Intrarenal platelet consumption in the diffuse proliferative nephritis of systemic lupus erythematosus. 1. Platelet survival and an index of the localization of platelets in the kidney were studied in patients with the proliferative nephritis of systemic lupus erythematosus, either focal or diffuse, and in control subjects. Platelet survival was reduced in patients with proliferative lupus nephritis, more in those with diffuse rather than focal renal involvement. 2. The index of renal platelet localization in patients with diffuse proliferative nephritis suggested an intrarenal platelet consumption not found in other groups. 3. A patient with the classical platelet autoantibody disease, idiopathic thrombocytopenic purpura, also showed reduced platelet survival but localization of platelets was in the spleen rather than the kidney. 4. Intrarenal platelet consumption in diffuse proliferative lupus nephritis may be an epiphenomenon of pre-existing scarring or platelet aggregation secondary to immune complex-formation, which contributes to the progressive sclerosing lesions of this form of nephritis.


A training program in bioanalytical toxicology. Students of diversified backgrounds are taught methods used in bioanalytical toxicology such as TLC, GLC, RIA, EMIT, UV, and spectrofluorometry. Major emphasis is placed on the detection of abused drugs in biologic specimens. It was found that the students with more advanced formal education have learned the lecture theory and instrumentation quicker and in greater detail. However, as far as the ability to work rapidly and accurately, many of the trainees who have had less formal education have shown better ability. Perhaps it could be said that the theory is a science, whereas the work is an art. This art through practice can be better developed by some people regardless of education. On completion of this training, they will be able to help fill the great need for toxicologic technicians.


[Transcription complexes with nucleolar and chromosomal origins in oocytes of Pleurodeles waltlii and P. poireti (amphibia, urodela) (author's transl)]. The method of spreading transcription complexes has been applied to amphibian oocytes of Pleurodeles genus. Complexes of nucleolar origin show a regular and homogeneous organization similar to that described in other materials. The observations add to the interpretation as an amplification of nucleolar DNA and a redundancy of ribosomal cistrons in the two species studied. -- On the other hand, complexes of chromosomal origin display a great diversity. Two main characteristics can be drawn: the existence of several transcription units in a chromosomal organization unit and the possibility to point out a special architecture at the RNP fibril level. Applying a shadowing technique used for isolated molecules is an improvement compared with earlier methods based on PTA coloration.


Premenstrual symptoms and depression in a university population. The correlation between a premenstrual syndrome based on emotional symptoms and primary affective disorder was prospectively studied in 105 college freshmen. Students with premenstrual symptoms were twice as likely as controls to have a history of a serious depressive episodes and twice as likely to have a close family member with depression. Seven percent of women with premenstrual emotional symptoms and none of the controls had an affective episode during the ensuing year. The premenstrual syndrome did not interfere with academic performance and was not associated with any lowering of professional aspirations. The inconsistent results of past research on the relationship between depressive disorder and premenstrual symptoms may have been the consequence of differing definitions in different studies. The final answer on this possible association may result from using a clear definition of premenstrual symptoms based on emotional difficulties and a well defined research oriented criteria for the diagnoses of affective disorder.


[Echocardiographic criteria of asymmetrical hypertrophy of the ventricular septum without outflow tract obstruction (author's transl)]. Asymmetrical septal hypertrophy (ASH) without outflow tract obstruction is a genetic variant of idiopathic hypertrophic subaortic stenosis (IHSS). Without difficulty and risk echocardiography can distinguish both diseases. Comparison in 33 patients with ASH but no outflow tract obstruction and in 29 with IHSS revealed following identical findings: (1) excessive hypertrophy of the ventricular septum when compared with the posterior wall of the left ventricle (asymmetrical septal hypertrophy); (2) decreased systolic motion of the septum; (3) slowed early diastolic closure of the anterior mitral leaflet; (4) relatively small end-diastolic and systolic diameter of the left ventricle. But the important distinction was that, while in IHSS there was a systolic forward movement of the anterior or both mitral leaflets, this not recorded in patients with ASH without outflow tract obstruction. The posterobasal portion of the left ventricle is thickened in patients with IHSS, due to the outflow tract obstruction.


Alterations in steroid and gonadotropin release resulting from surgical stress during the morning of proestrus in 5-day cyclic rats. The purpose of this experiment was to determine whether surgical stress on the morning of proestrus would elicit an early release of gonadotropin from the pituitary. Animals exhibiting 5-day estrous cycles underwent bilateral sham-ovariectomy under ether anesthesia at 0800 h of proestrus. These animals had high levels of progesterone and estradiol following the surgery. These steroids were thought to be adrenal in origin, since animals adrenalectomized at 0800 h of proestrus had low progesterone levels and estradiol comparable to unoperated controls. Subsequently, the sham-operated animals showed high FSH but not LH values at 1300 h, prior to the normal critical period for gonadotropin release. By 1400, the LH surge had begun, and progesterone was again being released. Adrenalectomized and unoperated controls showed no increase in any steroid or gonadotropin measured before 1400 h. These findings suggest that stress-induced release of adrenal estradiol and progesterone, rather than some other consequence of the surgical procedure, during the morning of proestrus, can advance the onset of release of FSH, prior to LH. Ovariectomy at 0800 h proestrus led to a rapid and dramatic increase in FSH but not LH secretion by 4 h after surgery. By 6 h after ovariectomy FSH had increased to six times control values and LH had increased to twice control values. Estradiol remained at control values for 6 h following surgery but 20alpha-hydroxypreg-4-en-3-one (20alpha-OHP) dropped quickly to baseline values. It is possible that a reduction in circulating 20 alpha-OHP may be responsible for the increases in FSH prior to LH in this group, but the absence of other negative feedback factors from the ovary or adrenal may also be involved.


Long-term incubation of rat thymus cells: Cytolytic actions of glucocorticoids in vitro. A system has been developed for long-term incubation of rat thymus cells in sus pension in a chemically defined medium consisting of medium 199 and methylcellulsoe. With this system it is possible to study quantitatively the full time-course of glucocorticoid actions, from the early inhibitory effects on glucose uptake to the slow cytolytic effects. Cells incubated under these conditions preserve their sensitivity to the inhibitory effects of cortisol on glucose uptake for al least 12 h. Glucocorticoids at physiological concentrations lead to cytolysis (measured by reduction in viable cell counts determined by a modification of the pronase-cetrimide method) by 12 h incubation time. The cytolytic effect exhibits well-defined dose-response relationships and specificity for glucocorticoids. By 24 h,cortisol at 10(-6)M consistently reduces the total cell count by about 15%, and the viable cell count by about 40%. Cortisol is active down to 10(-7) M. Dexamethasone is roughly 10 times as active as cortisol, and cortisone completely inactive. Cortexolone is slightly active, but also shows antiglucocorticoid activity by its ability to block the action of cortisol. Under anerobic conditions in Medium 199, no inhibitory effect of cortisol is observed on glucose uptake.


Pinealectomy and sexual rhythm in female rats. The role of the pineal gland in inducing and maintaining the persistent estrus of rats exposed to continuous illumination was examined. Under the cyclic illumination, the pinealectomy or sham-operation revealed no effect on the estrous cycle, although weights of the ovaries, adrenals and hypophysis were slightly but definitely greater in the pinealectomized animals. In rats exposed to the continous illumination immediately after pinealectomy or sham-operation, both groups exhibited the persistent estrous states soon after the change of lighting condition. In these rats, neither the degree of persistent estrus nor the organ weights a autopsy showed any significant difference between the groups. Moreover, the pinealectomy could not alter the incidence of estrous in persistent estrous rats which had been established already by the continuous illumination.


Low molecular weight components of polymers used in packaging. The increasing use of polymers in packaging of foods and drugs focuses attention on the possible chronic toxicity relations of migrants from these polymers to the contents. Such migrants can arise from residues and additives in the polymers from manufacturing processes used in fabrication of packages. The origin and chemical nature of potential migrants, the methods of transfer, and principles involved in development of safety criteria for their regulation are discussed.


Modification of amino groups of human-erythrocyte glycoproteins and the new concept on the structural basis of M and M blood-group specificity. 1. Various kinds of modification of amino groups of M and N blood group glycoproteins abolished their capacity to inhibit rabbit and human anti-M and anit-N sera. 2. The reversible modification of amino groups revealed that M and N blood group activity was restored after the removal of amino-group-blocking residues. 3. Modification of amino groups had an entirely different effect on the reactivity of red cell glycoproteins with Vicia graminea agglutinin. The serological activity of N glycoprotein towards Vicia graminea anti-N agglutinin was unchanged, whereas the weak activity of M glycoprotein towards this anti-N agglutinin was increased to the level of the of N glycoprotein. 4. These results indicate that there is a structural difference between M and N glycoproteins, which resides beyond the oligosaccharide chains. It suggests in turn that M and N blood group specificity is determined by amino acid sequence in the peptide chains of red cell glycoproteins.


Thyroglobulin messenger RNA: translation of a 33-S mRNA into a peptide immunologically related to thyroglobulin. Poly(UC)--Sepharose chromatography of the RNA extracted from a thyroid fraction sedimenting between 800 X g and 27000 X g allows the purification of two RNA fractions amounting each to 1% of the applied material. The first one is loosely bound to the column from which it is eluted at 25 degrees C. It is mainly composed of 16-S and 12-S RNA comprising no poly(A) sequences. This could correspond to mitochondrial rRNA. The second one, which is eluted at 50 degrees C, is poly(A)-rich and represents 33-S and 17--18-S RNA species. The 33-S RNA resists heating at 80 degrees C, suggesting that it is composed of one polynucleotide chain. When injected into Xenopus oocytes, the 33-S RNA specifically promotes the synthesis of a peptide with an apparent molecular weight of 185000 and an apparent sedimentation coefficient of 10-S. This peptide is immunologically related to thyroglobulin and could represent its main precursor. Under the conditions tested it does not polymerize spontaneously into 19-S thyroglobulin, suggesting that assembly of the molecule could require specific, post-translational alterations of the precursor and/or the presence of additional lighter subunits.


Chicken antichromatin antibodies: specificity to different chromatin fractions. Specific complement-fixing and precipitin antibodies were induced in chicken to chromatin from WI-38 human diploid fibroblast. Chicken antibodies were chosen because they present two major advantages with respect to rabbit antibodies: (a) the optimal NaCl concentration for chicken precipitin is 1.5 M, which is also an optimal solvent for chromatin proteins that are insoluble at the lower salt concentration required by rabbit precipitin; (b) chicken antichromatin antibodies require an antigen concentration much lower with respect to rabbit antibodies for saturation at the complement-fixation reaction. These antibodies are species specific and they react at the complement-fixation and precipitin assay not only with the whole chromatin but also with dissociated proteins, albeit at a higher concentration of both antigen and antibodies. The specificity of these antibodies to different fractions of chromosomal proteins and of the chromatin after washing with a solution at a different sodium chloride concentration has been investigated.


Selenium binding to beef-kidney rhodanese. The reaction of beef kidney rhodanese with selenosulfate was studied. The selenium-treated enzyme shows an absorption spectrum with a maximum at 375 nm attributable to a sulfoselenide group. This absorption is bleached by addition of cyanide. After cyanide treatment stoichiometric amount of selenocyanate can be found. The intrinsic fluorescence of rhodanese is quenched by addition of stoichiometric selenosulfate. This effect can be reversed by cyanide or sulfite but not by selenite or glutathione. By comparison with model complexes the selenium-rhodanese intermediate was identified as a cysteinyl-selenium derivative.


Stimulation of proteinase K action by denaturing agents: application to the isolation of nucleic acids and the degradation of 'masked' proteins. Hydrolysis of serum albumin by proteinase K was strongly (greater than 7-fold) stimulated by urea and dodecylsulfate in a dose-dependent manner. With an oligopeptide as substrate, however, proteinase K was inactivated by dodecylsulfate. This indicates that the apparent activation of proteinase K by urea and dodecylsulfate is caused primarily by denaturation of the protein substrates. Although dodecylsulfate inhibited ribonuclease activity in the test-tube completely, it could not prevent RNA degradation during isolation of polysomal RNA, to which ribonuclease had been added, because of the reversible nature of the dodecylsulfate inhibition. Complete protection of RNA, however, was achieved by a combination of dodecylsulfate and proteinase K. The combined action of the detergent and proteinase K was also effective in degrading "masked" proteins in a poly(adenosine diphosphoribose) preparation which could not be attacked by the proteinase alone.


Transport of messenger RNA into different classes of membrane-associated polyribosomes in Ehrlich-ascites-tumor cells. The membrane-bound polyribosomes in Ehrlich ascites tumor cells can be separated into a loosely bound and a tightly bound fraction by means of a high salt treatment. Both membrane fractions as well as the free polyribosomes in the supernatant synthesize about the same set of proteins, suggesting a close relationship between these polyribosome fractions in the Ehrlich cell. Relatively high concentrations of cycloheximide do not prevent newly synthesized poly(A)-containing mRNA from entering the tightly bound polyribosome fraction. Nor had treatment of the cells with puromycin in the presence of cycloheximide, which released about 70% of the nascent chains, any significant effect on the entrance of newly synthesized mRNA into tightly bound polyribosomes. These results suggest that in ehrlich ascites tumor cells nascent polypeptide chains are not involved in the binding of polyribosomes to membranes.


Inactivation of rhodanese by pyridoxal 5'-phosphate. Pyridoxal 5'-phosphate and other aromatic aldehydes inactivate rhodanese. The inactivation reaches higher extents if the enzyme is in the sulfur-free form. The identification of the reactive residue as an amino group has been made by spectrophotometric determination of the 5'-phosphorylated pyridoxyl derivative of the enzyme. The inactivation increases with pyridoxal 5'-phosphate concentration and can be partially removed by adding thiosulfate or valine. Prolonged dialysis against phosphate buffer also leads to the enzyme reactivation. The absorption spectra of the pyridoxal phosphate - rhodanese complex show a peak at 410 nm related to the Schiff base and a shoulder in the 330 nm region which is probably due to the reaction between pyridoxal 5'-phosphate and both the amino and thiol groups of the enzyme that appear reasonably close to each other. The relationship betweenloss of activity and pyridoxal 5'-phosphate binding to the enzyme shows that complete inactivation is achieved when four lysyl residues are linked to pyridoxal 5'-phosphate.


Initiation of protein synthesis in Ehrlich ascites tumour cells. Evidence for physiological variation in the association of methionyl-tRNAf with native 40-S ribosomal subunits in vivo. Binding of methionyl-tRNAf to native 40-S ribosomal subunits is thought to be an early stage in the process of polypeptide chain initiation, and [35S]Met-tRNAf - 40-S-subunit complexes can be isolated from Ehrlich ascites tumour cells following a brief incubation with [35S]methionine. To determine whether this step is subject to modulation by physiological conditions, we have estimated the extent of binding of Met-tRNAf to native- 40S ribosomal subunits in Ehrlich cells under nutritional conditions known to affect the rate of protein synthesis in these cells. Deprivation of either an essential amino acid, lysine, or of glucose, results in a substantial reduction in the proportion of native 40-S subunits which have Met-tRNAf associated with them, and refeeding of lysine to cells deprived of this amino acid partially reverses this effect within 10 min. These effects on the concentration of Met-tRNA - 40-S-subunit complexes are paralleled by changes of similar magnitude in the rate of protein synthesis and in polyribosome profiles. Native 40-S subunits can be spearated by equilibrium density gradient analysis on caesium chloride into two species, with buoyant densities approximately 1.40 and 1.49 g X cm-3. In cells deprived of either lysine or glucose, the radioactivity from [35S]methionine is bound exclusively to the particle of buoyant density 1.40 g X cm-3. In well-fed cells, or in starved cells shortly after refeeding, a significant proportion of the label is associated with a region of the CsCl gradient corresponding to a particle of higher density. The results suggest that the binding of Met-tRNAf to native 40-S ribosomal subunits can be greatly affected by physiological conditions which alter the rate of protein synthesis. This is consistent with a regulatory role for this step in the sequence of reactions involved in initiation of translation.


Circular DNA and rolling circles in nucleolar rDNA from mitotic nuclei of Physarum polycephalum. 1. About 15% of nucleolar DNA (1.712 g/cm3) from Physarum polycephalum displaying maximum hybridization to ribosomal RNA, is composed of circular DNA of 3.9 +/- 0.2 mum contour length or multiples thereof. 2. A portion of these circular molecules (25%) contained linear DNA pieces longer than circumference length. In a small fraction of circular DNA linear pieces, shorter than the unit length, were observed. 3. Most nucleolar DNA, [3H]thymidine-labeled or hybridizable to ribosomal RNA was separable from chromosomal DNA during G2 phase, mitosis and S phase of the cell cycle. 4. Ribosomal DNA content was not amplified during the cell cycle, was unchanged during exponential or stationary growth phase and amounted to about 0.11 -- 0.21% of nuclear DNA in diploid and hexaploid strains of Physarum or 100--200 ribosomal genes per diploid genome.


Relationship of rats' spatial preferences to effects of d-amphetamine on timing behavior. Rats were trained to bar-press on a differential reinforcement of low rate 16 sec (DRL 16) schedule for water reinforcement. On alternate days, rats were tested in the presence or absence of a light which signaled the availability of reinforcement. Rats were allowed to bar-press on either of two levers (left and right). All rats show consistent side preferences. Doses (0.5-2.0 mg/kg) of damphetamine differentially affected performance under signaled and nonsignaled conditions. Performance during the nonsignaled condition was much more sensitive to a drug-induced rate increment and timing impairment than performance during the signaled condition. With increasing drug dosage, under both conditions, side preferences reliably increased, decreased or remained unchanged depending upon the particular pattern of paw usage and the relationship between paw and side preferences. For the nonsignaled condition but not for the signaled condition, baseline rates were related to the strength of side preferences; lower rates and better timing performance wwere significantly correlated with greater preferences. Observations of bar-pressing behavior suggested that stereotyped motor patterns associated with side preferences might be related to mechanisms involved in timing behavior and perhaps, in behavior controlled by internal stimuli generally.


L- and conventional forms of micrococci in the circulating blood of thrombocytopenic patients. The multiplication of Gram-positive Cocci originating from L-forms carried by platelets of autoimmune thrombocytopenic patients, may be attributed to the primary platelet damage enhanced following interaction with bacteria.


[Action of a herbicide (paraquat) on hepatocytes in histiotypic culture]. The toxicant action on hepatocytes of "paraquat" introduced at very low concentrations in an in vitro culture of such cells is here established. Mitoses are slowed down and moreover, the lysosomial system is impaired, thus inducing a decrease in its enzymic reactions and the proliferation of very large vesicles.


Unimpaired fertilization with thymidine-3H-labelled spermatozoa in the mouse. Thymidine-3H-labelled spermatozoa were obtained in the sixth week after injection of males with a single or repeated doses of thymidine-3H, in amounts of 3.5 to 50 muCi per gramme body weight. Mouse ova were fertilized in vivo or in vitro by such labelled spermatozoa. It was found by means of autoradiography that heavily labelled spermatozoa compete successfully with non-labelled ones in performing fertilization. The distribution of relative levels of activity in fertilizing spermatozoa was similar to that in the population of inseminated spermatozoa. The early development of ova fertilized by generally labelled spermatozoa (followed up to the first cleavage) was not impaired. It is concluded that spermatozoa labelled with thymidine-3H in their DNA to a level sufficient for autoradiographic detection may be probably used in the studies of the cytology of fertilization without greater precautions regarding possible radiation-induced artefacts.


Activity and subcellular localization of proteolytic enzymes of boar and rabbit spermatozoa. Comparison of the mode and time course of depolymerization of the gelatin by boar and rabbit spermatozoa obtained from the ejaculates and the female reproductive tract revealed that the proteolytic activity of the spermatozoa was not changed during their passage through the female reproductive tract. In all sperm samples the depolymerization started at the level of the equatorial segment, proceeded to the apical part and digested the substrate around the sperm head. The time course of depolymerization was also the same in all samples. Slight differences were caused by degeneration changes affecting the acrosomal region of the sperm head and accelerating the initial course of substrate depolymerization. The results were confirmed by investigating the state of the acrosomes of individual sperm samples using the silver proteinate method. It appeared that samples with greater numbers of spermatozoa with damaged acrosome begin to affect the substrate a few minutes earlier. The disadvantage of the methods used is that the release of the proteolytic enzymes from the sperm head is probably caused by postmortem degeneration changes of the spermatozoa.


[Electrophysiological studies of the forebrain-limbic inhibitory systems in relation to gonadotropin regulation (author's transl)]. By the earlier studies it has been shown that electrical stimulation of some brain areas such as the medial amygdala and the medial septum increased multiple unit activity (MUA) in the medial preoptic area (MPO) and the arcuate nucleus (ARC) and resulted in gonadotropin release and/or ovulation. On the basis of this evidence, the present study was made to elucidate the effects of stimulation of the forebrain-limbic inhibitory systems on MUA in the MPO and the ARC in relation to the control of gonadotropin release using Wistar female rats. 1) In proestrous rats, electrical stimulation of the basolateral complex of amygdala (1-AMYG), the anterior amygdala (AAA), the hippocampus (HPC), the anterior hippocampus (HIA) and the ventral and lateral part of the central gray matter at the caudal midbrain level (CG) depressed the MUA levels in both the MPO and the ARC in almost of all cases. In the same experiments using ovariectomized and no-primed rats, it was observed that electrical stimulation of the above-mentioned areas was also successful to depress the MUA levels in the MPO and the ARC. But the incidence of inhibition was somewhat lower than that of in proestrous rats. 2) Simultaneous stimulation of the HPC and the 1-AMYG decreased in their inhibitory effects on the MUA in the MPO and the ARC which were apparently observed through the separate stimulation. This was true in combined stimulation of the CG and the 1-AMYG or of the CG and the HPC. It seemed that inhibitory effect on gonadotropin release which was induced by the separate stimulation was cancelled by the combined stimulation. 3) The 1-AMYG and the HPC seemed to form an inhibitory feedback circuit, excitation of a site inhibiting the other. 4) In ovariectomized rats, electrical stimulation of the 1-AMYG or the HPC decreased the MUA levels in the MPO and the ARC after the estrogen injection even in the case that facilitatory effects were observed before estrogen administration. It seemed that inhibitory effects of these inhibitory areas on the hypothalamic activity were enhanced under the dominant influence of estrogen. 5) After an injection of atropine, electrical stimulation of the HPC increased the MUA levels in the MPO and the ARC, while the stimulation of the same sites decreased when no treatment. On the other hand, atropine did not alter the effects of 1-AMYG stimulation on the MUA in the MPO and the ARC. It seems that the forebrain-line inhibitory systems have more important roles in the control of gonadotropin release than it has been assumed, through inhibiting the electrical activity in the MPO and the ARC.


[Human growth hormone and prolactin during pregnancy (author's transl)]. In order to know the secretory behaviors of human growth hormone (hGH) and human prolactin (hPRL) during pregnancy, the following studies were undertaken. Twenty three normal pregnant women of every period of gestation, eighteen women of postpartum and five nonpregnant subjects volunteered for this study. After fasting overnight, these volunteers were placed at complete bed rest, and a fasting antecubital venous blood sample was drawn at 8:00 a.m. Then L-arginine, 30 g, was infused intravenously over a 30 minute period, and venous blood samples were drawn at 30, 45, 60, 90, 120, minutes after infusion. Serum hGH level was detected by hGH radioimmunoassay Kit (Dainabot) and serum hPRL concentration was measured by double-antibody radioimmunoassay system (NIH-NIAMDD). In addition, serum hCS level was measured by hCS-Kobe double-antibody radioimmunoassay system in comparison with the secretory behaviors of hGH and hPRL. 1. Serum hGH, hPRL and hCS concentrations during pregnancy. HGH concentration remained almost unchanged through the course of pregnancy, but hPRL and hCS concentrations increased with the programs of pregnancy. 2. Serum hGH and hPRL concentrations in puerperium. HGH level did not change as compared to that of nonpregnant or pregnant women. HPRL concentration maintained high level in 1-3 postpartum weeks. 3. Effect of arginine on the concentrations of serum hGH, hPRL and hCS during pregnancy. The hGH response decreased, but that of hPRL increased along with the progress of pregnancy. During the arginine loading test there was no significant change in hCS concentration. 4. Effect of arginine on the concentrations of serum hGH and hPRL in puerperium. The HGH response was suppressed at the first week of the postpartum. The response of hPRL was lower than that of late pregnancy. To summarize, hGH and hPRL have some similar biological characters, but there was a difference in secretion pattern of the two hormones during pregnancy. Reserve function of hGH secretion was suppressed, but that of hPRL secretion increased along with the progress of pregnancy. And in the third trimester of pregnancy, the difference of the secretory behavior and secretory reserve function between hGH and hPRL was prominent.


[The effect of menopause on serum levels of calcium and inorganic phosphorus (author's transl)]. Serum calcium, inorganic phosphorus and alkaline-phosphatase were determined in 3,191 women as a part of a multiphasic health testing program. A fasting sample of blood was drawn between 9 and 11 a.m. and the separated serum was applied to a Technicon Autoanalyzer SMA 12/60 and measured. In 527 women, who were found to have no abnormalities on the other laboratory tests or by the physical examination, were the results of determination studied in relation to age and menstrual status. The values obtained from 13,258 men were employed as a control. In regularly menstruating women the serum calcium level was decreased with the advance of age. Once the menstrual cycle had got irregular toward the menopause, the serum calcium level was rapidly increased, reached maximum in 2-5 years after the menopause, and was slightly decreased thereafter. The serum inorganic phosphorus level also varied in a similar attitude. On the other hand both the serum calcium and phosphorus levels in men were gradually reduced with the advance of age and no fluctuation was observed. Alkaline-phosphatase in serum was distinctly enhanced in the postmenopause. These data indicate that the decline in estrogen secretion results in hypercalcemia and hyperphosphatemia and that a prophylactic estrogen therapy may be considered at the early stage of the postmenopause for preventing the increased bone resorption.


Isolation and characterization of 5alpha-pregnane-3,20-dione and progesterone in pepipheral blood of pregnant women. measurement throughout pregnancy. 5alpha-Pregnane-3,20-dione and progesterone were isolated from a pregnancy plasma pool and were identified by using a combination of chromatographic techniques and mass spectrometry. Antibodies to progesterone were obtained in rabbits by immunization with progesterone-1alpha-carboxyethyl-thioether-thyroglobulin. The raised antibodies were of high affinity and one of them cross-reacted (137%) with 5alpha-pregnane-3,20-dione. This property was used to develop radioimmunoassays for measuring circulating levels of both progesterone and 5alpha-pregnane-3,20-dione in pregnancy plasma. The levels of both progesterone and 5alpha-pregnane-3,20-dione increase throughout pregnancy, but a highly significant increase is observed only after the 32nd week of gestation.


Prolonged preservation of Oomycetes and predacious Fungi imperfecti under paraffin oil. Oomycetes and predacious Fungi imperfecti were preserved viable for four years by storage at 22 degrees C under paraffin oil. This method of culture preservation was checked on 52 species belonging to 4 orders and 13 genera.


[Countercurrent distribution of polysulfonated proteins (author's transl)]. The reaction of bovine beta-lactoglobulin AB with reagents 4-(isothiocyanato) benzene sulfonic acid, 5-(isothiocyanato) benzol-1,3-bis(sulfonic acid) and 7-(isothiocyanato) naphthalene-1,3,5-tris(sulfonic acid) is described. The blocking of the epsilon-amino groups of lysine is quantitative. The thus modified protein can be analysed in the countercurrent distribution apparatus and can be split very rapidly at the arginine residues with trypsin.


[Semisynthetic sheep des-A1-glycine insulin (author's transl)]. The synthetic Des-1-glycine-A-chain of sheep insulin as the monomeric cyclic bisdisulfide and native bovine B-chain bissulfonate were reduced together with mercaptoethanol. They combined at pH 10.6 to yield Des-A1-glycine-insulin. This was purified by gel and ion exchange chromatography. The low insulin activity (0.4 - 0.6%) as measured by the fat cell test as well as the change in the CD spectrum indicated that the loss of the N-terminal glycine of the A-chain results in fully inactive insulin. This confirms the results obtained earlier by partial synthesis of Des-A1-glycine-insulin.


Fragmentation of purified mammalian DNA molecules by ultrasound below human therapeutic doses. Because of the increasing medical use of ultrasound, especially in obstetrical diagnosis, it has become important to check its genetic harmlessness. Purified calf thymus DNA in solution was exposed to both therapeutic and obstetrical diagnostic doses of ultrasound. Intensities lower than those used in therapy have a drastic effect on purified DNA in solution, although intensities used in obstetrical diagnosis caused no visible effect.


Statistical evaluation of a new method to detect carriers of phenylketonuria. 40 positive heterozygotes and 43 controls were loaded with 200 mg phenylalanine per kilogram body weight. The aromatic acids excreted 2 hrs after the loading were quantified by gaschromatography. The amounts of mandelic acid (MA), 2-hydroxyphenylacetic acid (2HOPAA) and phenylpyruvic acid (PPA) were used for a discriminatory analysis. The MA concentration alone gives a better discrimination than the statistical analysis.


Familial true hermaphrodism in three siblings: clinical, cytogenetic, histological and hormonal studies. Three affected siblings with the hermaphrodism are described. The propositi showed the following characteristics: male phenotype and gender role, hypospadias, bilateral scrotal ovotestes with palpable nodules, and absence of müllerian structures. The X chromatin was positive and the Y chromatin was negative in the 3 affected subjects. Their karyotype in peripheral blood lymphocytes and in gonadal fibroblasts was 46,XX and no Y chromosome fluorescence was observed. Plasma FSH was elevated in the 2 older patients and plasma LH was elevated only in the oldest. Plasma testosterone was low and plasma estradiol high in the 3 siblings; plasma progesterone was elevated in 2, but normal in 1 sibling. Since some of the clinical characteristics of these 3 affected siblings are not the most common features in the majority of sporadic cases of true hermaphrodism, it is suggested that the presence of all of them may be the first clue for the clinical suspicion of the familial type of true hermaphrodism.


Fibrinolytic activity of allergenic preparations: an artefact due to enzyme contamination. Some of the common allergenic preparations induced lysis of fibrin plates in vitro. Esterolytic and proteolytic activity was found associated with these preparations. All activities were abolished by heat treatment. It is concluded that allergens do not activate human plasminogen and that in vitro fibrinolysis is due to enzyme contamination.


In vitro reaction of antibodies to ragweed. II. Quantitative determination of non-IgE antibodies interferring in a radioallergosorbent test. Immunoglobulin E-rich fractions and IgE-depleted fractions were prepared from individual and pooled sera of patients allergic to ragweed. The IgE-depleted fractions contained antibodies to ragweed associated with the immunoglobulins G, A and M. The IgE-rich fraction contained only IgE antibodies to ragweed, although other immunoglobulins were present in this fraction. The amount of antibodies in these fractions was determined by the radioimmunoassay described in a preceding publication. The inhibitory role of the non-IgE antibodies was examined in an allergosorbent test (RAST) by using various concentrations of the IgE-rich fraction. The RAST was performed using ragweed-coupled cellulose powder as well as paper discs. The IgE antibodies in concentration from 0.017 to 0.069 mug/ml were inhibited by 30-60% in the cellulose RAST by non-IgE antibodies in concentrations from 6.25 to 12.5 mug/ml. The inhibition effect was substantially less pronounced in the paper disc RAST. This would indicate that for diagnostic purposes, especially for examination of sera with high levels of IgG antibodies, it is more reliable to use the disc RAST instead of the cellulose powder RAST.


A narcissistic defence against affects and the illusion of self-sufficiency. A narcissistic defence against affects, unlike isolation, is a defence against an object relationship. Object relations are strengthened by the sharing of genuine affects so that the failure to share feelings or the presentation of false feelings creates distance between the self and other objects. The defence is similar to that of denial in that it entails a modification of the ego's own structure. We have suggested that this modification consists of a precocious but fragile establishment of a sense of self. The defence may occupy a sector of the personality or reflect a more massive structural arrest. When there is this structural arrest, we believe that this narcissistic defence forms the basis for the narcissistic character disorder described by Kohut and the false self of Winnicott. This precocious sense of self leading to an illusion of self-sufficiency may also be found in other disorders, including the borderline patient, but the borderline patient, in contrast, suffers from a failure of internalization which leads to object hunger in contrast to the denial of object need of the narcissistic disorder. We suspect that the environmental trauma that may contribute to the narcissistic disorder is less severe as compared to the borderline states and may consist of the mother's failure to accept the child's separateness and autonomy, resulting in a fear of the mother's intrusiveness. The fear of the maternal object's intrusiveness contributes to the relative inability to form a therapeutic alliance in the psychoanalysis of narcissistic character disorders. The analyst's interpretations are experienced as dangerous, not necessarily because of their content but due to the fear of the analyst's intrusive influence. Our understanding of the means of effecting therapeutic change must be modified in patients with narcissistic character disorders for, in contrast to the 'classical' neurotic, analytic progress is not obtained by means of interpreting the transference neurosis in the context of a working or therapeutic alliance. Although we acknowledge that the psychoanalysis of narcissistic disorders can lead to significant therapeutic gains, such analyses may prove to be interminable if the gains do not also result in the establishment of a transference neurosis and therapeutic alliance.


Ketamine immobilization of northern elephant seals. Ketamine was used as an immobilizing agent to obtain biological specimens from northern elephant seals in their natural habitat. Effective immobilization was achieved with dosages of 1.4 to 6.9 mg/kg of body weight.


An appraisal of naloxone hydrochloride as a narcotic antagonist in the capture and release of wild herbivores. Naloxone hydrochloride was used as the narcotic antagonist during capture operations conducted on 84 specimens of 11 game species in the Kruger National Park, South Africa. It was found that 10 mg of naloxone was sufficient to antagonize wide dosage ranges of etorphine hydrochloride or fentanyl, used in combination with a variety of tranquilizers. The absence of undesirable side effects and the fact that naloxone can be administered without fear of overdosage make it a unique and valuable drug in the capture and release of wild animals.


Dual infection of a white-tailed deer by Dermatophilus congolensis and Alternaria alternata. Infection by both Dermatophilus congolensis and Alternaria alternata was found in a 5 1/2-year-old, female white-tailed deer (Odocoileus virginianus). Encrusted lesions characteristic of dermatophilosis were observed on the hocks, flanks, and back. Giemsa-staining of smears of material from beneath the crusts revealed branching filaments, transversely and longitudinally divided into packets of coccoid cells typical of D congolensis. Hyphae morphologically consistent with those of A alternata were found in methenamine-silver- and hematoxylin-and-eosin-stained sections of tissue from the ears, flanks, and back. Nutrient agar cultures inoculated with tissue from an ear and hindlimb of the deer yielded, respectively, A alternata and D congolensis.


Ataxia, depression, and dermatitis associated with the use of dichlorvos-impregnated collars in the laboratory cat. Ataxia and depression developed in 21 of 50 (42%) laboratory cats wearing flea collars impregnated with 2,2-dichlorovinyl dimethyl phosphate (dichlorvos or DDVP) in a warm dry environment. Five (10%) of the cats died. Whole blood cholinesterase (ChE) activity was significantly (P smaller than 0.001) reduced in all cats and cervical dermatitis occurred in 37 (74%) of them.


Inhibition of development in Dictyostelium discoideum by sugars. Sugars such as glucose, maltose, and trehalose, which are metabolized by Dictyostelium discoideum and which enhance vegetative growth, inhibit the development of the slime mold at concentrations which stimulate growth maximally. They block the acquisition of aggregation competence as well as aggregation. The same sugars also inhibit the degradation of preformed glycogen ribonucleic acid, and protein, which is characteristic of development and which occurs when the amoebas are starved by incubation in dilute phosphate buffer.


Formation of indoleacetic acid by intestinal anaerobes. Indoleacetic acid was produced from tryptophan by only three of 23 intestinal anaerobes studied. Evidence is presented to show that the formation of indoleacetic acid proceeds through the intermediate, indolepyruvic acid, via transamination with alpha-ketoglutarate rather than by tryptamine pathway.


Mutants of Dictyostelium discoideum defective in spore germination. After activation, wild-type Dictyostelium discoideum spores germinate rapidly and synchronously in phosphate buffer as well as in complex medium. Mutants defective in spore germination were isolated and characterized. These mutants (called grm) did not germinate normally in buffer but did germinate in complex medium in the presence of bacteria. One mutant (grm B) swelled normally, but amoebae were not formed. Another mutant (grm F) swelled and germinated poorly in buffer. The members of the third group of mutants (A, C, D, and E) did not swell or give rise to amoebae in buffer.


Inhibition of the assembly and secretion of procollagen by incorporation of a threonine analogue, hydroxynorvaline. Hydroxynorvaline (DL-alpha-amino-beta-hydroxyvaleric acid) was shown to competitively inhibit the activation of threonine and valine when tested with tRNA and synthetases prepared from whole chick embryos. However, the hydroxynorvaline was transferred only to threonyl-tRNA and not valyl-tRNA. The hydroxynorvaline had no effect when tested with other amino acids. The Km for threonine was 25 muM and the Ki for hydroxynorvaline was 181 muM. When fibroblasts from embryonic chick tendons were incubated with [3H]threonine and increasing concentrations of hydroxynorvaline, there was a progressive decrease in the incorporation of [3H]threonine so that 1 mM hydroxynorvaline the incorporation into nondialyzable protein was 26% of the control value. A much smaller decrease in the incorporation of other radioactive amino acids was observed. When the cells were incubated hith [14C]proline and 1 mM hydroxynorvaline, the labeled procollagen containing hydroxynorvaline accumulated intracellularly and very little was secreted. Control experiments demonstrated that free hydroxynorvaline did not inhibit the secretion of unsubstituted procollagen. Although the individual pro alpha chains containing hydroxynorvaline were of normal molecular weight (125,000) and hydroxyproline content, only about 50% of this intracellularly retained procollagen was triple helical within the cell as 37 degrees as measured by sensitivity to pepsin digestion. Also only approximately 50% of the pro alpha chains were disulfide-linked to form triple stranded molecules as compared to greater than 85% linkage in unsubstituted procollagen. We postulate that incorporation of hydroxynorvaline alters the conformation of the propeptide extension sufficiently so that: (a) normal assembly of disulfide-linked, triple helical molecules is reduced and (b) assembled triple helical molecules are not properly recognized by the secretory mechanism.


Purification and comparison of two developmentally regulated lectins from Dictyostelium discoideum. Discoidin I and II. When cells of the cellular slime mold Dictyostelium discoideum differentiate from a nonsocial amoeboid form to a cohesive, aggregating form, they synthesize a lectin-like protein called discoidin, which is present on the cell surface. It is now reported that discoidin consists of two distinct lectins, designated discoidin I and discoidin II, which, although similar in some respects, differ in their electrophoretic mobilities, isoelectric points, subunit molecular weights, amino acid compositions, tryptic peptide maps, the erythrocyte species which they agglutinate, and the sensitivity of their agglutination activity to inhibition by monosaccharides. Furthermore, discoidins I and II differ in their developmental regulation as evidenced by the distinct time courses of their appearance during differentiation.


Magnetic resonance studies of the interaction of Co2+ and phosphoenolpyruvate with pyruvate kinase. Co2+, which activates rabbit muscle pyruvate kinase, competes with Mn2+ for the active site of the enzyme with a KD of 46 muM. Co2+ binds to phosphoenolpyruvate with a KD of 4.1 mM. The structures of the binary Co2+/P-enolpyruvate, and quaternary pyruvate kinase/Co2+/K+/P-enolpyruvate complexes were studied using EPR and the effects of Co2+ on the longitudinal (T1) and transverse (T2) relaxation times of the protons of water and P-enolpyruvate and the phosphorus of P-enolpyruvate. The EPR spectra of all complexes at 6 K, disappear above 40 K and reveal principal g values between 2 and 7 indicating high spin Co2+. For free Co2+ and for the binary Co2+/P-enolpyruvate complex, the T1 of water protons was independent of frequency in the range 8, 15, 24.3, 100, and 220 MHz. Assuming coordination numbers (q) of 6 and 5 for free Co2+ and Co2+/P-enolpyruvate, respectively, correlation times (tauc) of 1.3 times 10(-13) and 1.7 times 10(-13) s, were calculated. The distances from Co2+ and phosphorus and to the cis and trans protons in the binary Co2+/P-enolpyruvate complex calculated from their T1 values were 2.7 A, 4.1 A, AND 5.3 A, respectively, indicating an inner sphere phosphoryl complex. Consistent with direct phosphoryl coordination, a large Co2+ to phosphorus hyperfine contact coupling constant (A/h) of 5 times 10(5) Hz was determined by the frequency dependence of the T2 of phosphorus at 25.1, 40.5, and 101.5 MHz. For both enzyme complexes, the dipolar correlation time tauc was 2 times 10(-12) s and the number of rapidly exchanging water ligands (q) was 0.6 as determined from the frequency dependence of the T1 of water protons. In the quaternary enzyme/Co2+/K+/P-enolyruvate complex this tauc value was consistent with the frequency dependence of the T1 of the phosphorus of enzyme-bound P-enolpyruvate at 25.1 and 40.5 MHz. Distances from enzyme-bound C02+ to the phosphorus and protons of P-enolpyruvate, from their T1 values, were 5.0 A and 8 to 10 A, respectively, indicating a predominantly (greater than or equal to 98%) second spere complex and less than 2% inner sphere complex. Consistent with a second sphere complex on the enzyme, an A/h value of less than 10(3) Hz was determined from the frequency dependence of the T2 of phosphorus. In all complexes the exchange reates were found to be faster than the paramagnetic relaxation rates and the hyperfine contact interaction was found to be small compared to the dipolar interaction. The results thus indicate that the interaction of C02+ with P-enolpyruvate is greatly decreased upon binding to the active site of pyruvate kinase.


Clinical considerations in the choice of materials for orthopedic internal prostheses. The six principal clinical considerations which influence the choice of materials for total joint replacements are given. The necessity for screening tests on potentially useful materials is stressed. These should be carried out on specimens which have been exposed to the sterilizing process, which will be used in clinical practice. The materials should also be investigated after immersion in a physiological solution-bovine serum; following implantation in a large animal. Useful tests are impact testing, and wear studies using pin-on-disk and journal and bush specimens. In the case of total joint replacements, equipment in which joint motions and loadings can be simulated can provide valuable information. The components should also be subject to fatigue testing. There is a need to develop agreed laboratory methods for testing of materials and implants and for the long term follow up of patients.


Aster formation in eggs of Xenopus laevis. Induction by isolated basal bodies. We have assayed various materials for their ability to induce aster formation by microinjection into unfertilized eggs of Xenopus laevis. We have found that purified basal bodies from Chlamydomonas reinhardtii and Tetrahymena pyriformis induce the formation of asters and irregular cleavage furrows within 1 h after injection. Other microtubule structures such as flagella, flagellar axonemes, cilia, and brain microtubules are completely ineffective at inducing asters or cleavage furrows in unfertilized eggs. When known amounts of sonicated Tetrahymena and Chlamydomonas preparations are injected into unfertilized eggs, 50% of the injected eggs show a furrowing response at approximately 3 cell equvalents for Chlamydomonas and 0.1 cell equivalent for Tetrahymena. These results are close to those expected if basal bodies were the effective astral-inducing agent in these cells. Other materials effective at inducing asters in unfertilized eggs, such as crude brain nuclei, sperm, and a particulate fraction from brain known to induce parthenogenesis in eggs of Rana pipiens, probably contain centrioles as the effective agent. Our experiments provide the first functional assay to indicate that centrioles play an active role in aster initiation. None of the injected materials effective in unfertilized eggs produced any observable response in fully grown oocytes. Oocytes and eggs were found to have equal tubulin pools as judged by colchicine-binding activity. Therefore, the inability of oocytes to form asters cannot be due to a lack of an organizing center or to a lack of tubulin. Experiments in which D2O was found to stimulate aster-like fibrous areas in eggs but not oocytes suggest that the inability of oocytes to form asters may be due to an inability of tubulin in oocytes to assemble.


Electron microscopy of synthetic myosin filaments. Evidence for cross-bridge. Flexibility and copolymer formation. Electron micrographs of negatively stained synthetic myosin filaments reveal that surface projections, believed to be the heads of the constituent myosin molecules, can exist in two configurations. Some filaments have the projections disposed close to the filament backbone. Other filaments have all of their projections widely spread, tethered to the backbone by slender threads. Filaments formed from the myosins of skeletal muscle, smooth muscle, and platelets each have distinctive features, particularly their lengths. Soluble mixtures of skeletal muscle myosin with either smooth muscle myosin or platelet myosin were dialyzed against 0.1 M KC1 at pH 7 to determine whether the simultaneous presence of two types of myosin would influence the properties of the filaments formed. In every case, a single population of filaments formed from the mixtures. The resulting filaments are thought to be copolymers of the two types of myosin, for several reasons: (a) their length-frequency distribution is unimodal and differs from that predicted for a simple mixture of two types of myosin filaments; (b) their mean length is intermediate between the mean lengths of the filaments formed separately from the two myosins in the mixture; (c) each of the filaments has structural features characteristic of both of the myosins in the mixture; and (d) their size and shape are determined by the proportion of the two myosins in the mixture.


Centrifugal elutriation: separation of spermatogenic cells on the basis of sedimentation velocity. Various types of cells from the testes of mice and hamsters were separated according to differences in sedimentation velocity by centrifugal elutriation, a counterflow centrifugation technique. Approximately 3 times 10(8) cells, prepared from six mouse testes or from one hanster testis, were separated into 11 fractions in less than two hours as compared to the 4--5 hours required for sedimentation at unit gravity ("Staput"). Fractions enriched in elongated spermatids and spermatozoa (100%), stages 1--8 spermatids (69%) and pachytene spermatocytes (58%) were obtained from mouse testis dispersions. Similarly enriched fractions were obtained from hamster cells. A single fraction enriched in stages 1--8 spermatids (mouse) was prepared in less than 30 minutes. As many as 2 times 10(9) cells were separated in a single procedure. Spermatogenic cells exhibited no evidence of structural damage with trypan blud and phase microscopy, and recovery was essentially 100%. Centrifugal elutriation had no effect on sperm motility or on the plating efficiency of CHO cells.


The inhibition of DNA-dependent RNA sythesis in Yoshida ascites cells by bovine follicular fluid. Follicular fluid aspirated from large cow follicles inhibits endogenous, DNA-dependent RNA polymerase activity in Yoshida ascites cells. The inhibitory component of follicular fluid is probably a protein and appears to affect specifically the activity of the nucleoplasmic polymerase II.


Cell density dependence of the aggregation characteristics of the cellular slime mould Dictyostelium discoideum. We have measured fruiting body density and spore formation efficiency in Dictyostelium discoideum as functions of initial cell density. Experiments were performed on agar made up with distilled water and on buffered agar. Minor differences are seen; these are discussed. The functions show 4 regions of density dependence which can be accounted for by changes in aggregation characteristics with density and changes in the efficiency of spore differentiation. The results are discussed in terms of the relaying mechanism for signal propagation controlling cell aggregation. They extend earlier measurements by Bonner & Dodd and by Hohl & Raper, supply data for a quantitative model of the aggregation process, allow estimates of signal range, and show the importance of entrainment between neighbouring centres in defining aggregation territories.


Identification of Actinomyces viscosus from canine infections. Actinomyces viscosus is a gram-positive, non-acid-fact, facultative, catalase-positive, filamentous, or diphtheroidal microorganism. It was isolated from six canine infections during a period of 1.5 years. The organism was cultured from exudate and flaky granules aspirated from infectious granulomas and empyemas. All cultures grew well aerobically and anaerobically with the addition of 10% carbon dioxide. They fermented lactose, produced catalase and acetylmethylcarbinol, reduced nitrates, hydrolyzed aesculin, and did not produce gelatinase or urease. These physiological characteristics distinguish A. viscosus from other morphologically similar organisms.


Reference equine antisera to 33 human adenovirus types: homologous and heterologous titers. Equine antisera to human adenovirus types 1 to 33 were prepared and evaluated by hemagglutination-inhibition and serum neutralization tests. Detailed data on the potency and purity of the immunizing antigens were tabulated as one means of evaluating the antisera. Most of the 52 hemagglutination-inhibition and 25 serum neutralization major or minor heterotypic responses among the equine antisera were observed at similar levels in previous studies with rabbit antisera and appeared to represent genuine antigenic relationships among the human adenoviruses. Equine antisera to human adenoviruses 1 to 33 and a similarly packaged normal horse serum served as lots of fully tested sera for definitive typing of isolates and as reference standards for evaluating other antisera.


Evaluation of turkey erythrocyte hemagglutination assay for the detection of hepatitis B antigen. A recently described hemagglutination test for hepatitis B antigen (HBsAg) using turkey erythrocytes coated with horse antibody to HBsAg purified by affinity column chromatography was evaluated on a comparative basis with 100 HBsAg-positive and -negative serum samples. The turkey erythrocyte hemagglutination test (TEHA) was found to be less sensitive than radioimmunoassay (RIA) but gave far better results than counterimmunoelectrophoresis. Quantitative titration of HBsAg in serial dilutions of the samples appeared to be more reliably performed by TEHA than by RIA. TEHA is a simple and sensitive technique for the detection of HBsAg and may offer several practical advantages over RIA.


Rapid conversion of Histoplasma capsulatum, Blastomyces dermatitidis and sporothrix schenckii in tissue culture. A simple method for positive identification of Histoplasma capsulatum, Blastomyces dermatitidis, and Sporothrix schenckii is given. Primary tissue cultures of guinea pig peritoneal macrophage were inoculated with the mycelial phase of each organism and after 24 h the cells were stained and observed microscopically. The characteristic yeast phase could then be observed allowing for positive identification.


Dorsal column nuclei projections to the cerebellar cortex in cats as revealed by the use of the retrograde transport of horseradish peroxidase. The existence of a cerebellar projection from the dorsal column nuclei (gracile and cuneate nuclei, DCN) has been proposed on electrophysiological grounds but questioned when studied with neuroanatomical techniques. The retrograde transport of horseradish peroxidase (HRP) has been used for the present study and provides anatomical evidence of a DCN-cerebellar pathway. In adult cats, 1 to 6 mul of 30% HRP were injected in pars intermedia of the anterior lobe (lobules IV-V), in paramedial lobule and in vermis of the anterior (lobules IV-V) and of the posterior lobe (lobule VII). After survival of 24 to 48 hours, all animals were perfused with a double aldehyde mixture and serial 40 mu sections through the medulla oblongata were incubated for visualization of HRP. In all cases, medullary nuclei known to project to the injected cortical regions of the cerebellum contained HRP-positive neurons mainly ipsilateral to the injection (e.g., external cuneate nucleus) or mainly contralateral to it (e.g., inferior olivary complex). Following ipsilateral injections in either the paramedian lobule or the pars intermedia, HRP-positive neurons in the cuneate nucleus were concentrated in its rostral portion where multipolar cells with radiating dendrites predominate. In contrast, none of the clusters region, in the caudal part of the cuneate nucleus, displayed HRP-positive granules. In cases in which the anterior vermis was injected a few labelled cells were present in the rostral part of the gracile nucleus but not in the clusters region of this nucleus. No labelling of DCN neurons was evident after posterior vermis injection. To compare the distribution of cells contributing to the DCN-cerebellar pathway with that of thalamic relay cells in the DCN, 0.5 to 3 mul of 30% HRP were injected in the nucleus ventralis posterolateralis of the thalamus in another series  of cats. Contralateral to the thalamic injection, labelled cells were concentrated in the clusters region of the gracile and cuneate but rostrally in these nuclei they were scattered among unlabelled neurons. The preferential location in the DCN of cells which project to the cerebellum and of cells which project to the thalamus stresses the heterogeneous organization of these nuclei along the rostrocaudal axis. Further, the results indicate that regions of the DCN which have been distinguished on the basis of cytoarchitectonics (Kuypers and Tuerk, '64) and of afferents (Rustioni, '73, '74) differ also in their efferent projections.


[Urogenital tuberculosis in 1974].

[Urogenital tuberculosis in 1974]. Urogenital tuberculosis, although often forgotten, remains a common and serious disease. Over a period of 10 years (1963 to 1973) 52 patients were seen and treated on our unit. There were 4 deaths all due to intercurrent disease, and 24 patients, i.e. half the survivors list one kidney owing to the tuberculosis. These crude figures, when one considers that they were mostly middle-aged adults, give an idea of the serious nature of urogenital tuberculosis in 1974.


Maternal influences on ontogeny of suckling and feeding rhythms in the rat. To assess the effects of both maternal and endogenous factors on the ontogeny of nocturnal feeding in the rat, day and night weight gain was determined in the sighted or blind litters of sighted or blind litters of sighted or blind dams and in young weaned at 15 days of age. Sighted dams impose a diurnal milk-intake pattern on their young. The nocturnal pattern of solid-food intake begins at 19 days of age. Blind litters also display nocturnal feeding when kept with sighted dams, but only until 35 days of age.


Decline of maternal behavior in the virgin and lactating rat. Sensitized virgins and postpartum lactating mothers, both exhibiting maternal behavior, were given donor litters that increased in age by 1 day, for 28 days, starting at the onset of maternal behavior. Each day females were tested for maternal behavior with pups 4-8 days old: Maternal care (i.e., nursing/crouching, retrieving, nest building and licking) and maternal withdrawal, rejection, and prevention of nursing were recorded.  After the ninth day, females were also tested with the progressively older pups from 10 to 28 days of age with which they were living. Virgins and lactating mothers showed generally similar patterns of maternal care although some differences were found, and they declined in maternal behavior toward the older pups in a similar manner. Maternal behavior did not decline in tests with younger pups. The results are interpreted as supporting the hypothesis that the decline as well as the maintenance of maternal behavior postpartum is nonhormonally mediated.


Fenitrothion. I. Study of mutagenic activity in rats. The mutagenic activity of fenitrothion was studied in rats given 0,10,40 or 80 ppm of fenitrothion in the diet. The study combined the dominant lethal test with cytogenetic analysis of chromosomal aberrations. Dominant lethal mutations were investigated: 1. by their so-called tentative determination in single mating in P-to F3 generation males and females following 200 days exposure; 2. by assessing the effect of the agent at individual stages of spermatogenesis, with F2 and F4 generation males having been exposed for 100 days and mated to unexposed females for 10 weeks. Chromosome aberrations were analyzed in the bone marrow of F2 generation males following 200-days exposure and F3 generation (males) following 500-day exposure to a dose of 80 ppm. Negative results were obtained in all experiments in relation both to dose and generation. Hence fenitrothion is not considered to be a substance with a mutagenic activity. The metodical advantages of the proposed combination of reproduction and mutagenic-activity studies of an agent for toxicological evaluation are discussed.


Vaccination of cattle against Q-fever.

Vaccination of cattle against Q-fever. Formalin-killed phase I C. burneti organisms containing the protective antigenic component were used for vaccination of heifers against Q fever. None of the vaccinated heifers contracted Q fever, whereas 8 out of 12 (66.6%) control, non-vaccinated heifers became infected when exposed to infection for 3 months in the breed of naturally infected dairy cows. In the vaccinated cattle C. burneti was detected immediately post partum neither in the placenta nor in the colostrum. No shedding of C. burneti in milk has been found so far during 3 years of post-vaccinal observation period. For detecting Q fever antibodies in the blood of vaccinated as well as naturally infected animals MAR was found more sensitive than CFR.


Nucleic acid content and nucleotide composition of DNA in leptospirae. The contents of DNA and RNA were studied in 9 strains of leptospirae and the composition of nitrogen bases of DNA in 20 strains of pathogenic and saprophytic leptospirae. It has been found that leptospirae contain a high per cent ration of nucleic acids. According to the nucleotide composition, the family of Leptospira as a whole belongs to the AT-type. According to overall guanine and cytosine contents, the investigated strains of pathogenic leptospirae fall into 3 groups and differ from the seprophytic strains. With respect to the limited number of the investigated strains, the DNA nucleotide composition of leptospirae can be used as a supplementary biochemical criterion in the classification of leptospirae.


Yersinia enterocolitica as a causative agent of septicaemia. The authors describe the first case of isolation of Yersinia enterocolitica from blood in a septic infection in Czechoslovakia. They also isolated a strain of Y. enterocolitica from an inguinal tumour in the same patient. Both strains belonged to biotype 4, serotype 0:3 and phagotype 8 and were non-pathogenic for guinea pigs and hares.


Litter sizes of Ha(ICR) mice: a preference for even numbers? The frequency distributions of litter sizes at birth for eight sublines derived from the same Ha(ICR) mouse stock with or without selection for large litters have their modes at even numbers of young. Each of the eight sublines produced a majority of even-numbered litters. An upward integral shift relationship among the medians of the frequency distributions was established. These findings are interpreted to mean that, in response to continued plus selection, ovarian and uterine performance of the Ha(ICR) bipartite reproductive system tends to be maximal bilaterally, which results in an excess of even-numbered litter sizes.


Comparison of confirmation methods for hepatitis-B antigen and the nature of false-positives detected by 125I-immunoglobulins. Immunologic specificity associated with solid-phase direct radioimmunoassay (RIA) using guinea pig antibodies for HBsAg detection (Ausria-125TM) was examined further with physicochemical techniques. The RIA reactivities which could not be neutralized by wide-spectrum human anti-HBs serum appeared to be physically distinct from true HBsAg particles; hence, they were truly false-positive, resulting from immunologic cross-reactions. In order to ensure the full advantages of using this highly sensitive RIA test, a confirmatory test using specific human anti-HBs for neutralization is therefore required to distinguish the true- and false-positives. The basic RIA technique is a two-step procedure. Two basic confirmation procedures, namely, a first-step neutralization and a second-step neutralization, were investigated in depth to assess their efficacy and practicality for confirmation. Both procedures were effective for confirmation purposes; however, the first-step neutralization procedure failed to confirm some high-titered HBsAg samples unless these samples were appropriately diluted. The second-step neutralization method did not require dilutions of any test samples.


Mobitz type I atrioventricular block in the ventricular specialized conduction system. A patient with a history of multiple syncopal episodes had electrocardiographic findings of Wenckebach type of second degree atrioventricular block and left bundle branch block. He was thought to have intermittent complete heart block. His bundle recordings demonstrated the unusual occurrence of Mobitz type I block localized to the ventricular specialized conduction system. Based on the findings of this case and those of previous case reports, it is recommended that electrophysiologic studies should be performed on all patients with Mobitz type I atrioventricular block who also have bundle branch block.


[In vitro differentiation of cartilage from cephalic neural crest of Pleurodeles waltlii Michah (author's transl)]. Parts of cephalic neural crest of Pleurodeles waltlii have been cultivated in vitro alone or in explants containing other cellular components. It was hoped that these experiments would establish under what conditions cephalic neural crest cells can differentiate into chondroblasts. It has been demonstrated that this differentiation is possible only under the influence of the dorsal mesoderm and foregut endoderm. It appears that neural crest cells are relatively pluripotent and progressively differentiate in response to local environmental factors.


Marker chromosome analysis of two mouse chimaeras. Chimaeric mice were obtained by injecting embryonic (CBA/H-T6 X PDE)F1 cells into PDE blastocysts. Three of 15 young were overt chimaeras. One female and one male chimaera survived to adulthood and after completion of test breeding, which demonstrated chimaerism in the germ cells of both, they were killed for study when aged 32 and 33 weeks respectively. Chromosome spreads were scored for the presence or absence of the T6 marker chromosome in direct preparations from bone marrow, spleen, thymus, lymph nodes, Peyer's patches, corneas, gut epithelium and testes. Preparations from monolayer cultures of skin, kidney, ovary and gut from mitogen-stimulated blood cultures were scored in the same way. Both components of the chimaeras were identified in every one of 53 specimens studied, some of which, such as single lymph nodes, corneas, and segments of gut, may not have contained more than 10(5) proliferating cells. This result complements published evidence for fine-grained mixture obtained in morula-aggregation chimaeras by other methods and implies extensive cell movement during embryogenesis. Results obtained from the lymphomyeloid tissues show a clear partition into two groups in respect of the proportions of host-type to donor-type cells indentified. The one group consists of bone marrow, thymus and Peyer's patches, the other of spleen and lymph nodes. This result would be most simply explained in terms of two distinct stem cell pools and appears to conflict with the currently favoured hypothesis of a single stem cell pool for the whole lymphomyeloid complex located in bone marrow. Four groups of factors may, however, modify the relative representation of the two components in different lymphomyeloid sites: (1) The magnitude of embryonic founder populations. (2) Limited recruitment from the stem cell pool in post-natal life. (3) Variable size of clones produced by individual stem cells. (4) Differential cellular behaviour determined by genotypic differences.


Proportionality in the pattern of differentiation of the cellular slime mould Dictyostelium discoideum and the time of its determination. A quantitative measure of the proportionality of the pattern of cell differentiation is obtained by separating populations of fruiting bodies into stalks and spores and determining the ratio of their dry weights. The effect of incubation temperature on the proportion of a population which becomes stalk cells is determined. The time of determination of this proportion is then indicated by the time in the developmental sequence at which a temperature shift fails to alter it. The results show that the temperatures of growth, aggregation and migration have no effect on the pattern of differentiation and that temperature alterations during early culmination alter the pattern of differentiation. This result demonstrates that the pattern of differentiation is not determined during the migrating slug stage, and it is suggested that the axial inhomogeneities seen in the slug are not directly related to the terminal pattern of differentiation of the fruiting body as has been previously suggested.


Macrocyst genetics in Polysphondylium pallidum, a cellular slime mould. The discovery of two mating types in the cellular slime mould Polysphondylium pallidum is reported. Two developmental mutants produced in strains of opposite mating type but which do not proceed past the aggregation stage of development are  capable of producing macrocysts. These macrosysts were viable and 5 to 10% germinated after 6 weeks of storage. When the macrocyst progeny were cloned, several classes of non-parental phenotypes were recovered.


The hydrogenation of unsaturated fatty acids by five bacterial isolates from the sheep rumen, including a new species. Five strictly anaerobic bacteria able to hydrogenate unsaturated fatty acids were isolated from sheep rumen. One was characterized as Ruminococcus albus, two as Eubacterium spp. and two as Fusocillus spp., one of which is named as a new species. The Fusocillus organisms were able to hydrogenate oleic acid and linoleic acid to stearic acid, and linolenic acid to cis-octadec-15-enoic acid. The R. albus and the two Eubacteria did not hydrogenate oleic acid but converted linoleic and linolenic acids to a mixture of octadecenoic acids; trans-octadec-II-enoic acid predominated but several isomeric cis and trans octadecenoic acids were produced together with isomers of non-conjugated octadecadienoic acids. The intermediate and final products of hydrogenation by each organism were compatible with the results from mixed rumen bacteria.


Numerical analysis and computerized identification of the yeast genera Candida and Torulopsis. Numerical analysis of the published standard descriptions of 104 species of Candida and 48 species of Torulopsis suggested that the number of species should be reduced to 78 and 33 respectively. Four examples are noted of closely matched species differing in ability to assimilate nitrate; these species may be combined as nitrate-variable species. Although the ability to form pseudomycelium is the only important difference between the genera, only three examples were noted of Candida and Torulopsis species sufficiently closely matched to be regarded as synonymous. Close relationships with species of the perfect genera were more common. A system, derived from the results of the analysis, is described for computerized identification of species of both genera.


Soil fungistasis: role of the microbial nutrient sink and of fungistatic substances in two soils. Sensitivity of conidia of Cochliobolus victoriae to fungistasis decreased markedly following incubation on moist sand for at least 1 h. Germination was greater on Conover loam or on sand being leached with water than on an alkaline clay loam soil known to produce a volatile fungistatic substance. Evolution of 14CO2 began within 3 min after [14C]glucose was applied to the soils; the rate of 14CO2 evolution was faster with Conover loam. Germination of Thielaviopsis basicola conidia per unit of glucose remaining in agar discs initially containing 0-1% glucose, was lower for discs incubated on the clay loam soil than on Conover loam, and was greatest on a bed of sand undergoing aqueous leaching. Germination of ascospores of Neurospora tetrasperma and conidia of C. victoriae was suppressed on discs of washed, Purified Agar or polyacrylamide gel incubated on or over the clay loam soil, but no suppression resulted when discs were incubated on Conover loam. Extensive aeration of either soil did not remove its fungistatic effect. Fungistasis in Conover loam appears to be caused primarily by nutrient deprivation, whereas volatile fungistatic substances may play a major role in the clay loam soil.


Identification of Chromobacterium violaceum: pigmented and non-pigmented strains. The classification and, therefore, identification of Chromobacterium violaceum has been based upon its ability to produce a violet pigment. Although the organism may yield non-pigmented variants when subcultured on artificial media, the isolation of non-pigmented strains from pathological tissues or from nature had not been reported. With a method established for the identification of C. violaceum regardless of violet pigmentation, non-pigmented strains were isolated from nature. The presence of non-pigmented strains of C. violaceum in nature is of significance to taxonomy and clinical bacteriology. Pigmentation cannot be held as an essential characteristic of the definition of the genus Chromobacterium and gives credence to the suspicion of Sneath (1960, 1966) that the genus is not a natural one. Non-pigmented strains may have been isolated from clinical material but wrongly identified as belonging to other genera of non-pigmented Gram-negative bacilli and regarded as not being pathogenic.


Correlation of virulence with secretion in vitro of three wall-degrading enzymes in isolates of Sclerotinia fructigena obtained after mutagen treatment. Correlation and regression analyses were carried out between the virulence (expressed as growth rate in apple fruits) and the secretion in vitro of three host wall-degrading enzymes by 119 isolates of Sclerotinia fructigena, most of which had been obtained following exposure of conidia to N-methyl-N'-nitro-N-nitroso-guanidine. Virulence was found to be significantly correlated (P less than 0-01) with alpha-L-arabinofuranosidase, but not with pectin esterase or, where enzyme interdependence had been statistically eliminated, with polygalacturonase. Approximately 35% of the total variability in virulence could be accounted for in terms of the three enzymes.


The fine structure of Eadie's ovals isolated from sheep rumen. The structure of two strains of the Gram-negative rumen organism, Eadie's Oval, was examined with the electron microscope. Despite their large size, their fine structure indicated that they were bacteria. They had a cell envelope consisting of two membranes separated by a dense layer which could be solubilized by lysozyme. They possessed characteristic bacterial flagella, and lacked internal organization with ribosomes and DNA-like material dispersed throughout the cytoplasm. The outer membrane was corrugated and each strain had a characteristic pattern of corrugations. One strain had sheathed flagella, the other did not. Both strains were coated with fibrils up to 660 nm long, but which apparently contracted to give an unusual cross-banded layer when treated with lysozyme.


Frog virus 3 replication: electron microscope observations on the terminal stages of infection in chronically infected cell cultures. An examination of BHK, CEF, and FHM cells chronically infected with frog virus 3 has been made by scanning and transmission (thin section, freeze fracture, and surface replica) electron microscopy. With minor differences the pattern of virus development is similar in all three cell line. Virus particles were detected in cell nuclei which subsequently became degenerate very late in infection. Three inclusions were associated with frog virus 3 cytoplasmic foci of infection; lamella structures, extensive microtubule formation (in BHK and FHM cells), and linear crystalline structures. The last two structures may play a role in creating or maintaining the cell rounding c.p.e. revealed by scanning electron microscopy. Very late in infection most BHK and FHM, but not CEF, cells are stripped of the plasma membrane. Replicas of frozen fractured BHK cells featured cytoplasmic foci of infection, budding at the plasma membrane, and showed that at early times when virus is detected in the nucleus, the nuclear membranes are intact and morphologically unaltered. Budding at the plasma membrane was better resolved by scanning and as surface replicas. This demonstrated that sparse to profuse localized budding occurred. Frequently virus particles were located singly, or as multiples, at the end of, or along, cytoplasmic protrusions which occur both on the body of the cells and at the cytoplasmic/coverslip 'interface'.


Angiography in urinary bladder tuberculosis. Angiography was performed in 6 cases of bacteriologically and microscopically verified tuberculosis of the urinary bladder. As a rule, general or locally accentuated hypervascularization in the bladder wall could be demonstrated in the arterial phase but no venous abnormalities were observed. The circulation time was usually not changed. Thus the angiographic appearances were non-specific. Angiographically it is impossible to differentiate between tuberculous cystitis, cystitis of other genesis or even tumour of the bladder.


Lymphography as a guide to prognosis in malignant testicular tumours. Lymphography was performed in 130 patients with malignant neoplasm of the testes. The patients were treated by irradiation, supplemented by chemotherapy when lymph node metastases were present. The survival rates of the various pathologic subdivisions were analyzed according to the lymphography. The following 5-year survival rates were observed: seminoma: normal lymphography 94%, pathologic lymphography 69%; teratocarcinoma: normal lymphography 86%, pathologic lymphography 14%; embryonal carcinoma: normal lymphography 64%, pathologic lymphography 33%.


The heat production associated with the passage of a single impulse in pike olfactory nerve fibres. 1. A study has been made of the temperature changes associated with the passage of a single impulse in the non-myelinated fibres of the pike olfactory nerve. 2. The initial heat occurs in two phases: a burst of positive heat, followed by an evolution of negative heat. The positive and negative heats, and the net initial heat, are temperature-dependent. 3. At 0 degrees C the measured positive initial heat is 44.2 mucal/g.impulse; and the corresponding negative initial heat is 48.9 mucal/g.impulse. There is thus a net initial heat that is negative, of about 4.7 mucal/g.impulse. 4. The positive heat has a positive temperature coefficient, being increased by a factor of 1.86 when the temperature is rasied from 0 degrees C to 10 degrees C. 5. The negative initial heat also increases when the temperature is raised, but less than the positive initial heat. As a result, the net initial heat tends to become positive at higher temperatures. 6. Because of temporal dispersion of the action potential over the face of the thermopile, the observed temperature changes are smaller than those that occur at a single point in the nerve close to the stimulating cathode. The value of the positive heat at 0 degrees C corrected for temporal dispersion is estimated to be about 62 mucal/g.impulse: the corresponding value for the negative heat is about 67 mucal/g.impulse. 7. All records were analysed in terms of only two phases of initial heat (one positive, one negative). No analysis required four phases; but it is unclear whether this finding reflects a true absence of four phases, or merely the inability of the recording equipment to resolve them. 8. The positive heat seems to be derived from two sources. First, there is a dissipation of the free energy stored in the membrane capacity. Secondly, there is an evolution of heat corresponding with a decrease in entropy of the membrane dielectric with depolarization.


[Role of the silent period preceding the rapid voluntary movement (author's transl)]. It has been shown that there is the silent period preceding the rapid voluntary movement. The present paper is designed to find the mechanisms of this inhibitory phenomenon and to investigate their role in the voluntary movement. The following results were obtained: 1. It was found that the silent period before the movement was observed not only in reaction trials to visual stimulus but also in voluntary trials without stimulus. 2. The silent period were simultaneously recorded from the rectus femoris, the vastus lateralis and the vastus medialis by upward jumping movement. This finding suggested that this inhibitory phenomenon was a change in excitability of motoneurons innervating the quadriceps. 3. When a subject extended his elbow and knee at the same time, the silent period before the movement could be seen in both triceps brachii and vastus medialis. 4. These results suggest that the upper center sends some inhibitory discharge to the motoneurons before the movement. It is supposed that the silent period preceding the voluntary movement plays a major role in the mechanisms of motor control.


Factors governing the periodic activation of supraoptic and paraventricular neurosecretory cells during suckling in the rat. 1. One hundred and thirty-two antidromically identified paraventricular (PV) and supraoptic (SO) neurones were studied during milk ejection evoked by the suckling of the young, in thirty-four lactating rats anaesthetized with urethane (1.1 g/kg I.P.). The spike activity of these neurones was analysed to determine some of the factors governing their orthodromic activation. Changes in spike activity were compared with the amount of oxytocin released from the neurohypophysis, as measured by the rise in intramammary pressure at milk ejection. 2. Sixty-five units, from both the PV and SO nuclei, displayed a characteristic burst of accelerated activity 12-18 sec before milk ejection. With a suckling stimulus of nine or ten pups, these neurosecretory responses were both regular in their occurrence (recurring evey 4-8 min) and uniform in character, i.e. all responsive cells accelerated to 30-80 spikes/sec for 1-4 sec. Two units deviated from this pattern and both were recorded in animals which failed to display a detectable milk ejection...


A hydrophobic binding site in acetylcholinesterase. The dissociation constants have been determined and compared for a series of reversible, noncovalent inhibitors of eel acetylcholinesterase that are structurally related to the very potent inhibitor, 1,2,3,4-tetrahydro-9-aminoacridine (THA). It is concluded that there exists on the enzyme protein, closely adjacent to the anionic subsite, a conformationally flexible, hydrophobic area which tends readily to assume a near planar form. The dimensions of this area are unknown, but it is adequate in size to fully accomodate THA. It is this area, acting conjointly with the adjacent anionic subsite, which provides the attraction for THA and related inhibitors. Uv absorbance maxima and pKa vlaues are reported for many of the compounds.


Analogs of bradykinin with restricted conformational freedom. Three analogs of bradykinin have been synthesized which bear an alpha-methyl group in the place of an alpha proton at position, 4, 5, or 8. Such analogs possess restricted conformational freedom and are of interest for three reasons. (1) They may provide information about the receptor-bound conformation of the peptide. (2) They may provide a route to antagonists of the native peptide. (3) They may be degraded slowly by proteolytic enzymes. None of the analogs described here antagonized the action of bradykinin, but one exhibited tissue specificity and decreased pulmonary inactivation in the rat.


Studies of platinum complex inhibition of leucine aminopeptidase. The inhibition of swine kidney leucine aminopeptidase by N-alkyl-substituted etyhlenediamine liganded dihaloplatinum chelates has been investigated. The rate of this inhibition at 37 degrees is considerably less than that for the platinum tetra- and hexahalo complexes and also the ethylenediamine dihalo chelates. Only mixed inhibition was observed in these studies. For the time studies used here little if any inhibition occurred at room temperature. L-Methionine and L-Ala-L-Met did not reverse the platinum chelate inhibition nor did they prevent it.


Mutagenic activity of furylfuramide on cultured mouse cells. Effects of furylfuramide (FF) on cultured FM3A cells a C3H mouse mammary carcinoma cell line, were examined. FF inhibited the growth at 10(-4.5) M, and provoked enlargement of cellular, nuclear and nucleolar size, cytoplasmic vacuolation and granular aggregation of chromatin. Chromosome preparation demonstrated severe aberrations in nearly 90% of mitotic plates after 24 and 28 hour treatment. The changes included gaps, breaks, exchanges and fragmentations, but were at chromatid level. Time course study of the incorporation of radioactive precursors showed the gradual but remarkable inhibition of 3H-thymidine uptake whereas 3H-uridine and 3H-leucine uptakes were fairly maintained. Thus, it is suggested that FF induced the block of G1 phase and the delay of S and/or G2 phase. According to the results of alkaline sucrose gradient analysis of cell DNA, breakage of the treated cell DNA was induced and the induced breakage was recovered after the incubation without FF. The capacity to induce 8-azaguanine-resistant mutant cells by FF was shown remarkably high.


[Diagnosis and treatment of macular lesions in multifocal chorioiditis (presumed histoplasmosis) (author's transl)]. The authors describe the clinical picture of "presumed histoplasmosis" with the typical disciform mascular lesion which is found in a increasing number of Dutch patients. Although systemic histoplasmosis is not endemic in the Netherlands, the authors feel they are dealing with the same entity. The therapy is elucidated, especially (Argon laser) photocoagulation of favourably situated subretinal neovascularisations. The importance of adequate coagulation technique and frequent (fluographic) follow-up studies in view of possible complications is stressed.


[Lead poisoning in childhood: a silent epidemic in the slums of the United States (author's transl)]. While leadpoisoning has been with mankind since the dawn of civilization, only in recent years this disease was recognized as a serious public health problem. It has been estimated that more than half a million children are suffering from lead intoxication in the U.S.A. In most instances this disease is caused by eating lead containing peeling paint or plaster crumbs which are abundantly found in run-down neighborhoods of the slums. Children, often unsupervised, develop a craving for these toxic substances and their parents are usually unaware of the inherent dangers of such activities. Education of parents, early recognition and appropriate treatment of affected children and -- most important -- eradication of slum living supported by legislative action and governmental financial aid comprise the ingrediences to a rational approach to overcome the presenting dilemma in childhood lead poisoning.


[Studies on the incidence and clinical importance of the valve independent, arrested hydrocephalus (author's transl)]. Between 1964 and 1974 193 infants up to the age of 2 years were treated for progressive hydrocephalus by means of a ventriculo-atrial shunt. Long term follow-up studies were possible on 148 children. Among these 94 have functioning and 54 non-functioning valves. The criteria for the non-function were based on either plain X-rays of the skull and thorax (disconnection of the catheters, distal end of the cardiac catheter above D 2), direct punctures of the antichamber and contrast medium studies of the diversional system. 35 children with non-functioning valves were normal in their mental and motor development, 9 were retarded according to their primary brain damage, 10 patients showed signs of chronic increased intracranial pressure. These results suggest, that about one fourth of the valve treated patients develop a state of valve independency.


[The unspecific reaction of inflammation in rickets due to lack of vitamin D (author's transl)]. 10 children (3--18 months old) with Vit. D Deficiency Rickets were investigated by Rebuck skin windows for their ability to elicit unspecific inflammation before and after therapy. Differences to a control group (13 children) were statistically significant. In the acute phase of the disease beginning of inflammation is retarded, reduction of polymorphonuclear leucocytes in favour of mononuclear cells is less then in controls, the absolute quantity of exsudation is reduced too. After therapy inflammation is intensified and accelerated, but exsudation is still different from control children qualitatively and quantitatively. These findings add new facts to the data already known on impaired defence mechanisms in rickets.


[Enterobacter-osteomyelitis in two neonates (author's transl)]. Two boys aged up to 2 weeks suffered from enterobacter-sepsis. In both cases osteomyelitis developed in spite of treatment with Gentamycin or Gentamycin combined with Chepazolin. Both children were, taking accont of the risks, then treated with Chloramphenicol (100 mg/kg body weight/24 hours) and the first patient also, for a short time, with tetracyclin. In the second patient we saw a marrow depression dependent on Chloramphenicol and its dosage which disappeared rapidly, when the drug was withheld.


[4 cases of congenital afibrinogenemia (author's transl)]. Report on 4 members of one family with afibrinogenemia. 2 of them died of cerebral hemorrhage at the age of 10. The 2 surviving children are 8 and 10 years old. Plasmafibrinogen was determined in 69 members of the family. Only in the parents of 2 patients was plasmafibrinogen found to be slinghtly reduced. But in other members of the family fibrinogen was low. In our cases, too, the ADP induced aggregation of thrombocytes in plasma obtained with citrate was weaker than normal.


The effect of intact and lysed leukocytes on murine tumor growth and host survival. The inhibitory effect of sensitized lymph-node cells on transplanted tumor growth in the mouse is well documented. The efficacy of peripheral lymphocytes is disputed. Lysed lymphocyte extracts ("transfer factor"), although effective in transferring several types of delayed hypersensitivity in man, have had equivocal results in animals, particularly in tumor immunity. Fifty 129SV mice were sensitized over 1 month with multiple injections of a transplantable tumor that had arisen in the strain as a spontaneous testicular teratoma. These lymphocyte donor mice were sacrificed and lymphocytes pooled. Half of the lymphocytes were given intact to the 25 mice of experimental group A. Remaining lymphocytes were lysed and given to the 25 mice of group B. The control group C of 25 mice received intact lymphocytes from unsensitized donors. All mice were then challenged with 10(6) tumor cells subcutaneously. Two weeks after challenge no animals in the treated groups (A and B) had palpable tumors, whereas 32% in the control group C had palpable tumors. After 3 weeks 25% of group A, 42% of group B, and 63% of group C had tumors. Tumors ultimately grew in 60% of group A, 76% of group B, and 88% of group C. The effect of treatment on rate of tumor growth after appearance was variable, showing inhibition in some and enhancement in others. The ability of intact, sensitized lymphocytes and lymphocyte extracts to confer relative tumor immunity was demonstrated.


Immunological status as a criterion for operability of genitourinary cancers. Delayed hypersensitivity reaction of 116 consecutive cases of urologic cancers were studied by skin tests. Ninety-one percent of those who were operable or free of diseases at least six months had positive skin reaction. All patients with inoperable or recurrent bladder, kidney, or testis tumor had negative skin reaction. All patients with operable prostatic cancer and 52% of those with inoperable prostatic cancer had positive skin reaction. Skin testing could provide useful data regarding the extent of tumor and operability. It is proposed that skin testing be included in routine preoperative evaluation of urologic cancers.


[Formation by Actinomyces malachitospinus sp. nov. of a physiologically active substance that stimulates zygote formation in Mucorales]. A new species Actinomyces malachitospinus sp. nov. is described. It has spiral catenulate spores with spines, a gray aerial mycelium, and a green colonial mycelium, and contains no soluble pigments (including melanoid pigments). The strain of Act. malachitospinus INMI 217 produces a physiologically active substance that stimulates the formation of zygotes in Phycomyces blakesleeanus.


[Morphological and biosynthetic activity of Trichothecium roseum in relation to the polynuclear conidia of the fungus]. The effect of the number of the nuclei on the morphological and synthetic variability was studied with the conidia of Trichothecium roseum. Treatment of suspensions of the conidia with 0.5--1.0 percent solutions of colchicine during two hours increased the number of the nuclei in the conidia, induced the formation of giant colonies, and augmented the ability to produce antibiotics. No reliable differences have been found in the levels of proteolytic activity between the control strains and the strains treated with colchicine.


Effects of low-dose X-irradiation and age of females on intra-uterine death in mice. Pilot-tests were carried out on the effect of low X-ray doses (0, 2, 4, 8, 12, 16 and 32 rad) and age of females on intra-uterine death. The tests included the effects of various ages of females at exposure, of fractionated irradiation and pre-partal exposure. Females and males were taken from our inbred CBA strain. Virgin females, as well as females that had produced some litters, were used. From the results it is obvious that the age of the females at the time of mating had a considerable effect on the death rate (varying from about 8% in young females to 25-30% in old ones). The low-dose X-ray exposure does not seem to have had any detectable influence on the rate of intra-uterine death.