensis. In this paper the spermatozoon of the termite Mastotermes darwiniensis is described. It is the first example of a multiflagellate sperm cell in animals. The sperm consists of a conical head and 100 flagella. Other remarkable features of this sperm cell are the absence of an acrosome, the presence of centrioles containing doublet microtubules instead of triplets, and the presence of axonemes devoid of central tubules and with doublets bearing only one arm. The flagella are feebly motile.


Fluorescein colchicine. Synthesis, purification, and biological activity. The synthesis of a fluorescent colchicine derivative permits the localization of colchicine-binding receptors in cells. Fluorescein colchicine (FC) was prepared by the addition of fluorescein isothiocyanate to deacetyl colchicine. The product, FC, was separated from the reactants by thin-layer chromatography (TLC). The purity of FC was demonstrated by TLC, UV spectral analysis, and analysis of the kinetics of photodecomposition. FC inhibited [3H] colchicine binding to purified brain tubulin. The biological activity of FC was compared to the activity of unlabeled colchicine on mitosis, motility, secretion, and myogenesis. The effects of FC were identical to those of unlabeled colchicine in all biological systems tested. The results demonstrate that FC may be substituted for colchicine in biological experiments without significant loss in specificity or effectiveness.


Rat liver nuclear skeleton and small molecular weight RNA species. Small molecular weight RNA species (smwRNAs) were studied in rat liver nuclei with and without chromatin as well as with and without nuclear envelope and nucleoplasm. From all the species identified, only two, N5 and 5Sb, were related to ribosomes. The others were localized exclusively in the nuclear skeleton or the spongelike network that was described in the preceding communication. This network or protein matrix contains a less abundant but exclusive set of molecules designated 5Sa, N1, and 4.5S, as well as other more abundant molecules which also exist in rat liver endoplasmic reticulum but not in polysomes or postribosomal RNP complexes. The smwRNAs behave like HnRNA; they remain located in the nuclear skeleton when nuclei are deprived of nucleoplasm and chromatin. With the information presently available, it is not possible to know whetherer both species are in the same or different RNP complexes and whether some of the smwRNAs contribute to the architecture of the nuclear skeleton. Distinct from any other nuclear RNA species, smwRNAs have two unique properties: facility of extraction, and resistance to nuclear ribonuclease digestion.


Meiosis in Coprinus. VIII. A time-course study of the fusion and division of the spindle pole body during meiosis. The time-course study of meiosis in the fungus Coprinus cinereus (C. lagopus) by electron microscopy reveals that two monoglobular spindle pole bodies (SPB's) of prekaryogamy nuclei come together during karyogamy and are fused. The fusion SPB of postkaryogamy nucleus persists through zygotene and pachytene as evidenced by the presence of axial components and synaptonemal complexes. At early diplotene, the SPB divides. The divided SPB takes on a diglobular form, which grows in size to form two daughter SPB's. These separate and move to opposite poles at metaphase I.


Radioimmunoprecipitation assay for quantitation of serum antibody to the hemagglutinin of type A influenza virus. A double-antibody radioimmunoprecipitation (RIP) assay has been developed to provide a sensitive and specific measure of antibody to hemagglutinins of H3N2 influenza viruses. Chloramine T was used to radiolabel purified hemagglutinins to high specific activity without loss of antigenicity. The purity of the labeled hemagglutinin was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, which also established that both the HA(1) and HA(2) polypeptides were iodinated. Radiolabeled hemagglutinins with a specific activity that did not exceed 12 muCi/mug of protein could be maintained for up to 30 days at -70 degrees C in the presence of supplemental protein. The RIP assay was compared with conventional methods, hemagglutination inhibition and viral neutralization tests, using H3N equine 1 hybrid viruses for determining serum antihemagglutinin antibody titers. The geometric mean titers for human convalescent sera after infection with A/England/72 virus were 118, 161, and 18,822 for hemagglutination inhibition, viral neutralization, and RIP tests, respectively, and the three tests demonstrated significant rises in antihemagglutinin antibody titers with equal efficiency. In general, a positive correlation existed between antihemagglutinin antibody titers determined by these three procedures; however, the antibody level determined by RIP assay for each individual could not be related to hemagglutination inhibition or viral neutralization titers by a constant factor. A similar lack of a constant relationship was found by using hyperimmune guinea pig antisera, which suggests that the RIP assay can detect antibody populations that exhibit differing efficiencies for inhibition of viral hemagglutination and replication.


[The testis in a 46, XX male. Apropos of an ultrastructural examination]. The ultrastructure of a testicular biopsy was carried out in a man with 46 XX chromosomes who had a masculine phenotype and in whom the hormone levels showed that the interstitial tissue of the gland was functioning well. Observations showed that the exocrine appearance of the testis was comparable to that described in Klinefelter's syndrome, namely germinal aplasia with invasion by Sertoli cells. On the other hand the interstitial glandular tissue showed morphological characteristics associated with normal androgen activity, which is unusual to find in Klinefelter's syndrome. Lipid inclusions, pigmentation and Reinke cristalloids were found. The cytogenetic techniques that happened to be used did not manage to show up even a small fragment of the Y chromosome. Various theories have been considered to explain the constitution of such a phenotype with this chromosome abnormality.


[Fetal hypothyroidism: a complication of amniofetography. Apropos of 3 cases]. The authors present three cases of fetal hypothyroidism following amniofetography, after they have reviewed the technique and complications of this procedure. (For outlining the fetus by injecting radio-opaque iodides into the amniotic sac). These are cases of hypothyroidism in the fetus caused by overdosage with iodides. The fetal thyroid is blocked by the large quantity of iodine which is necessary for amniofetography. The indications then for amniofetography should be reviewed. The only valid indication that remains is for the diagnosis of neurological malformations and especially spina bifida. Whenever amniofetography has been carried out the newborn should be assessed for thyroid function routinely.


[Immunologic and endocrinologic considerations apropos of a case of acardia]. The deficient development of the peripheral immunopoietic system in acardiac acephalous monsters cannot be explained only by the absence of the thymus. Similarly the adrenal gland hypoplasia does not depend upon the absence of the pituitary gland due to the concurrent condition of acephaly. The author suggests that the failed development of the above mentioned structures depends respectively upon disorders in the peripheral "microenvironment" which is needed by immature lymphoid cells to establish themselves in the peripheral lymphoid structures and upon the decreased sensibility of the adrenal glands to the corticotrophic stimulus. A hypothesis about the frequency of the association between acephaly and acardy is proposed. Finally the author proposes a study program in such cases.


Metabolism of androstenedione in skin and serum levels of gonadotrophins and androgens in prepubertal boys with hypospadias. The metabolism of 4-[1,2-3H]androstene-3,17-dione in the prepuce, axillary skin and skin from the arm was investigated in 27 boys operated for phimosis (controls) and 13 unselected boys with hypospadias (a congenital defect of the male urethra). In all types of skin investigated, androstenedione was metabolized to 5alpha-androstane-3,17-dione, 3alpha-hydroxy-5alpha-androstan-17-one, 3beta-hydroxy-5alpha-androstan-17-one and testosterone. Conversion to testosterone was found in the prepuce of two out of 11 boys with hypospadias. Mild forms of hypospadias in the age group 1--4 years had a higher level of 5alpha-reductase activity in the prepuce than controls in the same age group (P less than 0.05); no such differences were found in the few severe cases of hypospadias in this group. No other differences in 5alpha-reductase activity were found between hypospadic boys and controls. The ratio of 5alpha-reductase activity in the prepuce: 5alpha-reductase activity in skin from the arm was significantly higher (P less than 0.05) in hypospadic boys than in controls in the age group 1--4 years. Serum levels of LH and FSH were the same in normal and hypospadic boys but the concentration of prolactin in the serum was lower in boys with hypospadias compared with control subjects in the age group 1--4 years (P less than 0.005). No differences were found in serum concentrations of androstenedione, testosterone, oestradiol and testosterone-binding globulin.


Correlation between the effects of neuroleptics on prolactin release, mammary stimulation and the vaginal cycle in rats. The correlation between hyperprolactinaemia induced by the administration of neuroleptic drugs, disturbances of the vaginal cycle and mammary gland stimulation in rats was investigated as a test model simulating the clinical syndrome of hyperprolactinaemia and amenorrhoea with anovulation. In acute experiments in which clozapine, sulpiride and chlorpromazine were administered orally to rats of both sexes, there were rapid increases in the level of prolactin in the serum with peak values between 15 and 60 min. The responses of female rats to various doses of sulpiride were consistently higher than those of male rats. Hyperprolactinaemia induced by sulpiride in dioestrous rats failed to desensitize the ovaries to the ovulatory effect of exogenous luteinizing hormone releasing hormone. Studies of these substances and of metoclopramide, haloperidol and thioridazine were then carried out in females rats by daily oral administration over a period of 13 days. The increases in the level of prolactin in the serum were paralleled by disruption of the vaginal cycle up to and including constant dioestrus and by mammary gland stimulation which, like the preceding phenomena, showed dose-dependence. The potencies of these six neuroleptics, as estimated from their effects on the mammary gland, appeared to be haloperidol greater than sulpiride greater than or equal to metoclopramide = thioridazine greater than chlorpromazine greater than clozapine.