The effect of cobaltous chloride on liver haem metabolism in the rat. Evidence for inhibition of haem synthesis and for increased haem degradation. After a single dose of cobaltous chloride there was a marked inhibition of liver 5-aminolaevulinate (5-ALA) synthetase (at 1 h) and this was followed in turn by a stimulation of haem oxygenase (at 3 h) and by a return of the synthetase activity to normal or above normal (at 17 h). Bile cannulation experiments were performed 1 and 17 h after administration of CoCl2. At 1 h there was a marked decrease in bile porphyrin content, no change in bile concentration of bilirubin, but a decrease in the conversion of [14C]-5-ALA to bilirubin and to liver haem. At 17 h, an the other hand, the bile excretion of both porphyrins and bilirubin was significantly greater than in controls and more radioactivity (from [14C]-5-ALA) appeared in the bile as bilirubin. It is concluded that the effects of cobalt on liver haem metabolism are complex and time-dependent. There is first inhibition of liver haem synthesis at two different steps of the pathway (synthesis of 5-ALA and conversion of 5-ALA to haem), with diversion of [14C]-5-ALA into a relatively stable liver pool different from haem; and at a later stage there is also an increase in the rate of liver haem degradation.


Multicompartment analysis of 14C-labelled coproporphyrin and uroporphyrin kinetics in human beings. 14C-labelled coproporphyrin I and III and uroporphyrin I were injected intravenously into healthy human subjects. Two experiments were performed with each porphyrin. Counting of blood, urine and faeces was made as a function of time. The coproporphyrin data were simulated by means of an analogy computer by using a four-compartment model with a delay pool. The physiological counterparts to the compartments were considered to be liver, blood and gut. From the blood radioactivity disappearance curve of uroporphyrin three exponential components could be worked out. The least square method was used for the solution of the plasma and urine radioactivity curves. A three-compartment model was constructed to illustrate the kinetics of uroporphyrin I. Coproporphyrin I and III behaved essentially similarly, whilst the kinetic behaviour of uroporphyrin I was completely different from that of coproporphyrins. A rapid hepatic uptake, preferential faecal excretion and a significant enterohepatic circulation were features of coproporphyrin kinetics. Over 70 per cent of the tracer activity of uroporphyrin I was found to accumulate to a rather large pool (volume 16--28 times the plasma volume) from where the back flow to plasma was rather slow. Practically all the uroporphyrin was excreted via urine.


The enzymatic conversion of heme to bilirubin in vivo. The conversion of hemoglobin to bilirubin IXalpha in vivo probably proceeds via alpha-hydroxyheme to carbon monoxide and biliverdin-iron complex; after hydrolysis, biliverdin IXalpha is liberated from the iron complex and reduced enzymatically to bilirubin IXalpha. For these reactions molecular oxygen, NADPH and a functioning microsomal electron transport system are needed to generate a reaction radical to transform the heme at the alpha-bridge to alpha-hydroxyheme. After that the reaction proceeds spontaneously. During this reaction sequence heme is readily bound to an insoluble and substrate-inducible protein, which behaves as a stereospecific oxidase. The crucial point in all likelihood is the nature of this heme-binding protein. It seems to be a large protein molecule having a molecular weight in excess of 200,000 and located within the membrane of the endoplasmic reticulum. The conversion of heme to bilirubin has thus some characteristics of nonenzymatic coupled oxidation and, on the other hand, of mixed-function oxidations catalyzed by cytochrome P-450. The exact nature and the regulation of this process, however, appears to be quite different from that manifested with drugs and steroids.


Turnover of erythrocyte protoporphyrin, with special reference to bovine porphyria and iron deficiency anemia. Available data from human subjects suggests that most of the pools of free protoporphyrin in circulating erythrocytes have T 1/2 values which range from less than 1 hour to approximately 2 weeks. An exception was observed in 2 cows with erythropoietic ("congenital") porphyria. During the several months which followed the simultaneous injection of 14C- and 3H-labelled glycine and ALA, changes in the specific activities of both free- and hemoglobin-protoporphyrin were similar. Since erythropoietic porphyria in the bovine differs from that in the human mainly in the elevated values of free erythrocyte protoporphyrin in the former, it is suggested that this prolonged T 1/2 (equal approximately to the red cell life span) may account for the elevated values seen in the bovine. Variable ratios of red cell/fecal protoporphyrin concentrations reported in other diseases are also believed to be due largely to T 1/2 differences in porphyrin pools. Recorded fluorescence and excitation spectra of blood showed the single fluorescence band of "free" porphyrin in bloods of normal and porphyric cows and of patients with erythropoietic porphyria, protoporphyria, and sideroblastic anemia. The 2 band zinc complex spectrum was found, as expected, in blood of subjects with Pb poisoning and Fe deficiency anemia. T 1/2 values of 1 to 2 weeks were calculated for red cell protoporphyrin from previously published data of patients with Fe deficiency anemia who were treated effectively with iron.


Special clinical, light and electron microscopic aspects of acute intermittent porphyria. Clinical symptoms in 26 patients with acute intermittent porphyria (AIP) were correlated with light and electron microscopic findings. Abdominal pain, neuropsychiatric disorders and pathologic urine tests were predominant. Light and electron microscopic pictures of the central nervous system, the liver and the kidney were shown.


The binding and transport of heme by hemopexin. Hemopexin binds proto-, meso-, or deutero-ferriheme with high affinity, forming an equimolar, low-spin complex. The ferroheme-hemopexin complex, which coordinates with CO and readily autoxidizes, is also low-spin. Formation of the ferriheme-hemopexin complex requires essential histidine and tryptophan residues and induces changes in the protein's tertiary structure. These changes may be important for the uptake of the heme-hemopexin complex by hepatocytes. Hemopexin also binds other porphyrins including protoporphyrin IX, and uro- and coproporphyrins I and III in a 1:1 molar ratio, but they are readily displaced by heme and do not produce discernable changes in the protein's conformation. In preliminary experiments, a selective interaction in vitro between heme-hemopexin and isolated rat hepatocytes has been demonstrated. This information is used as the basis for proposed models of the heme-binding site of hemopexin and of the interaction of heme-hemopexin with the parenchymal cells of the liver.


Catabolism of homologous and heterologous hemopexin in the rat and uptake of hemopexin by isolated perfused rat liver. Iodinated hemopexin (Hx) from three different species, rabbit, human and rat, was injected into rats and its clearance from the plasma measured. Rabbit, human and rat apo-Hx were cleared from the plasma with a T 1/2 of 20--31 h, 31--32 h, and 48--60 h respectively. Heme injection (10 mg/kg) after equilibration of the protein immediately accelerates elimination of the hemopexins of all three species (T 1/2 of 4.5 to 10 h). This indicates that the two heterologous hemopexins maintain their function in heme transport. The T 1/2 of rabbit and human Hx return to pre-heme injection values 16 to 20 hours after the injection of heme. For rat Hx, however, the T 1/2, which was 54 +/- 3.5 h before heme injection, was reduced to 25 +/- 1.3 h 20 hours after heme injection. Administration of 1.2--1.3 mg of protoporphyrin IX or uroporphyrin III, after equilibration of iodinated rat Hx, did not change the T 1/2 of the protein, whereas the same amount of coproporphyrin III significantly reduced its T 1/2 from 54 +/- 3.5 to 37 +/- 0.4 h. In addition, the uptake of rat apo-Hx, heme-Hx and albumin by rat liver tissue was measured in an isolated liver perfusion system using radioiodinated proteins screened in vivo. The uptake of apo-Hx by the liver after 2 h (46.8 ml/100 g) was less than that of heme-Hx (67.3 ml/100 g). The amount of apo-Hx and heme-Hx associated with the liver, relative to that circulating in the perfusate, was greater than that of albumin (12.1 ml/100 g). These results are considered to represent selective uptake of Hx by the liver induced by its interaction with heme.


Isolation and partial characterization of rat liver porphyrin binding proteins. 14C-coproporphyrin III was added to hemoglobin-free cytoplasmic rat liver extract. Molecular sieving was performed on Sephadex G-75 column. The protein content of each fraction was measured at 280 nm in double beam spectrophotometer and the radioactivity by means of liquid scintillation counter. In several runs on Sephadex we found four main radioactive peaks which correlate to four porphyrin binding proteins in rat liver cytoplasm. One fraction, which probably is identical with Y protein or ligandin, was further purified and characterized by means of different chromatographic techniques.


The transport of hemin and protoporphyrin across the plasma membrane of chick embryo liver cells in culture. Primary monolayer cultures of chick embryo hepatocytes can be cultured in a chemically defined medium (Ham F-12) containing insulin. The absence of serum from the medium permitted a study of the effects of added serum proteins on the transport of hemin and protoporphyrin across the plasma membrane of the hepatocyte. As the criterion of hemin uptake we used its unique and selective activity in repressing the induced synthesis of delta-aminolevulinate synthetase by various chemicals. Movement of hemin into the cells is rapid and does not require added serum proteins. Hemin represses the induced synthesis. The repression by hemin is decreased 50% when the molar ratio of hemin to human serum albumin (6.5 muM) is 1 :2, i.e., where the calculated concentration of dissociated hemin is 10(-8) M. Apparently serum albumin does not enter the cells; it decreases entry of hemin into the cells by virtue of its high affinity for hemin. Compared to human serum albumin, bovine serum albumin and chicken serum albumin, under the same conditions, have a much lower affinity for hemin and scarcely influence the repression effect by hemin. Protoporphyrin can be specifically caused to accumulate in the cytosol, and uroporphyrin in the nucleus of the hepatocytes by the use of different inducers of delta-aminolevulinate synthetase. Protoporphyrin, but not uroporphyrin, is released rapidly from the cells when the moles of human serum albumin added to the medium are 5 times that of porphyrin. This culture system may provide a useful model for studying the mechanism of transport of organic anions across the hepatocyte plasma membrane.


The induction of heme oxidation in various tissues by trace metals: evidence for the catabolism of endogenous heme by hepatic heme oxygenase. Cobalt is a potent inducer of hepatic heme oxygenase and concomitantly decreases microsomal cytochrome P-450 content in liver cells. Studies in which microsomal heme was labelled with 14C-beta-aminolevulinic acid showed that the decline observed in P-450 follows the induction of heme oxygenase indicating that endogenous heme is a substrate for this enzyme. The inducing effect of cobalt on heme oxygenase in liver extended to other organs such as heart, renal cortex and medulla, lungs and intestinal mucosa but not to the spleen and brain. In kidney and heart, cobalt resulted in decreases in microsomal and mitochondrial contents of hemoproteins. In addition ALAS activity of kidney was depressed by the metal. Other metals which chelate with sulfhydryl groups (e.g., Cr, Fe, Ni, Cu, Zn, Cd, Hg, Pb) were also shown to induce heart and renal heme oxygenase activities. Covalent binding of cobalt with mercaptans inactivated the metal with respect to its induction of heme oxygenase and degradation of hemoproteins.


Hepta- and hexa-carboxylic porphyrinogen intermediates in haem biosynthesis. In the course of our studies on intermediates in normal and abnormal metabolism of porphyrins we have synthesised a number of porphyrins related to uroporphyrin-III and compared them with materials isolated from natural sources. In the present paper we show that the corresponding porphyrinogens are all metabolised to protoporphyrin-IX by haemolysates of chicken erythrocytes, but at different rates. The results are discussed in relation to our conclusions concerning the preferred pathway of degradation of uro'gen-III to coproporphyrinogen-III, which indicate a clockwise sequence of decarboxylation reactions.


The biosynthesis of delta-aminolevulinic acid from the intact carbon skeleton of glutamic acid in greening barley. The formation of delta-aminolevulinic acid in mammals birds, yeast and some bacteria is known to take place by the ALA-synthetase assisted coupling of glycine with succinylCoA. Plants, however, form the bulk of their delta-aminolevulinate in another way. We present evidence here that the intact, 5-carbon chain of glutamate becomes that of delta-aminolevulinate. Greening barley was fed specifically labelled glutamate and levulinate to create a pool of labelled delta-aminolevulinate. Levulinate, a competitive inhibitor of ALA-dehydratase, prevents the metabolism of delta-aminolevulinate. The carbon chain of the labelled delta-aminolevulinate was broken into formaldehyde and succinate by periodate to determine the position of the label. It was found that the C1, carboxyl carbon of glutamate becomes the amino-bearing (C5) carbon of delta-aminolevulinate and forms the formaldehyde on cleavage. delta-Aminolevulinate formed from C3,4-labelled glutamate bears its label in the succinate cleaved fragment. We conclude that during the light induced development of the plastid in barley the carbon chain of delta-aminolevulinate is formed from the intact chain of glutamate. ALA-synthetase catalyzed the formation of delta-aminolevulinate from glycine and succinylCoA cannot play a quantitively important role in the formation of chlorophyll.


Studies on the conversion of the cytosol delta-aminolevulinate synthetase to the mitochondrial enzyme. DELTA-Aminolevulinate (ALA) synthetase was induced in the rat by allylisopropylacetamide, and ALA synthetase in the liver cytosol fraction was partially purified. The cytosol ALA synthetase shows a molecular weight (M.W.) of about 600,000 by gel filtration and about 178,000 by sucrose density gradient centrifugation, whereas the mitochondrial enzyme gives M.W. about 110,000 by either method. In the presence of 0.25 M NaCl, however, the cytosol ALA synthetase showed M.W. 300,000--350,000 by gel filtration and gave M.W. 110,000 or a S value of 6.4 by centrifugation. Treatment of the cytosol ALA synthetase with papain or snake venom also yielded an entity which showed M.W. 110,000 by gel filtration as well as by sucrose density gradient centrifugation. When the 6.4 S enzyme once separated by centrifugation was combined with a fraction exhibiting about 4 S on centrifugation and was subsequently subjected to gel filtration, the ALA synthetase activity was recovered in the fraction corresponding to M.W. about 170,000. When the combination of the 6.4 S enzyme and a fraction of about 8 S was examined by gel filtration, the ALA synthetase activity was eluted showing M.W. about 250,000. The ALA synthetase in the cytosol fraction seems to exist as a complex composed of an ALA synthetase of M.W. 110,000 and a 4 S protein and an 8 S protein to form a larger size of the enzyme (M.W. 300,000) which further aggregates to give M.W. about 600,000. Physiologically, the complex may be subjected to a limited proteolysis so as to lose the capacity to form complex and aggregates, followed by incorporation of smaller sizes of the enzyme into the mitochondria.


Pentacarboxylic intermediates in haem biosynthesis. The incorporation of the four possible type III porphyrinogens into protoporphyrin is described and their various rates or conversion are discussed. The techniques used involved the incubation of the porphyrinogens in chicken red cell haemolysates followed by extraction and characterisation of the end products on HPLC and TLC. Porphyrin 5 b c d was shown to be more slowly incorporated than porphyrins 5 a b d, 5 a c d and 5 a b c. The relevance of these findings to an understanding of the porphyrin excretion pattern in hepato-erythrocitic porphyria is considered.


Porphyrin biosynthesis from ALA and PBG by human erythrocytes in porphyrin disorders. Kinetic studies of the isomer series I and III. Porphyrin biosynthesis from exogenous ALA and PBG was observed over a period of from 1 to 8 hours in hemolysates of red blood cells from patients with AIP, PCT, and EPP. In all these conditions all porphyrins in the biosynthetic pathway through to PROTO were formed, with URO as the dominant component. Compared to normals, total porphyrin synthesis was lowered during the entire incubation period in both AIP and EPP, but enhanced in PCT. In hemolysates after 1 hour the second component of the pattern in AIP is COPRO, in PCT HEPTA and in EPP PROTO. Of the isomers, COPRO III predominates in all conditions (greater than 80%), whereas URO contains more isomer I than III. In intact red cells from controls, AIP, and PCT, COPRO is always the main component formed from ALA. Total synthesis is diminished in AIP by about 50%, in analogy to the findings on hemolysates from AIP. In heated hemolysates URO formation is diminished by half in AIP, but nearly double control values in PTC. In this sense URO'gen-I-synthase activity is increased in PCT. These findings reflect the underlying enzymatic defect: Diminished URO'gen-I-synthase activity in AIP, diminished URO'gen decarboxylase activity in PCT, and diminished activity of ferrochelatase in EPP.


[Glucocorticoids receptors in rat testis: their role in Leydig cells specific function and DNA synthesis (author's transl)]. Rat testicular cytosol binds specificity dexamethasone (KD = 5 X 10(=9)M). After incubation of testicular slices with 3H-dexamethasone, at 30 degrees C for 2 hours most of the intracellular radioactivity is found inthe nuclei. Administration of h CG at pharmacological doses for three days to male rat during puberty increases plasma testosterone levels and 3H-thymidine incorporation into testicular DNA. Simultaneous administration of dexamethasone partially blocks both effects of hCG. These results suggest that interaction of glucocorticoïds with specific receptors controls both the specific function and the cells multiplication of rat Leydig cells.


[Quaternary structure of thyroglobulin (author's transl)]. Porcine and ovine thyroglobulins were purified from fresh glands by slice extraciton, precipitation with buffered ammonium sulfate and Sepharose 6B gel filtration using 0.1 M sodium phosphate buffer pH 7.2 at all steps. In contrast to thyroglobulin obtained from frozen glands, the proteins purified from unfrozen glands only showed the 19 S and 12 S species by electrophoresis in sodium dodecylsulfate polyacrylamide gels. After full reduction and S-alkylation, no qualitative changes were observed in e gel electrophoresis patterns as compared to the unmodified proteins. Species of apparent molecular weight corresponding to the native 12 S subunit was the major component strongly suggesting a mol. wt. of about 330 000 for the elementary peptide chains of pig and sheep thyroglobulin. This was confirmed by sedimentation equilibrium analyses in 6 M guanidinium. HCl, 0.1 M sodium phosphate pH 7.2 OF FULLY REDUCED AND S-alkylated thyroglobulins which showed a weight average molecular weight of 310 000 daltons.


[Hormonal control of prostaglandins biosynthesis during lipolysis in rat adipose tissue (author's transl)]. Lipolysis in rat fat pads was studied by determination of free fatty acid and glycerol production in various experimental conditions (absence or presence of glucose, adrenaline and insulin). These results were compared to the accumulation of endogenous prostaglandins E2 and F2 alpha during lipolysis. In the absence of glucose the prostaglandin production followed the adrenaline-ininduced fluctuations in released free fatty acids both in the presence or absence of insulin. In the presence of glucose and insulin, a drop in prostaglandin accumulation was observed whereas free fatty acid production was strongly stimulated. These results suggest either that free fatty acid composition is modified influencing the acitvity of prostaglandin synthetase, or that there exists a specific mechanism controlling prostaglandin synthesis.


[Comparison of the effect of TSH and fluoride on the adenylate cyclase activity of cold thyroid nodules (author's transl)]. Using a simple and reliable assay technique in which the actuel amount of cyclic AMP generated during incubation is determined by competitive protein binding assay, adenylate cyclase activity was measured in 7 cold nodules and 3 samples of normal thyroid tissue. Mitochondrial fraction prepared from surgically obtained tissues was used as the source of the enzyme. We observed an increased adenylate cyclase activity in cold nodules as compared to normal. More importantly, this activity was much more responsive to TSH in cold nodules than in normal (maximal stimulation: 1050+/-200% vs 320+/-120%). Also, maximal TSH stimulation was similar to that elucited by fluoride in the cold nodules but only one-third in normal tissue. These results suggest that higher efficiency of TSH in cold nodules may be related to increased TSH binding or coupling of TSH receptors with the caralytic subunit of adenylate cyclase.


[Hyperthyroidism by autonomous metastasis of thyroid carcinoma (author's transl)]. Nine years after surgical ablation of a trabeculo-vesicular carcinoma of the tyroid, a patient developped bone and liver metastasis. She had clinical signs of thyrotoxicosis, clear increase of blood T3 and sligh increase of T4. The TSH secretion was blocked. Exogenous TSH increased iodine uptake in the thyroid and not in the metastasis. After 2 doses of 120 mCi of 131I, she became hypothyroid, the liver was normal and the scan revealed the disappearance of uptake in thyroid and in metastasis. Such a clinical course was previously found in only 10 cases despite the frequent funcitonal differenciation of the metastasis of thyroid carcinomas.


[Acquired pseudohypoparathyroidism/(PHP) in adult coeliac disease (MCA)]. One case of chronic hypocalcemia and hyperphosphatemia, with an adult coeliac disease, hight P.T.H. level, very low rate of cyclic A.M.P. in urine, and normal rate of cyclid A.M.P. in plasma is described. It is the question of the pseudohypoparathyroidism acquirec as a result of an adult coeliac disease, because of after seven months of diet without gluten the clinical and biological improvement was observed with the increase of the cyclic-AMP clearance into the normal level and its great reactivity on the administration of exogenous PTH. We think to bring an additional analysis of the trouble of renal receptivity to parathyroid hormone.


[The Waardenburg-Klein syndrome].

[The Waardenburg-Klein syndrome]. A case of Waardenburg-Klein syndrome associated with a Hodgkin's disease is reported in a 29 year old female. Such an association seems to be fortuitous. The authors discuss the dermatological aspects of this rare disease, particularly the disturbances of cutaneous pigmentation. Ultrastructural study of depigmented skin indicates the melanocytes are absent and precise the place of this depigmentation in the group of genetic abnormalities of skin pigmentation.


Churg and Strauss granulomatous angeitis. The authors report a new case of Churg and Strauss allergic angeitis, observed during the patients lifetime. Severe asthma preceded the cutaneous lesions by 16 months. There was an eosinophilia of 66 p. 100 of 21,000 white cells. The histological picture was characteristic. A general review of the literature is made in the light of this case.


[Oestradiol receptors in foetal compartment (author's transl)]. Oestradiol concentration in the foetal plasma of guinea pig is 160-200 pg/ml (20-35 days of gestation), 200-300 pg/ml (35-45 days of gestation) and 170-450 pg/ml (45-60 days of gestation). After 3H-oestradiol administration to the foetus, this hormone circulates mainly as non-transformed 3H-oestradiol, 3H-oestrone and 3H-oestrone sulphate. 97-99% of the circulating oestradiol is found in unbound form. In vivo and in vitro studies in the foetus of guinea pig (40-55 days of gestation) show the presence of specific oestradiol receptors in the kidney, brain and Mullerian ducts of the foetus. The affinity constant in the cytosol of the brain is Kd=2 X 10(=9)M; in the cytosol kidney this value is Kd = 2.3 X 10(=10)M. A direct binding of 3H-oestradiol to the chromatin of the kidney was demonstrated with a Kd = 3.3 X 10(=10)M In the cytosol of Mullerian ducts the presence of a specific 3H-oestradiol macromolecule component with a sedimentation coefficient (in sucrose gradient) of 8-8.5S was detected. It is concluded that specific oestradiol receptors are present during foetal development in tissues such as kidney, brain and Mullerian ducts, but not in the liver.


[Eczema herpeticum. Clinical observation and ultrastructural study]. A child afflicted with atopic dermatitis developed a Kaposi-Juliusberg's syndrome. Virological and ultrastructural investigations proved a widespread herpes virus infection. Maturation of the virus within the nuclei of epidermal cells and its release into the cytoplasm and the intercellular space have been studied at the ultrastructural level and some peculiar data brought up.


[Effect of gamma irradiation on the production and degradation system of the second hormonal messenger (author's transl)]. Gamma irradiation of rat liver plasma membranes leads to a decrease of the adenylcyclase activities stimulated by glucagon and fluoride. The observed inhibition is more important for the activity stimulated with glucagon. The 5'-nucleotisade activity is not changed by irradiation. When the phosphodiesterase system is submitted to gamma irradiation, the radiosensibility of enzymatic complex is more important.


[Fibroepithelial tumors of Pinkus with extra-dorsal, lumbar and sacral localizations -31 cases]. In our series of 70 fibroepithelial  tumors of Pinkus, 31 tumors were located outside of the dorsal, lumbar and sacral areas. Moreover, we have noted in the literature 32 ectopic localizations. The clinical and evolutive characteristics are the same as for the dorsal, lumbar and sacral tumors. Thus, the extra-dorsal, lumbar and sacral tumors are much more frequent than usually believed; the clinical diagnosis is however much more difficult since one is deprived of the very important notion of lumbo-sacral localization.


[Regulation of chick oviduct DNA polymerases by steroid hormones (author's transl)]. DNA polymerase activity was measured in vitro in soluble cytoplasmic and nuclear fractions of chick oviduct, after in vivo hormone stimulation. By twelve hours after a subcutaneous injection of oestradiol, cytosolic and nuclear DNA polymerase activity had increased 2 to 5 fold over control levels ane continued to increase until 24 hours after the injection. Progesterone induced a slight rise in DNA polymerase activity and dihydrotestosterone had no effect. When oestradiol + progesterone were given together, the stimulating effect of oestradiol was cancelled for approximately 14 hours, contrasting with their synergist-c effect on transcription in this system. Various quantities and various sequences of hormone administration were studied, and some of the physico chemical parameters of the enzymes were determined after the different hormone stimulations. The demonstration of a sex steriod dependent regulation of chick oviduct DNA polymerases and the opposite effects of the oestradiol + progesterone combination on replication and on transcription can represent a new approach in the study of the mechanism of action of steroid sex hormones.


[Decreased number of insulin receptors in obesity: studies in the obese hyperglycemic mouse (author's transl)]. The insulin-receptor interaction is impaired in liver, adipocyte and myocardium plasma membranes of the obese hyperglycemic (ob/ob) mouse. A decreased numer of receptors rather than an altered affinity for the hormone appears to be responsible for this defect. Some studies were conducted with respect to the specificity and the reversibility of the degect. 1) It involves the insulin receptor predominantly if not exclusively. 2) It is largely reversible. a) Fasting ob/ob mice for 40 hr or longer periods improved insulin binding significantly. b) Steptozotocin treatment of ob/ob mice also improved insulin binding in liver membranes. c) In fasting and in streptozotocin-treated mice, glucagon binding exhibited a shift opposite to that of insulin, i.e., it was decreased. These data suggest the possibility that the hormone may "down regulate" its own receptor. Although the mechanism(s) of such regulation, if it is verified, remain(s) to be established, similar observations reported with some other peptide hormones and with catecholamines suggest that this phenomenon may be of general interest in endocrine physiology and patho-physiology.


[Androgen binding proteins in human prostate (author's transl)]. Two high affinity saturable binding components were observed in the cytosol of hyperplastic human prostate gland using glyceroltris graident centrifugations having a sedimentation coefficient of 8-10 S and 4-5 S. Binding experiments with different steroids indicate that they correspond to the androgen-receptor protein on one hand and to sex steroid binding plasma protein (SBP) on the other hand. The calculated cytosol receptor concentration was 50 fentomoles/mg protein or 5,000 wites per cell. The SBP concentration, measured by equilibrium dialysis, indicated 8,000 sites per cell which represent 5% of the SBP concentration in the corresponding plasma. Prostate samples were submitted to constant flow organ culture in the presence of 3H-testosterone and increasing amounts of human serum albumin (HSA) and/or SBP. The uptake and metabolism of testosterone were measured and the results suggest that the SBP-testosterone complex could be associated with the prostate cells.


[Androgen binding proteins in the rat prostate: methodological problems and regulation (author's transl)]. Protamine sulfate precipitation is used for the selective and quantitative assay of equilibrium binding constants. The association constant of dihydrotestosterone is 1 X 10(9)1/mole and the number of binding sites is 11.500/cell. The affinity of testosterone is slightly lower than that of dihydrotestosterone, whereas some synthetic androgens have a higher affinity in accordance with their biological activity. Estradiol, progesterone and antiandrogens can displace dihydrotestosterone from its receptor. The occupied cytosolic and nuclear binding sites can be measured by radioimmunoassay. After castration, the nuclear hormone-receptor complexes disappear with a half-life of 3 hours. The cytosol receptor decreases steadily between the first and the fourth day after castration, then increases spontaneously. Testosterone seems to inhibit the degradation and also to stimulate the synthesis of its own receptor. Radioautography shows that the receptor is present only in the epithelial cells.


Evolution of blood cells.

Evolution of blood cells. The most primitive blood cell may have been a protohemocyte which was first involved in phagocytosis and nutrition. When metazoans (sponges) appeared, their "blood" cells, the archeocytes, were phagocytic. In diploblastic cnidarians (e.g. Hydra) blood cells occur in the primitive mesenchyme or mesoglea and are still phagocytic. Although the flatworms and nemertean worms possess a pseudocoelom, a progressive differentiation of several leukocytic types occurred. In coelomate annelids or segmented worms, differentiated cells appeared that distributed food and oxygen, thus erythrocytes evolved in certain marine or polychaete annelids. The Annelida also show differentiation of leukocytes that resemble granulocytes, lymphocytes and monocytes, and cell-mediated immunity that persists throughout later evolution. The Mollusca, Arthropoda, Echinodermata and Tunicata possess essentially the same features as the annelids. The vertebrate blood cells may have evolved from the tunicates via the echinoderm line, the deuterostome invertebrates, although analogous blood cells are present in the annelids, molluscs, and arthropods, the protostomes. The vertebrate plan of blood cell differentiation occurs in two compartments: bone marrow (for both lymphoid and myeloid cells) and lymphoid organs (e.g. thymus, lymph nodes) for lymphocytes.


Thymic lymphocyte origin in the newt Pleurodeles waltlii studied by embryonic grafts between diploid and tetraploid embryos. Thymic lymphocytes origin in the newt Pleurodeles waltlii was studied by embryonic grafts of the gill area between diploid and tetraploid embryos. By this way, either cell size or chromosome number are used as markers. Heterotopic transplantation of the gill area to the ventral region of a host embryo is followed by differentiation of gills and gill-associated structures including thymus. Analysis of cell size and chromosome number shows that in Pleurodeles, thymocytes do not differentiate in situ but originate from blood-borne stem cells that migrate into the thymus anlage.


Ontogeny of the avian thymus and bursa of Fabricius studied in interspecific chimeras. Differences in the structure of the interphase nucleus between two species of birds, the Japanese quail (Coturnix coturnix japonica) and the chick (Gallus gallus) have been used to distinguish cells from different origins in interspecies combinations. This biological cell marking technique was applied to thymus and bursa of Fabricius histogenesis. Using various combinations between components of quail and chick thymic and bursic rudiments, the respective contribution of endodermal epithelium, mesenchyme and blood-borne extrinsic elements to the histogenesis of thymus and bursa was analyzed. It was demonstrated that the whole lymphoid population in these organs is derived from immigrant blood-borne stem cells. Thymus and bursa become attractive for stem cells at a precise stage of their development. In thymus the attractive period last about 24 hours in the quail and 36 hours in the chick. In the bursa of Fabricius the invasion lasts several days in both quail and chick embryos. The mechanisms which control the onset and the interruption of the stem cells inflow have been investigated.


Glucocorticoids and evolution of haemopoietic tissue in the liver of the rat foetus. The number of haematopoietic cells per unit of volume decreases after day 18 and falls rapidly until day 21. On day 21 it is reduced by three quarters as compared to the day 17 value. Maternal laparotomy causes, in the 24 h period following the operation, an anticipated and accelerated decrease. This effect is prevented by previous adrenalectomy. After maternal adrenalectomy, rat foetuses submitted to decapitation in utero, and therefore completely deprived of corticosteroids, do not present in their liver the involution of the haematopoietic tissue normally observed in their intact littermates. Cortisol administration restores the normal involution. These results show the importance of the role played by corticosteroids in the regulation of foetal liver hematopoietic function. The evolutions of proportions and frequencies of the different cellular types composing the haematopoietic tissue studied during normal pregnancy and after either overload or deprivation of corticosteroids, suggest that corticosteroids exert their action essentially at two levels of the maturation process. They decrease the ability of self-maintenance of the precursor cell pool and accelerate the maturation of the polychromatophilic cells, resulting in a progressive exhaustion of the haematopoietic tissue.


Lymphocyte differentiation in the rabbit thymus. Density separation of rabbit thymus lymphoid cells on a continuous and linear Ficoll-Metrizoate gradient resulted in the appearance of three subpopulations of lymphoid cells with peak fractions at densities of approx. 1.067, 1.077 and 1.084 g/ml. The subpopulation of the lowest density appeared to consist of cortisone-resistant cells of various size classes, all characterized by a relative low nucleo-cytoplasmic ratio. Glycerol-induced redistribution of particles within the plasma membrane (IMP) of the lymphoid cells, as revealed with the freeze-fracture technique, was present in the subpopulation of lowest density as well as in thymus lymphoid cells of cortisone-treated rabbits in a high percentage of the cells, and present in a very small percentage of the cells of highest density. It is concluded that an increase in the mobility of plasma membrane components may be a significant feature of T-cell education in the rabbit thymus.


[Results of dermatoglyphic studies in the Roussy-Lévy syndrome]. Dermatoglyphics performed on the three persons with fully developed RLS as well as on those with partial symptomatology revealed a striking agreement as to their pappilary systems and the characteristic deviation from the norm: irregularities on the pattern of the ridges of the finger and toe prints and on the soles and palms, a much reduced "total ridge count" (TRC), predominant bow patterns on the toe prints, grid-and reticularpatterned white lines on the soles and palms, loop proliferation on the soles and special shapes of the transverse palmar crease.


Effect of cortisol in vitro on haemoglobin synthesis by mouse foetal liver. In cultures of foetal mouse liver, collected at various gestational days, the effect of cortisol was investigated on 59Fe uptake into haemoglobin heme and also on the differentiation of the erythroid stem cells or ERC (erythropoietin-responsive cells) into erythroblasts under the action of erythropoietin. Addition of cortisol to the culture medium did not modify in any way the responsiveness of the cell system to erythropoietin but exerted a sustaining action on the rate of haemoglobin synthesis during the second half of the incubation period. This action was restricted to cultures of 15- to 16-day livers and was particularly obvious in erythropoietin-supplemented samples. This selective effect of cortisol likely concerns only the later types of erythroblasts since after 24 h of incubation the polychromatophilic or orthochromatic stages predominate respectively in samples with, or without, erythropoietin. The timing of these results is discussed in the light of previous findings observed in vivo by other authors in the foetal rat.


Effects of antibiotics on metabolism of peptidoglycan, protein, and lipids in Bifidobacterium bifidum subsp. pennsylvanicus. The formation of cell envelope components of Bifidobacterium bifidum subsp. pennsylvanicus was studied by measuring the incorporation of [(3)H]glycine, (14)C-labeled fatty acids, and N-benzoyl-[(14)C]glucosamine into the membrane protein, membrane lipids, and cell wall peptidoglycan, respectively. Inhibition of peptidoglycan synthesis by antibiotics (penicillin G, vancomycin, d-cycloserine, and bacitracin) and by the omission of glucosamine-containing growth factors caused a marked decrease in glycine incorporation into cellular as well as membrane protein, which was accompanied by a considerable enhancement of fatty acid incorporation. The uncoupling of protein and lipid synthesis led to the release of marked amounts of lipids from the cell under these conditions. Arrestment of protein synthesis by antibiotics (chloramphenicol, tetracycline, and actinomycin D) decreased peptidoglycan and lipid synthesis only partially, but did not lead to lipid release. Mg(2+) deficiency of the medium caused about 60% inhibition of growth and lipid synthesis, but protein synthesis and especially peptidoglycan synthesis were much less inhibited. Staphylococcin 1580 arrested the growth and also the synthesis of protein and peptidoglycan. However, the synthesis and turnover of lipids were considerably increased and a release of large amounts of lipids was observed. Peptidoglycan and cellular protein did not show any turnover either during normal growth or after the inhibition of cell wall and protein synthesis.


Cefamandole: in vitro and clinical pharmacokinetics. Cefamandole has a broader spectrum and greater potency than the other cephalosporins. It includes Haemophilus influenzae, most strains of Enterobacter, and many strains of indole-positive Proteus and Bacteroides, with a lower minimal inhibitory concentration for Escherichia coli, Klebsiella, etc. Concentrations of drug in the serum after the parenteral injection of cefamandole exceed manyfold the minimal inhibitory concentrations of over 82% of the bacteria studied. Approximately 65 to 85% is excreted in a biologically active form in the urine. This antibiotic offers advantages of antibacterial effectiveness and at the same time retains the safety of penicillin G and cephalothin in animals.


Biological activity of netilmicin, a broad-spectrum semisynthetic aminoglycoside antibiotic. Netilmicin (Sch 20569) is a new broad-spectrum semisynthetic aminoglycoside derived from sisomicin. Netilmicin was compared to gentamicin, tobramycin, and amikacin in a variety of in vitro test systems as well as in mouse protection tests. Netilmicin was found to be similar in activity to gentamicin against aminoglycoside-susceptible strains in both in vitro and in vivo tests. Netilmicin was also active against many aminoglycoside-resistant strains of gram-negative bacteria, particularly those known to possess adenylating enzymes (ANT 2') or those with a similar resistance pattern. Netilmicin was found to be markedly less toxic than gentamicin in chronic studies in cats, although gentamicin appeared less toxic in acute toxicity tests in mice. The concentrations of netilmicin and gentamicin in serum were compared in dogs after intramuscular dosing, and the pharmacokinetics including peak concentrations in serum were found to be similar.


Comparative nephrotoxicities of netilmicin and gentamicin in rats. The relative nephrotoxicities of netilmicin (Sch 20569) and gentamicin were compared in rats at doses of 30, 60, 90, and 120 mg/kg per day for 15 days. Both drugs caused proteinuria and a decrease in urine osmolality; however, netilmicin produced significantly less changes at all doses than gentamicin. Whereas gentamicin resulted in a decline in creatinine clearance at all doses, netilmicin failed to cause a decline in creatinine clearance. Renal-cortical concentrations of antibiotic at sacrifice were similar in animals receiving either drug. Light-microscopic changes were less severe with netilmicin than gentamicin. Cytosegresomes with myeloid bodies were identified electron microscopically in the kidneys of animals receiving either netilmicin or gentamicin at all doses. Electron-microscopic manifestations were similar. The data indicate that in the rat, netilmicin is distinctly less nephrotoxic than gentamicin.


Antibiotic penetrance of ascitic fluid in dogs. Antibiotic concentrations in ascitic fluid after parenteral therapy may be important in the treatment of peritonitis. We have created ascites in dogs by partial ligation of the inferior vena cava. Ascitic fluid volume was measured at the time each antibiotic was administered. Nine antibiotics were studied in the same three dogs. Antibiotic concentration in ascitic fluid was found to vary inversely with ascites volume. Percentage of penetration (ratio of ascites peak to serum peak x100) ranged from 5.8 to 65% among the drugs studied. Only metronidazole showed a statistically significant higher percentage of penetration than other antimicrobials. Concentrations in ascitic fluid after single doses of cephalothin (15 mg/kg) and the aminoglycosides (2 mg/kg, gentamicin and tobramycin; 7.5 mg/kg, amikacin and kanamycin) did not exceed the minimum inhibitory concentration of many gram-negative rods and may justify the use of higher than usual initial parenteral doses, or possibly initial intraperitoneal administration in seriously ill patients.


Preparation of isolated intestinal villi useful for studying hydrolysis rates of penicillin and cephalosporin esters. A method for the preparation of freeze-dried intestinal villi from hamsters and mice is described. The villi, consisting largely of epithelial cells rich in esterase activity, are useful for hydrolysis studies of penicillin and cephalosporin esters.


Stability of gentamicin in serum.

Stability of gentamicin in serum. Patients' sera were divided into three portions when the initial gentamicin level was determined and were stored at -20, 4, and 25 degrees C in plastic or glass tubes. Gentamicin levels were repeated after 1 and 2 days of storage at the respective temperatures. There was no significant difference in gentamicin levels among portions, except those from a patient in renal failure with high serum concentrations of carbenicillin.


Effect of 9-beta-D-arabinofuranosyladenine 5'-monophosphate and 9-beta-D-arabinofuranosylhypoxanthine 5'-monophosphate on experimental herpes simplex keratitis. Treatment of established experimental keratitis caused by herpes simplex virus with 9-beta-d-arabinofuranosyladenine 5'-monophosphate (Ara-AMP) or 9-beta-d-arabinofuranosylhypoxanthine 5'-monophosphate (Ara-HxMP) showed that the Ara-AMP, in a concentration of 2 or 20%, had a significant effect on the keratitis but that 0.4% Ara-HxMP showed only minimal activity. Ara-AMP was also effective in the treatment of idoxuridine-resistant keratitis. No local toxicity with a high concentration (20%) of Ara-AMP was seen, but the duration of therapy was brief.


Antibiotic synergy and antagonism against clinical isolates of Klebsiella species. Minimal inhibitory concentrations of kanamycin, gentamicin, amikacin, cephalothin, and chloramphenicol were determined in Trypticase soy broth for 70 clinical isolates of Klebsiella species. Gentamicin and amikacin were the most active on a weight basis. Chloramphenicol was more active than kanamycin, and cephalothin was the least active of all. Studies using a microtiter modification of the checkerboard technique were performed to evaluate the comparative activity of the three aminoglycosides in combination with either chloramphenicol or cephalothin. The cephalothin-aminoglycoside combinations demonstrated synergy in >80% of the isolates tested. No antagonism was noted. The chloramphenicol-aminoglycoside combinations showed antagonism in 35 to 45% of the isolates tested. The data suggest that the chloramphenicol-aminoglycoside combinations be used with caution when treating serious infections where Klebsiella is a potential pathogen.


Cephalosporin and aminoglycoside concentrations in peritoneal capsular fluid in rabbits. To study the penetration of antibiotics into peritoneal tissue fluid, a subcutaneous tissue capsule model was modified by implanting multiple, perforated spherical capsules in the peritoneal cavity of rabbits. Capsules became vascularized, encased in connective tissue, and filled with fluid having a mean protein concentration of 3.6 g/100 ml. Capsular fluid was obtained by percutaneous needle aspiration and assayed for antibiotic by the disk plate bioassay technique. Cephalosporins were administered intramuscularly at a dose of 30 mg/kg. Mean peak concentrations of cephaloridine and cefazolin were significantly higher than cephalothin and cephapirin in capsular fluids, but the percent penetration (ratio of capsular mean peak to serum mean peak) ranged from 8.7 to 16.9% and was not significantly different among the cephalosporins. At 24 h the capsular concentration of cefazolin was significantly greater than for the other cephalosporins (P < 0.001). Lower rabbit serum protein binding observed at high in vivo concentrations may have enabled cefazolin to penetrate capsular fluid, but in vitro protein binding studies did not confirm a decrease in serum protein binding at high concentrations within the clinical range. Kanamycin and amikacin showed comparable capsular fluid peak concentrations as did gentamicin and tobramycin. The percent penetration ranged from 15.2 to 34.5% for the aminoglycosides. The only statistical difference was that amikacin penetration was significantly higher than that for tobramycin. Mean capsular concentrations of amikacin, cefazolin, and cephaloridine compared most favorably with the minimum inhibitory concentration of gram-negative bacilli at the dosages used in this study.


Cultivation of Staphylococcus aureus in a synthetic medium of low ultraviolet absorptivity. A new synthetic medium having negligible ultraviolet absorbance is described for growing staphylococci in experiments requiring ultraviolet spectrophotometry of cell-free portions of the culture.


Procedure for expediting determinations of antibiotic susceptibility of gram-negative, urinary tract pathogens. Standardized direct disk diffusion antibiotic susceptibility testing on monomicrobial urine specimens is compared with the Food and Drug Administration method. The direct procedure yields acceptable data and may conserve 24 h in reporting results.


Chemically defined medium for susceptibility testing of antimicrobial agents. A defined medium was developed that supports growth of many of the bacterial and fungal pathogens frequently isolated in clinics.


Medium for use in antibiotic susceptibility testing of anaerobic bacteria. A medium is described which was designed for use in testing the minimal inhibitory concentration of antibiotics for anaerobic bacteria by agar dilution. It contains: Trypticase (1%), Gelysate (1%), yeast extract (0.5%), glucose (0.1%), pyruvate (0.1%), arginine (0.1%), NaCl (0.5%), hemin (5 mug/ml), vitamin K(1) (0.5 mug/ml), agar (1.5%). The medium does not require the addition of blood to support growth of most clinical isolates of anaerobic bacteria.


Preparation of labeled staphylococcal enterotoxin A with high specific activity. Staphylococcal enterotoxin A (SEA) was labeled by the chloramine-T method with 125I to a specific activity of 68 to 300 muCi per mug of SEA and with 131I to specific activity of 8 to 218 muCi per mug of SEA. SEA was partially damaged and aggregated during the labeling and storage. The damage seemed not to be greatly dependent on the specific activity of labeled entertoxin. Crossed immunoelectrophoresis showed two antigenically active and three inactive components in the ascending part of the labeled enterotoxin peak during fractionation by gel chromatography. During storage at 4 degrees C, the antigenic activity of label decreased faster when labeling had been with 131I than when with 125I. The antigenic activity of labeled SEA was lowered remarkably in the ascending part of the protein peak. Greatest release of radioiodine during storage was in the same part of protein peak. According to these results, the most suitable label for radioimmunoassay is obtained from the descending part of protein peak.


Effect of zinc on adenine nucleotide pools in relation to aflatoxin biosynthesis in Aspergillus parasiticus. The adenylic acid systems of Aspergillus parasiticus were studied in zinc-replete and zinc-deficient media. The adenosine 5'-triphosphate levels of the fungus were high during exponential phase and low during stationary phase in zinc-replete cultures. On the other hand, the levels of adenosine 5'-diphosphate and adenosine 5'-monophosphate were low during exponential phase of growth and high during stationary phase. The adenosine 5'-triphosphate levels during exponential phase may indicate higher primary metabolic activity of the fungus. On the other hand, high adenosine 5'-monophosphate levels during stationary phase may inhibit lipid formation and may enhance aflatoxin levels. The inorganic phosphorus content was low in a zinc-replete medium throughout the growth period, thereby favoring aflatoxin biosynthesis. The energy charge during the exponential phase was high but low during the stationary phase. In general the energy charge values were lower because of high adenosine 5'-monophosphate content.


Microbial oxidation and assimilation of propylene. Hydrocarbon-utilizing microorganisms in our culture collection oxidized propylene but could not utilize it as the sole source of carbon and energy. When propane-grown cells of Mycobacterium convulutum were placed on propylene, acrylate, the terminally oxidized, three-carbon unsaturated acid, accumulated. A mixed culture and an axenic culture (strain PL-1) that utilized propylene as the sole source of carbon and energy were isolated from soil. Respiration rates, enzyme assays, fatty acid profiles, and 14CO2 incorporation experiments suggest that both the mixed culture and strain PL-1 oxidize propylene via attack at the double bond, resulting in a C2+C1 cleavage of the molecule.


Sublethal heat stress of Vibrio parahaemolyticus. When Vibrio parahaemolyticsu ATCC 17802 was heated at 41 degrees C for 30 min in 100 mM phosphate-3% NaCl buffer (pH 7.0), the plate counts obtained when using Trypticase soy agar containing 0.25% added NaCl (0.25 TSAS) were nearly 99.9% higher than plate counts using Trypticase soy agar containing 5.5% added NaCl (5.5 TSAS). A similar result was obtained when cells of V. parahaemolyticus were grown in a glucose salts medium (GSM) and heated at 45 degrees C. The injured cells recovered salt tolerance within 3 h when placed in either 2.5 TSBS or GSM at 30 degrees C. The addition of chloramphenicol, actinomycin D, or nalidixic acid to 2.5 TSBS during recovery of cells grown in 2.5 TSBS indicated that recovery was dependent upon protein, ribonucleic acid (RNA, and deoxyribonucleic acid (DNA) synthesis. Penicillin did not inhibit the recovery process. Heat-injured, GSM-grown cells required RNA synthesis but not DNA synthesis during recovery in GSM. Chemical analyses showed that total cellular RNA decreased and total cellular DNA remained constant during heat injury. The addition of [6-3H]uracil, L-[U-14C]leucine, and [methyl-3H]thymidine to the recovery media confirmed the results of the antibiotic experiments.


Nitrate removal in closed-system aquaculture by columnar denitrification. The columnar denitrification method of nitrate-nitrogen removal from high-density, closed  system, salmonid aquaculture was investigated and found to be feasible. However, adequate chemical monitoring was found to be necessary for the optimization and quality control of this method. When methanol-carbon was not balanced with inlet nitrate-nitrogen, the column effluent became unsatisfactory for closed-system fish culture due to the presence of excess amounts of nitrite, ammonia, sulfide, and dissolved organic carbon. Sulfide production was also influenced by column maturity and residence time. Methane-carbon was found to be unsatisfactory as an exogenous carbon source. Endogenous carbon could not support high removal efficiencies. Freshwater columns adpated readily to an artificial seawater with a salinity of 18% without observable inhibition. Scanning electron microscopy revealed that the bacterial flora was mainly rod forms with the Peritricha (protozoa) dominating as the primary consumers. Denitrifying bacteria isolated from freshwater columns were tentatively identified as species of Pseudomonas and Alcaligenes. A pilot plant column was found to behave in a manner similar to the laboratory columns except that nitrite production was never observed.


[Rudimentary stages of the extremities of Scelotes gronovii (Daudin) embryos, a South African Scincidea reptile]. The development of the limbs has been studied in 15 embryos of Scelotes gronovii, found in 8 ovoviviparous females collected at Saldanha Bay, in South Africa, Cape Province. This study leads to the following constatations: In all the young embryos of this species, their appears anlagen of anterior and of posterior limb-buds. The primordia of forelimb-buds retrogress early and disappear, whereas the primordia of hind limb-buds transform into rudimentary limbs which persist in adult. Histological study of the anlagen of fore limb buds establish that 7 somites (S6 to S12, S1 being the first post-otic somite) send ventral processes in the mesoblast of the anlage. These processes follow a sinuous pathway in the limb-bud, and are bent towards the basal cell layer of the somatopleural mesoderm. On the apical part of the limb-bud lie a wholly rudimentary epiblastic ridge, which disappears early. On the apical part of the hind limb-bud an ectodermic ridge is present, well differentiated which transforms soon in an apical fold; and the anlage of the hind limb produced a short conical appendage with short femur, tibia and fibula and one terminal finger. A comparison was made of the main steps of the development of the limbs in three species of Scelotes with rudimentary limbs. Scelotes inornatus, Scelotes brevipes and Scelotes gronovii. In these three species the ectodermal apical ridge of the fore limb-buds is rudimentary or incompletely differentiated; it never transforms into an ectodermal fold and its retrogresses rapidly; and in these species an early arrest of development and an involution of the primordia of the limb-buds occurs. These fact corroborate the anterior observations made on embryos of Anguis fragilis and of Ophisaurus apodus and they strengthen the interpretation postulating that the spontaneous retrogression of the apical ridge is an essential factor in the morphogenetic events involved in the arrest of development of the limb bud in the snake-like Reptiles.


[Ultrastructural study of the piriform cells of the ovarian follicle in 5 reptiles]. In the 5 Lacertilians the cytoplasm of the pyriform cells contains numerous mitochondria, a well developed Golgi apparatus, free ribosomes, glycogen-like particles, annulate lamellae, bundles of microfilaments and microtubules. Both the paracrystalline arrangement of ribosomes in Lacerta muralis and Lacerta viridis and the paracrystalline intramitochondrial inclusions in Anguis fragilis observed during the spring are probably related to hibernation. Each pyriform cell is connected to the oocyte by a cytoplasmic bridge which occurs secondarily between the two types of cells. Therefore the pyriform cells and oocyte constitute a syncytium. It is possible that the cytological similarities between the pyriform cells and the oocyte and the degeneration of the pyriform cells result from this syncytial state. The nucleoi do not have the same ultrastructure in the 5 Lacertilians; differences are noted in the dense fibrillar constituant and the composition of the nucleolus varies. The "paranucleolar body" is present in 4 Lacertilians but absent in Lacerta muralis.


[Stages of development and growth of the thyroid gland and adenohypophysis of the dogfish embryo (Scillium canicula Cuv.), a chondrichthyan fish]. The thyroid morphology, histology, biometry and its anatomical connections are studied during the whole embryonic life in the small spotted dogfish (Scyllium canicula Cuv.). The first differentiation of the gland appears as a thickening of the pharyngeal floor, containing PAS inclusions, at stage 10 mm. The first follicles are apparent at stage 30 mm. The thyroid volumetric relative growth is studied during the whole development and is compared with the absolute embryonic length growth. Volumetric variations of the hypophysis upper parts and ventral lobe are also studied during the whole embryonic life.


[Ultrastructural studies of the evolution of the notochord and dorsal embryonic constituents derived from the mesoderm in the chicken and Japanese quail during teratogenic exposures to parathion]. After experimental expositions to parathion--an organophosphorous insecticide which, in birds, gives rise to elective axial abnormalities--the evolution of the notochord and the different dorsal embryonic tissues issued from the mesoderm was analysed with the chicken and the japanese quail embryos. The abnormalities being settled, the notochord, which has normally developed, exhibited no ultrastructural change. Sclerotomic cells had normally migrated and, at the time when the provertebrae were formed, it was not possible to detect any histological and cytological alterations at their level. At later stages (8-9 days of incubation), the cartilaginous cells of the vertebral body did not show particular sign of an organophosphorous cytotoxicity but their secretory function was weakened, fact that may explain growth retardation. On these observations, it is evident that the tardive cartilaginous injuries cannot determine the axial abnormalities occurring earlier. The main results of this study deal with the cytological alterations of the myotomic elements which appeared to be the earliest and the more important ones. At the beginning (5th day), the myopathic changes were relatively located but they extended to the all cervical area. At 8-9 days, the cervical muscular cells are especially characterised by a lack of myofibrills. In showing only muscular lesions at the first step of teratogenesis, the present investigation set aside a chordal or sclerotomic cells intervention in the teratism induced by parathion. On the other hand, fromthe involvement of the contractile tissue in the formation and in the maintenance of the articulations, it appears that the muscular lesions can offer an embryologic interpretation to the parathion axial abnormalities. The possible effect of parathion in the myopathic evolution by the way of its anticholinesterasic properties is proposed.


[Chronology of development of the genital tract of the calf fetus]. The chronology and the modalities of the differentiation of the genital apparatus were studied in 187 calf fetuses whose insemination age was exactly known (between 32 and 110 days). Male. In the male, the first seminiferous cords form around days 41-42, the interstitial cells appear 2 or 3 days later. From days 60-70 on, the rete testis is made of tubules with an open lumen, which connect the seminiferous cords. The masculinisation of the external genitalia begins as early as day 47 by a rapid increase of the anogenital distance: on day 60, the penis opens under the umbilicus and the scrotum is well differentiated. The regression of the Müllerian ducts starts at the level of their anterior (tubal) part on day 50, when their diameter decreases. From day 58 on, the oviducts become discontinuous and they are almost completely absent by day 63; the uterine horns and the vagina have dissappeared by day 80. The masculinization of the internal genitalia occurs during two phases: 1) between days 56 and 58: the early buds of the seminal vesicles and of the prostate appear, as well as the first differences at the level of the urogenital connections and of the Cowper's glands; a supra-urethral diverticulum develops at the level of the posterior prostatic urethral flexure; the non sexual urethra remains short. 2) After day 70 take place: the differentiation of the epididymides the branching of the seminal vesicles and the stabilization of the Wolffian ducts (after a transitory diminution of their diameter between days 60 and 80). Female. In the female, sexual organogenesis proceeds later than in males. The Müllerian ducts (which show a transitory reduction of their diameter at their tubal level between days 50 and 60) develop steadily after day 60 at the level of their uterine horns and of the vagina; simultaneously the non sexual urethra lengthens rapidly. A suburethral diverticulum, which is absent in the male, develops at the level of the urogenital connections, between days 60-70. The regression of the Wolffian ducts takes place after day 70 when the mesonephros disappear; they first become discontinuous in their median part (at the uterine level) between days 77-80, but important remains of these ducts still persist on day 110, in the posterior part of the vagina. The first primordial ovarian follicles form only after day 100 approximately.


Cutaneous manifestations of disseminated cryptococcosis. Five patients with disseminated cryptococcosis had lesions on the extremities resembling cellulitis, which evolved into areas of blistering and ulceration in three patients. All had underlying disease and were medically immunosuppressed. Disseminated cryptococcosis appears to present with cellulitis or herpes-like vesiculation more commonly than is currently appreciated. India ink preparations of aspirates from areas of cellulitis or Tzanck preparations from blisters may show characteristic organisms, and make possible an immediate diagnosis of cutaneous cryptococcosis. If cutaneous infection is confirmed by performing biopsies and growing cultures, dissemination must be presumed and the patient treated with a full course of systemic antifungal therapy. With increasing awareness of cutaneous involvement, some cases of disseminated cryptococcosis will be diagnosed sooner, leading to earlier therapy and improved prognosis.


Leukocyte migration inhibition assay (LIF) in nickel contact dermatitis. A direct assay using the mixed leukocyte migration inhibition technique for leukocyte inhibition factor (LIF) production has shown that nickel-sensitive subjects can be distinguished from nonallergic controls. The difference between the two groups' migrating indexes was obtained with nickel sulfate in 199 medium at a concentration of 150 mug/ml and without prior coupling to a carrier protein. A concise review of the principal in vitro assays for cell-mediated immunity is included.


Delusions of parasitosis. An approach to the problem. One of the most frustrating problems in clinical dermatology is the patient with a delusion of parasitosis. Over the years, very little realistic and practical advice has been offered to the perplexed dermatologists. We describe an approach that is based on the thesis that dermatologists can and often should treat such patients. By considering each patient as an individual, and by working to build rapport, the dermatologist is in the best position to offer help. We report two illustrative cases and discuss guidelines to treatment.


Cutaneous protothecosis. Successful treatment with amphotericin B. A patient had cutaneous protothecosis because of the alga-like organism, Prototheca wickerhamii. In vitro sensitivity tests showed that the organism was sensitive to amphotericin B, and was treated successfully with this polyene antibiotic. As with treatment of some fungal infections, a clinical response was achieved when therapy with low doses of amphotericin B was given during a short period of time. The basis of the amphotericin B response may have been due to a combination of its immunostimulatory and antibiotic properties.


Herpes zoster. Case report of possible accidental inoculation. A 30-year-old healthy male physician developed grouped, papulovesicular lesions along the dermatomes of T1 and T2 of the left side of his body. The onset occurred two days after he accidentally pricked his right index finger with a needle that had been used to aspirate the acute papulovesicular lesion of a patient with severe herpes zoster. The clinical appearance and dermatomal distribution, the subsequent clinical course, the skin biopsy findings, and the substantial increase in complement-fixing antibody titer to the varicella-zoster (V-Z) virus in the convalescent serum samples are strong evidence for herpes zoster. Although it is generally believed that person-to-person transmission of zoster is rare and that herpes zoster results from the reactivation of a latent varicella virus, the present case suggests that zoster can be acquired from exogenous infection with a V-Z virus, at least in certain circumstances.


Thiabendazole-induced toxic epidermal necrolysis. Toxic epidermal necrolysis (TEN) is a syndrome that may develop from a number of different stimuli, including drug administration, staphylococcal infection, vaccine administration, and blood dyscrasias. A 29-year-old woman developed TEN from the systemic administration of thiabendazole. Although various cutandous adverse reactions have been attributed to the administration of this medication, this is the first case to indicate that orally administered thiabendazole may also produce TEN. Although acutely ill at the time of hospitalization, the patient showed a prompt response to treatment with prednisone.


Thromboatheromatous complications of umbilical arterial catheterization in the newborn period. Clinicopathological study. Severe catheter-related thromboatheromatous lesions were found at necropsy in 33 of 56 infants who had umbilical arterial catheters passed during life. In infants dying within 8 days of insertion of the catheter, varying degrees of thrombosis of the aorta and its major branches were seen. With increasing thrombosis and aging of the thrombus, fatty deposits were seen first within the thrombus, and then in the intima and media. In addition there was evidence of proliferation of medial smooth muscle cells and of disruption of the medial architecture below the thrombus, characterized by the presence of abundant mucopolysaccharide. In infants who survived longer, varying degrees of organization of the thrombus could be traced, leading eventually to raised fibrous plaques with lipid and occasionally calcification. The lesions in the older infants were similar in many respects to experimental thromboatheromatous lesions produced in rabbits, and to some lesions of artheroma occurring spontaneously in humans. A wide variety of embolic phenomena were found, with features suggesting asynchrony of embolic episodes. The presence of thrombotic lesions could not be related to birthweight, Apgar scores at 1 and 5 minutes, age at catheterization, duration of catheterization, underlying disease process, age at death or the presence of hypothermia, acidosis, or anomalies in coagulation tests. There is a need for less hazardous methods of monitoring arterial oxygen tension.


Hyaline membrane disease, alkali, and intraventricular haemorrhage. The relation between intraventricular haemorrhage (IVH) and hyaline membrane disease (HMD) was studied in singletons that came to necropsy at Hammersmith Hospital over the years 1966-73. The incidence of IVH in singleton live births was 3-22/1000 and of HMD 4-44/1000. Although the high figures were partily due to the large number of low birthweight infants born at this hospital, the incidence of IVH in babies weighing 1001-1500 g was three times as great as that reported in the 1658 British Perinatal Mortality Survey. Most IVH deaths were in babies with HMD, but the higher frequency of IVH was not associated with any prolongation of survival time of babies who died with HMD as compared with the 1958 survey. IVH was seen frequently at gestations of up to 36 weeks in babies with HMD but was rare above 30 weeks' gestation in babies without HMD. This indicated that factors associated with HMD must cause most cases of IVH seen at gestations above 30 weeks. Comparison of clinical details in infants with HMD who died with or without IVH (at gestations of 30-37 weeks) showed no significant differences between the groups other than a high incidence of fits and greater use of alkali therapy in the babies with IVH. During the 12 hours when most alkali therapy was given, babies dying with IVD received a mean total alkali dosage of 10-21 mmol/kg and those dying without IVH 6-34 mmol/kg (P less than 0-001). There was no difference in severity of hypoxia or of metabolic acidosis between the 2 groups. Babies who died with HMD and germinal layer haemorrhage (GLH) without IVH had received significantly more alkali than those who died with HMD alone, whereas survivors of severe respiratory distress syndrome had received lower alkali doses than other groups. It is suggested that the greatly increased death rate from IVH in babies with HMD indicates some alteration of management of HMD (since 1958) as a causative factor. Liberal use of hypertonic alkali solutions is the common factor which distinguishes babies dying with GLH and IVH from other groups of babies with HMD. Although the causal nature of this association remains unproved, it seems justifiable to lrge caution in alkali usage.


Biopterin derivatives in normal and phenylketonuric patients after oral loads of L-phenylalanine, L-tyrosine, and L-tryptophan. Plasma biopterin derivatives studied in 10 normal and 21 phenylketonuric children showed a significantly high concentration in the latter group. Biopterin derivatives correlated with plasma phenylalanine concentration, but in normal adults given an oral phenylalanine load the rate of increase with phenylalanine differed from that in phenylketonuric patients. A patient with hyperphenylalaninaemia, not due to phenylketonuria, had an abnormal biopterin derivatives response to phenylalanine distinct from that of patients with classical phenylketonuria. This may be a useful investigation to differentiate some variants of phenylketonuria.


Faecal composition after surgery for Hirschsprung's disease. Diarrhoea and perianal excoriation occur frequently after the endorectal pull-through operation for Hirschsprung's disease. A new method of faecal analysis was performed on 3-day stool collections in 17 postoperative Hirschsprung patients and in 14 normal children, in order to define the faecal abnormality and to establish the cause of perianal excoriation in these patients. Loose stools in postoperative patients were deficient in dry solid content and contained an excess of extractable faecal water. This also had a raised electrolyte concentration, particularly with respect to sodium. Total daily output of faecal water was normal. Formed stools from postoperative patients were also deficient in drysolids but had a normal extractable water content. Excess extractable faecal water, the main abnormality of loose stools in these patients, is the result of abnormal water absorption from the distal colon. Perianal excoriation in these patients is most closely associated with the concentration of sodium in faecal water.


Urinary outputs of oxalate, calcium, and magnesium in children with intestinal disorders. Potential cause of renal calculi. 24-hour urinary outputs of oxalate, calcium, and magnesium have been determined in a total of 62 children aged 3 months to 17 years who fell into the following groups: (i) 16 normal controls, (ii) 3 with primary hyperoxaluria, (iii) 9 with small and/or large intestinal resections, (iv) 9 with untreated coeliac disease, (v) 5 with pancreatic dysfunction, and (vi) a miscellaneous group of 20 children with a variety of intestinal disorders. Taken as a whole, 58% of patients with intestinal disorders had hyperoxaluria, and of these 7% had urinary outputs of oxalate which fell within the range seen in primary hyperoxaluria. The proportion of children with hyperoxaluria in the different diagnostic groups was as follows: intestinal resections (78%), coeliac disease (67%), pancreatic dysfunction (80%), and miscellaneous (45%). 35% of the patients with hyperoxaluria had hypercalciuria, whereas magnesium excretion was normal in all subjects studied. In 2 patients treatment of the underlying condition was accompanied by a return of oxalate excretion to normal. These results indicate that hyperoxaluria and hypercalciuria are common in children with a variety of intestinal disorders, and that such children may be at risk of developing renal calculi without early diagnosis and treatment.


Juvenile nephronophthisis associated with retinal pigmentary dystrophy, cerebellar ataxia, and skeletal abnormalities. A boy aged 9 3/4 years with interstitial nephritis, retinal pigmentary dystrophy, cerebellar ataxia, and skeletal abnormalities is described. The association may be due to a new genetic disorder, since 2 similar cases have been reported.


Abnormal renal functions in cyanotic congential heart disease. Children with cyanotic congenital heart disease had a decreased glomerular filtration rate (71-8 +/- 18-9 ml/min per 1-73 m2) measured by endogenous creatinine clearances, compared with children who had had complete corrective surgery, children with noncyanotic heart disease, and normal children. There was a significant correlation between low glomerular filtration rate and haematocrit values above 50%. Daily urinary sodium excretion was reduced in the cyanotic patients.


Pulmonary eosinophilic gramuloma in a child. The occurrence of pulmonary eosinophilic granuloma in a 3-year-old child is described. She presented with a pneumothorax and typical radiological changes and the diagnosis was confirmed by lung biopsy. There was no objective evidence of improvement after radiotherapy when lung function was assessed by gamma scans. She died suddenly while abroad.


Hormonal changes in thalassaemia major.

Hormonal changes in thalassaemia major. Patients with severe thalassaemia major suffer endocrine and other abnormalities before their eventual death from iron overload due to repeated blood transfusions. The endocrine status of 31 thalassaemic patients aged 2-5 to 23 years was investigated. Exact data were available on the rate and duration of blood transfusion in all of them and in many the liver iron concentration was also known. Although the patients were euthyroid, the mean serum thyroxine level was significantly lower, and the mean thyrotrophic hormone level significantly higher, compared with the values found in normal children. Forty oral glucose tolerance tests with simultaneous insulin levels were performed in 19 children, of whom 5 developed symptomatic diabetes and one had impaired tolerance. Previous tests on all 6 patients were available and some showed raised insulin levels possibly due to insulin resistance. 2 patients had clinical hypoparathyroidism and are described. The parathyroid hormone levels determined by radioimmunoassay in 25 patients were below the mean for the age group in all and outside the reference range in 16. Nonfasting plasma calcium levels were not reduced. Puberty was delayed in some patients. Concentrations of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) measured in urine from 7 girls and 5 boys showed considerable variation. In the boys there was an overall tendency for FSH and LH excretion to be low with regard to age, but with respect to puberty rating FSH exretions were normal or low and LH normal or raised. The girls showed a tendency for LH but not FSH excretion to be raised in relation to puberty rating. The severity of the endocrine changes was related to the degree of iron loading and is discussed in relation to previous work in which the iron loading has rarely been accurately indicated nor parathyroid status assessed.


Functional intestinal obstruction due to deficiency of argyrophil neurones in the myenteric plexus. Familial syndrome presenting with short small bowel, malrotation, and pyloric hypertrophy. In 3 infants functional intestinal obstruction, associated with a short small intestine, malrotation, and pyloric hypertrophy, was shown to be due to failure of development of the argyrophil myenteric plexus, with the absence of ongoing peristalsis. 4 infants with similar clinical features have been described previously, and there is evidence for an autosomal recessive mode of inheritance of this syndrome.


Health needs of preschool children.

Health needs of preschool children. An epidemiological study of disease in a geographically identified population of 250 children is reported. 22% had not seen their general practitioner (GP) at all in the past year, while 20% had seen him four times or more. The vast majority of these visits were because of an infective illness; and developmental and behavioural problems were rarely presented to GPs. 53% of children had not been to hospital since birth, but 11% had been at least four times. Respiratory infections and middle ear disease were the commonest illness reported, and nearly 3% had an infected or discharging ear at the time of examination. 15% of 3 year olds had speech and language problems. 18% of children over 2 years were thought by the examiners to have a behavioural problem, half being assessed as mild, the remainder as moderate or severe.


Family factors affecting child development. In a large, geographically defined population of children a number of family factors in addition to social class, determined by the father's occupation, were recorded by health visitors and school nurses with routine responsibility for these children. The quality of the children in normal schools was assessed in terms of nonverbal IQ and height at the ages of 5 and 10 years, and of behavior as reported by the teacher at the age of 10 years. By analysis of variance the sum of the independent effects of the other family factors greatly outweighed that of occupational social class, except in the case of the IQ at 10 years. The most important of the other family factors was the quality of the mother's care of her child during the first 3 years of life.


Thoracic gas volume in early childhood.

Thoracic gas volume in early childhood. A total body plethysmograph is descirbed which was used to study thoracic gas volume (TGV) in infants and young children from birth to 2 1/2 years, and was suitable for use even in very sick babies. Normal TGV values were obtained in 42 studies of 35 healthy infants and young children, and 16 children with abnormal lung volume are described. TGV correlated well with length, weight, chest circumference, and age in the healthy infants. A low TGV was found in children with respiratory difficulties after cardiac and thoracic surgery, in respiratory distress syndrome of the newborn, and in association with pulmonary infection and chest cage abnormalities. Abnormally high TGV was most frequently seen in infants with small airways disease.


Enterotoxin-producing bacteria and parasites in stools of Ethiopian children with diarrhoeal disease. Enterotoxinogenic bacteria were isolated from 131 (37%) of 354 Ethiopian infants and children with acute gastrointestinal symptoms. Only one of these isolates belonged to the classical enteropathogenic serotypes of Esch. coli. Two colonies from each patient were isolated and tested for production of enterotoxin by the rabbit ileal loop test, the rabbit skin test, and an adrenal cell assay. However, only 38% of the isolated enterotoxinogenic strains were Esch. coli; the others belonged to Klebsiella, Enterobacter, Proteus, Citrobacter, Serratia, and Aeromonas. In 18 patients both isolates were toxinogenic and belonged to different species. The incidence of intestinal parasites was 35% with no apparent correlation to the occurrence of toxinogenic bacteria in the stools.


Estimation of glomerular filtration rate from plasma creatinine concentration in children. The relation between the true plasma creatinine concentration (Pc) and the glomerular filtration rate corrected for body surface area (GFR/SA) was investigated in 108 individuals, and the following formula was derived: GFR/SA (ml/min per 1-73m2SA) = 0-43 Ht (cm)/Pc (mg/100 ml). This formula was tested in a second group of 83 children, and its accuracy and precision was compared to the 24-hour creatinine clearance. It was found to be superior to the creatinine clearance overall, and was as good, even if all results involving suspect 24-hour-urine collections were eliminated from analysis. The formula in SI usage is: GFR/SA (ml/min per 1-73 m2SA) = 38 Ht (cm)1Pc (mumol/l).


Neuroblastoma metastatic to the liver in infants. Four infants are described who presented with rapid enlargement of the liver. This was found to be due to neuroblastoma which had metastasized to the liver; the condition was associated with high levels of urinary vanillylmandelic acid (VMA). In 3 infants the primary tumour was in the adrenal gland and in one it was not identified. One infant died after laparotomy and 2 infants survive disease free with normal VMA levels, one after adrenalectomy and hepatic irradiation and one after a short course of chemotherapy. The fourth patient responded initially to hepatic irradiation and chemotherapy, but relapsed 2 years later with recurrent disease; at the same time the VMA level which had been normal, again rose. The importance of screening for an abnormal VMA level in any infant with a rapidly enlarging liver in order to obtain an early diagnosis is stressed. Careful follow-up, with serial VMA estimations, is essential to detect recurrent disease. The prognosis for some infants with this distribution of neuroblastoma which has metastasized to the liver, but not to the bones or oribt, is good.


Spontaneous perforation of common bile duct in infants. Two infants with spontaneous perforation of the common bile duct are described. One presented with mild jaundice, dark urine, acholic stools, and hydroceles, the other with bilateral inguinal hernia. In both the diagnosis was unsuspected until bile-stained ascites was discovered. Both eventually developed bile-staining of the scrotum. Neither was acutely ill. The 131I-Rose Bengal faecal excretion test showed reduced faecal excretion at 8% and 12% of the injected dose with 16-5 and 17%/dl of the dose being recovered in the ascitic fluid 48 hours after intravenous injection. The ascitic:plasma ratio of isotope at that time was 32:1 and 28:1. Operative cholangiography in both showed a perforation at the junction of the cystic duct and common bile duct with no contrast entering the duodenum. Cholecystenterostomy using a Roux-en-Y loop of jejunum produced a rapid sustained recovery and is suggested as the treatment of choice. This condition should be considered in the differential diagnosis of obstructive jaundice in infancy since early surgical correction is necessary.


Altered connective tissue in children with congenital dislocation of the hip. The umbilical cord was employed as a source of collagen in 10 children with congenital dislocation of hip. The amount of collagen and its solubility were measured in slices of the cords and in the umbilical veins and compared with the values in normal subjects. Both the amount of collagen and its solubility were decreased in children with congenital dislocation of the hip.


Value of cystography in urinary tract infections. Fifty-one children with a bacteriologically proven urinary tract infection had both an intravenous urogram (IVU) and a micturating cystogram. The IVU was normal in 35. Only 6 of these children showed reflux in the cystogram, affecting 7 of the 70 ureters at risk. Since reflux on its own does not cause renal damage, which occurs only with super-added infection, detection of reflux is not important providing the urine is kept sterile. We suggest that cystography be deferred providing the IVU is normal until recurrent infections occur while under hospital care, and, with this policy this unpleasant and sometimes hazardous investigation could be avoided in many children with a single urinary tract infection.


Ultrastructural studies in epidermolysis bullosa hereditaria. IV. Recessive dystrophic types with junctional blistering. (Infantile or Herlitz-Pearson type and adult type). Recessive dystrophic epidermolysis bullosa with junctional blisters includes both the classical epidermolysis bullosa hereditaria letalis Herlitz (Herlitz-Pearson type) and a recently separated more benign adult type. Ultrastructural examination was performed of 13 skin specimens from 3 cases of the Herlitz-Pearson-type and one case of the adult type. Principal ultrastructural changes in involved, intact and experimentally frictioned skin regions, common to all patients, are as follows: In nonseparated areas hypoplasia of the hemi-desmosomes and mild decrease of the tonofibrils are found. Hypoplasia of hemi-desmosomes consists of a marked rudimentary structure of the sub-basal dense plaque and the attachment plaque. Focal widening of the lamina lucida suggesting early blistering occurs exclusively in the areas devoid of hemidesmosomes. In separated areas cleavage always occurs in the plane of lamina lucida, viz. the mode of blistering is junctional. Fragments of basal cells are often encountered still remaining attached to the blister floor ("epidermolytic torn-off phenomenon"). These torn-off portions of basal cells are characterized by relatively rich distribution of hemidesmosomes. Basal cells forming the blister roof frequently show small gaps of basal plasma membrane and rarefaction of the basal part of the cytoplasm, which are thought to be secondary changes. Among the observed alterations, structural defects of hemidesmosomes are considered to play the most important role in the pathogenesis of junctional blisters.


[Skin tests with phytohaemagglutinin (PHA). Correlation with other parameters of cell-mediated immunity (author's transl)]. 100 healthy persons were investigated to find out whether there is a correlation between a lymphocyte stimulation (3H-thymidine incorporation in the lymphocyte transformation test) with phytohaemagglutinin, pokeweed mitogen and concanavalin A and the in vivo reactivity to intracutaneous phytohaemagglutinin application (measured as medium diameter of infiltrates). Further we searched for an influence of positive or negative reactivity to the specific antigens trichophytin, candidin and streptokinase-streptodornase. 1. There is a good qualitative correlation in normal persons between in vitro lymphocyte stimulation by phytohaemagglutinin, pokeweed mitogen and concanavalin A and the medium diameter of infiltrates after intracutaneous application of 2 mug phytohaemagglutinin. Using lower doses of phytohaemagglutinin no such correlation could be found. A quantitative correlation between the medium diameter of infiltrates and lymphocyte transformation by phytohaemagglutinin and pokeweed mitogen cannot be demonstrated. For concanavalin A there is a negative correlation significant at the 99% level. 2. In vivo reactivity to phytohaemagglutinin does not differ in persons with negative and positive reactions to the specific antigens. 3. Lymphocyte stimulation by phytohaemagglutinin, pokeweed mitogen and concanavalin A of persons with positive reactions to trichophytin, candidin and streptokinase-streptodornase is significantly greater than in those with negative reactivity. 4. The results indicate in vivo phytohaemagglutinin testing as a good screening method to judge cell-mediated immunocompetence. The general applicability for differentiation of immunodeficient and immunocompetent patients is to be further investigated.


Glutathione content of human skin carcinomas. Glutathione has important functions in the process leading to the complete neoplastic transformation of the primary cells in experimental carcinogenesis. This study was made to determine the tissue glutathione levels in the human skin carcinomas. The statistical average of glutathione values were measured by the DTNB method as 0.915 +/- 0.073 mg/g wet weight in test group and 0.126 +/- 0.008 mg/g wet weight in control group.


A comparative study of nickel hypersensitivity by the lymphocyte transformation test in atopic and non-atopic dermatitis. In spite of the non-specific mitogenic effect of nickel salts, 3H-thymidine uptake ration (TUR) of nickel-stimulated lymphocyte cultures in nickel-sensitive atopic dermatitis patients and in non-atopic nickel-sensitive patients was significantly higher at 10 mug per ml concentration of nickel salts than in nickel non sensitive subjects (P less than 0.001). A significant difference was also demonstrated at concentrations of nickel salts of 40 mug per ml. The TUR value of nickel-stimulated cultures in nickel-sensitive atopic dermatitis patients tended to decrease in comparison to non-atopic nickel-sensitive patients. Furthermore, the TUR of phytohemagglutinin-p (PHA) stimulated cultures was also lower in atopic dermatitis patients than in non-atopic subjects.


[Ultrastructural investigations on the pathogenesis of hyperplasias and tumors of skin connective tissue (author's transl)]. Results of an ultrastructural investigation of 4 tumors resp. hyperplasias of skin (endometriom, lymphadenosis cutis benigna Bäfverstedt, mastozytom, oral papillomatosis) show that the substance of granules from mastcells and eosinophilic granulocytes influence the composition and synthesis of the groundsubstance and its fibrous materials. Synthesis of the groundsubstance by fibrocyte like cells can be affected by intracytoplasmic uptake of the granules or by liberated granules which can change the composition of the groundsubstance. Fibrocyte like cells can take up the granules direct from the groundsubstance and (or) possibly by intercytoplasmic bridges ("Fursome"). Composition and synthesis of groundsubstance and fibrous elements are related by a feedback system. Therefore an irregular composed groundsubstance cause an irregular synthesis by fibrocyte like cells and vice versa. From that a circulus vitiosus arise that, by our opinion, may contribute to the development of tumors, resp. hyperplasias of skin with essential participation of the connective tissue.


[The leucocyte-migration-inhibition-test in the diagnostic of sarcoidosis (author's transl)]. The positive diagnosis of a sarcoidosis is often difficult since reliable laboratory and radiological parameters are not available. It is therefore often necessary to make the diagnosis with invasive methods. The Kveim skin test has many disadvantages, so that there have been many trials to facilitate the diagnosis by in vitro systems. With a modified and easy handable leucocyte migration technique in agarose (Clausen technique), 30 patients with certified sarcoidosis were tested. As controls served healthy volunteers and patients with various  diseases such as Hodkins disease, Crohn's disease, tuberculosis etc. As antigene a Kveim suspension typ I was prepared according to Chase and was used in a concentration of 100 mug/ml. It could be shown, that leucocytes from patients with sarcoidosis demonstrated a significant inhibition of their migration in presence of Kveim antigene. Besides patients with Crohn's disease in no other group there was a migration inhibition. Therefore this in vitro system allows diagnostic differentiation and helps to find the diagnose which compares well to the results obtained with the Kveim skin test. Since the test is easy to perform and the results are available after 24 h the test is of diagnostic value for the clinical routine.


Method of preparation and application of small quantities of semi-solid topicals. Quantifying with accuracy small amounts of semi-solid viscus materials is critical in some situations. A sample dish and loading device for a micropipette is described which permits efficient and accurate loading and dispensing of microliter quantities. This is especially valuable for dealing with small quantities of radiolabelled viscus materials.


Connective tissue metabolism in culture fibroblasts of a patient with Ehlers-Danlos syndrome type I. Study on connective tissue metabolism was conducted with a patient with Ehlers-Danlos syndrome (E-D) of Type I who visited our institute. The conversion of procollagen into tropocollagen in the medium of cultured fibroblasts was assayed by the chase technique using 3H-proline. The conversion was inhibited in the cultured fibroblasts of the patient. The components of glycosaminoglycans in cultured medium and fibroblasts from E-D were within normal ranges, however, the ratio of glycoprotein to glycosaminoglycans of intra- and extra-cellular fractions of E-D fibroblasts was higher than the normal one. These findings suggest that the insufficient maturation of collagen fiber may be considered fundamental disorders of E-D.


Degranulation of dermal mast cells: effects of fixation and of antidromic nervous impulses on two histochemically identified cell-types. Two types of mast cell can be identified histochemically in the dermis of the rat's external ear. One type is recognized by the binding of concanavalin A (con A) to the cytoplasmic granules (con A-positive cells) while in the other type (con A-negative cells), the granules do not bind con A. The granules in both types are stained metachromatically by toluidine blue. Antidromic stimulation of the great auricular nerve for 2 min results in an increased proportion of degranulating mast cells in the auricular dermis and both types of cell are affected to an approximately equal extent. In discussion of this observation, it is argued that both the con A-positive and the con A-negative mast cells are probably involved in the mediation of vasodilatation due to axon reflexes in injured skin. The proportions of degranulating mast cells determined in histological preparations varied with the fixatives (Carnoy and glutaraldehyde-formaldehyde) used, but the increased degranulation due to antidromic nervous stimulation could be detected after either fixation.


Biosynthesis of prostaglandins in human allergic contact dermatitis. The biosynthesis of prostaglandins by human inflamed skin was studied in 5 patients with allergic contact dermatitis induced by patch tests. Inflamed skin from these patients evidenced an increased ability to synthesize prostaglandins. In the presence of exogenous arachidonic acid in the incubation medium, the activity formed was about 60% greater in inflamed skin than in noninflamed control skin. In prostaglandin E1-equivalents the concentration amounted to 9.19 +/- 2.02 ng/mg protein nitrogen and 5.74 +/- 1.97 ng/mg protein nitrogen, respectively (mean values +/- S.E.). When inflamed skin was incubated without excess of exogenous precursor acids in the incubation medium the activity formed was about 130 times lower and the values were similar to those of non-inflamed skin. Thus, the present results support the view that the prostaglandin system is activated in allergic contact dermatitis, thereby providing a basis for future therapeutic attempts to control this disorder.