Effect of heparin modification on its activity in enhancing the inhibition of thrombin by antithrombin III.

Studies were conducted to determine the effect of modifying specific functional groups of heparin on its antithrombin III-enhancing activity. The derivatives employed were heparin methyl ester, heparinylglycine and N-desulfated heparin. The carboxyl-modified derivatives increase the rate of inhibition of thrombin by antithrombin III, although not to the same extent as heparin. N-Desulfated heparin is devoid of any activity. Heparin methyl ester is more potent than heparinylglycine in activating antithrombin III, as exhibited by its immediate effect on the thrombin-fibrinogen reaction. However, heparinylglycine is the more effective of the two, in increasing the rate of thrombin deactivation by antithrombin III. The results indicate that although free carboxyl groups of heparin are not crucial for its binding to antithrombin III, they are important for the combination of the latter with thromobin. In contrast, N-sulfates are critical for the interaction of heparin with antithrombin III.


Vitamin A receptors. II. Characteristics of retinol binding in chick retina and pigment epithelium.

Gel filtration studies demonstrate that retinol receptors of chick retinal and pigment epithelial cytosols are (1) of very similar nature (2) of small molecular size (about 18000 daltons) and are different in character from serum proteins. Citral inhibits the binding of [3H]retinol to the retinal 2 S receptor. Retinol acetate competes with retinol for binding to 2 S receptor in both retina and pigment epithelium whereas retinol palmitate is an effective competitor only in the pigment epithelium. Dithiothreitol maximizes 2 S binding in retina and pigment epithelial cytosol; its absence does not lead to receptor aggregation however. A limited number of high affinity binding sites (2 S receptor) appear to be present in retina and pigment epithelium. A 5 S binding species is also present in pigment epithelium; it is similar in character to [3H]retinol binding in serum and may arise from serum contamination of the pigment epithelial preparation. Binding affinity in retina is high with possibly two classes of retinol binding sites present of KD about 1 - 10(-9) and 4 - 10(-8).


Synthesis of a mannosyl phosphoryl polyprenol by the cellular slime mold Dictyostelium discoideum.

A membrane fraction isolated from the cellular slime mold Dictyostelium discoideum was incubated with GDP-[14C]mannose and found to catalyze the incorporation of [14C]mannose into an endogenous acceptor to yield a product with the chemical and chromatographic properties of a polyprenol phosphate sugar derivative. These results suggest that D. discoideum can synthesize a mannosyl phosphoryl polyprenol.


[Photoregulation of L-phenylalanine ammia-lyase: an immunochemical approach].

The photoregulation of L-Phenylalanine ammonia-lyase (PAL) is studied by immunochemical methods. We used a partly purified L-Phenylalanine ammonia-lyase: F1 light fraction, the corresponding inactive one provided from dark-grown cotyledons: F1 dark fraction and the antisera specific of these two fractions. The complete absorption of PAL activity from F1 light fraction with the anti-F1 light immune serum shows the antigenicity of PAL and the specificity of this serum for all forms of PAL present in F1 light fraction. The presence of an inactive L-Phenylalanine ammonia-lyase in the 36 h dark-grown cotyledons suggested by preliminary results of absorption is conformed by showing that less PAL activity is precipitated from the fraction F1 light by a same amount of IgG anti-F1 light when F1 dark fraction is added. This result is explained by a competition between active and inactive forms of PAL for the IgG extracted from an immune serum specific for F1 light fraction. By measuring the absorption of PAL activity from FO fraction (crude extract) obtained from 18 h, 36 h and 48 h light-grown cotyledons when increasing amounts of IgG anti-F1 36 h light are added, we demonstrate the presence of at least two isozymes A and B, the synthesis of B being shifted in time in comparison to A.


Some mechanical property considerations of reconstituted collagen for drug release supports.

Reconstituted collagen, having been shown previously to be biodegradable and to have low immunologic activity, has been further evaluated here as a vehicle for drug delivery. Mechanical stress-strain (Young's Modulus), mechanical abrasion resistance, and steroid (medroxyprogesterone) diffusion rates have been determined for collagen films subjected to nine different chemical preparative treatments. The treatments fall into three categories: formaldehyde crosslinking, chrome tanning, and a combination of formaldehyde and chrome. Based on our conditions, the formaldehyde films have been found to have higher Young's Moduli and lower diffusion release rates of medroxyprogesterone acetate (MPA) than the untreated films. The largest chrome complexes apparently open up the collagen matrix to yield higher diffusion release rates than the untreated film. The Young's Modulus is 2--3 times higher for the chrome films that the untreated film but the abrasion resistance is generally substantially less. Combination chrome/formaldehyde treatments yield film properties between those of the two separate treatments.


Cellobiase from Trichoderma viride: purification, properties, kinetics, and mechanism.

Three distinct cellobiase components were isolated from a commercial Trichoderma viride cellulase preparation by repeated chromatography on DEAE cellulose eluting by a salt gradient. The purified cellobiase preparations were evaluated for physical properties, kinetics, and mechanism. Results from this work include: 1) development of one step enzyme purification procedure using DEAE-cellulose; 2) isolation of three chromatographically distinct, yet kinetically similar, cellobiase fractions of molecular weight of approximately 76,000; 3) determination of kinetics which shows that cellobiase hydrolyzes cellobiose by a noncompetitive mechanism and that the product, glucose, inhibits the enzyme, and 4) development of an equation, based on the mechanism of cellobiase action, which accurately predicts the time course of cellobiose hydrolysis over an eightfold range of substrate concentration and conversions of up to 90%. Based on the data presented in the paper, it is shown that product inhibition of cellobiase significantly retards the rate of cellobiose hydrolysis.


Phenotypic modifications in amino acid profiles of cell residues of Candida utilis and Enterobacter aerogenes.

Glucose-limited chemostat cultures of Candida utilis were cultivated at various pH levels (3.0-7.5), temperatures (15-37.5 degrees C), dilution rates (0.006-0.42 hr-1), and with one of two nitrogen sources (NH+4 or NO-3). Enterobacter aerogenes was also cultivated in the chemostat under nitrogen and phosphorus limitations. The amino acid profile of total cell protein is expressed as the content of each amino acid relative to the sum of all amino acids recovered ater acid hydrolysis. Cell residues obtained after hot trichloracetic acid extraction display small variations in amino acid profile. Some of these variations correlate with the growth rate at satisfactory levels of statistical significance. In C. utilis, the correlations cover increased levels of lysine, arginine, and leucine and decreased levels of serine and glutamic acid with increased "reduced dilution rate" (D/Dc). In E. aerogenes, increased levels of lysine and arginine and a decreased level of glutamic acid correlate with increased dilution rate. The directions of most of these correlations and the extents of those pertaining to lysine and arginine are consistent with the change predicted to occur simultaneously in the relative level of the ribosomal protein group.


[Effect of neurotropic substances on sleep disorders caused by electrical stimulation of the hypothalamus in cats].

The total period of sleep decreased as a result of the REM-sleep deficite following rage reaction induced in cats by the electrical stimulation of the hypothalamus. Haloperidol (1, 2, 3 mg/kg), diazepam (0.5, 1 mg/kg), nitrazepam (1, 6 mg/kg), glutetymide (10, 30, 60 mg/kg), pentobarbital (5, 15, 30 mg/kg) failed to eliminate sleep disturbances induced by rage reaction; lithium hydroxybutyrate (100, 150 mg/kg), dimedrol (1.5, 6 mg/kg) and imipramine increased the total sleep time on account of the slow wave phase; sodium hydroxybutyrate (100 mg/kg) normalized the electrophysiological pattern of sleep, decreasing the REM-sleep latency and the number of waking cats, and increasing the total REM-sleep time and the number of REM-sleep episodes.


[Effect of lithium chloride on ethanol uptake by rats].

A selective uptake of ethanol by presenting its 5% solution as the only source of fluid was elaborated in rats for 2 months. It was found that lithium chloride injected intravenously in a dose of 35 mg/kg twice per 24 hours for 14 days depressed the ethanol preference causing a motivation inversion whose mechanism was associated with the changes in the activity of the hypothalamic centres of the neuroendocrine regulation. A possibility of lithium salts in the therapy of chronic alcoholism is discussed.


[Influence of psychotropic substances on elaboration of a defensive conditioned reflex in experimental neurosis].

Neurosis resulting in alteration of the motor-defense conditioned reflex development occurred in rats subjected to a long-term action of a negative-emotional factor--neighbour-rat's reaction to pain. Benactyzine, diazepam, chlordiazepoxide, and sodium hydroxybutyrate improved the formation of defense conditioned reflex during this form of neurosis. Trioxazin, trifluoperazine, and amphetamine produced no such effect. The proposed model of experimental neurosis can be used to reveal the tranquilizing activity of psychotropic drugs.


[Primary and secondary immune response in animals following spontaneous loss of tolerance].

Mice subjected to tolerogenic treatment by sheep red blood cells (SRBC) and cyclophosphamide were immunized at various intervals (from 1 to 8 weeks after treatment) either by a single injection of 5 X 10(8) SRBC or by a double-injection of 1 X 10(6) SRBC. In control mice both immunization methods proved to be equally successful. In the experimental animals the immunological memory formation and/or its realization was destroyed to a greater extent and was restored more slowly than the capacity to the primary response.


Plasmapheresis in the treatment of thrombotic thrombocytopenic purpura.

Two patients with thrombotic thrombocytopenic purpura (TTP) have recovered completely after intensive plasmapheresis. The mechanisms responsible for the improvement in these instances are most likely related to the removal of an inciting or damaging agent. The possibility that this agent may be an immune complex is discussed. Plasmapheresis appears to be useful therapy for some patients with this syndrome.


Interactive effect of chromium compounds and a fungal parasite on carp eggs.

A fungal parasite (Saprolegniales) of carp eggs was exposed to hexavalent and trivalent chromium compounds at concentrations identical to those used in a study of the effects of the same compounds on carp eggs hatchability. Comparison of data from both experiments revealed that increased egg mortality and increased fungal growth were cooupled at low concentrations of the compounds. A mechanism for the interaction of a chemical stress and a biological stress on carp egg survival in natural systems is presented.


[Prognostic significance of histological and cytological parameters in biopsies].

In cancer and precancerous conditions, cytologic and histologic investigation of biopsies can serve to establish prognosis as well as for diagnosis. All prognostic factors must be taken into account in modifying a particular tumor therapy. Additional determination of biochemical parameters on the basis of biopsy material makes it possible to characterize the biology of tumor cells with greater precision. Transplant of tumor fragments into nude mice offers a further means of investigating tumor cell properties more closely in vivo.


Sex differences in the distribution of cytoplasmic oestrogen receptors in rat brain and pituitary: effects of gonadectomy and neonatal androgen treatment.

High affinity binding of 17 beta-oestradiol was measured in cytosols of hypothalamus, amygdaloid region, pituitary and (in females) uterus of adult male and female rats. There were no differences between intact or gonadectomised male and female animals in any of the tissues in the equilibrium dissociation constants (Kd). The number of available binding sites (n) in brain and pituitary in intact females at metoestrus is higher than at proestrus but only in hypothalamus is n greater than in ovariectomised animals. Binding sites in male hypothalamus, amygdala and pituitary are significantly less than in metoestrous females; the sex difference is seen also in gonadectomised rats but is significant only in pituitary. In all female animals the highest concentration of binding sites in the hypothalamus is in the anterior part and the lowest in the posterior part. The distribution of binding sites and Kd values in adult females treated neonatally with testosterone propionate were not different from those of intact proestrous rats. In intact males the highest level of n was in mid-hypothalamus; after gonadectomy the pattern reverted to that in females. It is suggest that these results support the concept that testicular androgen is conversted in brain to a substance with affinity for cytosol oestrogen receptor.


Mitochondrial DNA synthesis in mouse L cells temperature sensitive in nuclear DNA replication.

Temperature-sensitive (ts) A 1S9 mouse L cells continue to synthesize double-stranded covalently closed mitochondrial (mt) DNA at a temperature (38.5 degrees C) which is nonpermissive for chromosomal DNA replication. The amount of mt DNA made appears to be quantitatively linked to nuclear DNA synthesis. Nuclear DNA replication proceeds normally for 6-8 h after the cells are shifted to 38.5 degrees C, and then declines to reach a minimum at 20-24 h. The level of mt DNA synthesis remains high during this period and decreases once the ts lesion has been established.


Kinetics of the hydrolysis of cellulose by beta-1,4-glucan cellobiohydrolase of Trichoderma viride.

The cellulolytic enzyme beta-1,4-glucan cellobiohydrolase (CBH) has been isolated from the crude mixture of cellulase enzymes of Trichoderma viride by gel filtration and ion-exchange methods, and some aspects of its kinetic behaviour have been examined. Studies of the initial rates of the CBH-catalyzed production of cellobiose from fibrous alpha-cellulose show that (i) the dissociation constant for cellobiose competitive product inhibition of the reaction is Ki = (1.13 +/- 0.37) X 10(-3) M, (ii) the adsorption of CBH on fibrous alpha-cellulose and its subsequent reaction conform to kinetic equations developed in conjunction with the Langmuir adsorption isotherm, (iii) the rate-pH curve has a maximum at pH 5.2 and decreases at higher and lower pH values, exhibiting enzyme pK values of 3.8 and 6.5, and (iv) the energy of activation of the overall reaction between 5 and 60 degrees C is 5.3 +/- 0.3 kcal mol-1 at pH 5.2. Studies of the time course of the reaction over extended periods of time up to 40% hydrolysis of the cellulose show that (v) the data fit better to a competitive product inhibition model than to models of anticompetitive product inhibition or noncompetitive product inhibition.


Factors in the survival of stored corneas.

In studying the survival of stored corneas we wished to know if excising the cornea was the most critical step or if one storage medium was superior to another. Rabbit corneas were excised and stored in various media at 4 degrees C or 23 degrees C for four days. The storage media included M-K medium, balanced salt solution (BSS) and 10(-8)M concentration of hydrocortisone in M-K medium and BSS. Comparisons were made by measuring acid phosphatase as an index of autolysis. Results showed a reduced amount of autolysis at 4 degrees C as compared to 23 degrees C. No statistical difference between M-K medium and BSS was seen but steroid decreased autolysis. M-K medium plus hydrocortisone seemed to be the best of the solutions studied, but removal of the cornea seemed to be the most important factor in its survival.


[Role of photocoagulation in central serous choroidopathy].

We reviewed thirty-two cases of central serous choroidopathy and retained twenty-five for a twelve-month study of the natural history of this disease. The main conclusions are that the disease is benign and self-limited; it sometimes leaves sequelae such as subnormal visual acuity because of faveolar leak through Bruch's membrane; photocoagulation is useful only in a minority of cases.


Clinical studies of cancer etiology.

Clues to causes of cancer in man can be identified through clinical observation of patients. Alert practitioners have, for example, recently discovered carcinogenic effects of occupational exposure to vinyl chloride and of diethylstilbestrol therapy during gestation. Clinical case studies have also clarified the role of host susceptibility in development of cancer. In most instances, validity of initial clinical observations was established by more detailed epidemiologic and laboratory studies. Clinicians can contribute to carcinogenesis studies by being alert to causes of cancer in their patients, and by referring exceptional observations to clinical epidemiologists and basic scientists for further study. Knowledge of risk factors can be applied to cancer prevention and early detection.


Antimitotic and antitubulin activity of the tumor inhibitor steganacin.

Steganacin, a newly isolated tumor inhibitor, completely inhibits cleavage in sea urchin eggs at 3 X 10(-7) M by preventing the formation of the mitotic apparatus. Steganacin inhibits the polymerization of tubulin in vitro and also causes a slow depolymerization of preformed microtubules. Optical ultracentrifuge studies of steganacin-treated tubulin show a small reduction in 20 S and 30 S peaks at 0 degree. In electron microscope studies the ring structure of tubulin is seen at 0 degree but disappears if the temperature of tubulin incubated with steganacin is raised to 37 degrees. Steganacin inhibits the binding of colchicine to tubulin and thus resembles podophyllotoxin, which also competitively inhibits colchicine binding. Steganacin had a trimethoxybenzene ring and probably interacts with that portion of the colchicine-binding site that recognizes the trimethoxybenzene ring of colchicine.


Partial synchronization of L1210 cells by 5-fluorouracil and its use in drug combinations.

When L1210 cells growing logarithmically were exposed for 8 hr to a nonlethal dose of 5-fluorouracil (FU) (0.25 microgram/ml), the percentage of cells in S phase increased from 74.9% in the asynchronous culture to 93% in the FU-treated culture. This resulted in increased cell-kill by S-phase-specific inhibitors [1-beta-D-arabinofuranosylcytosine (ara-C), 5-hydroxy-2-formylpyridinethiosemicarbazone] when they were added to a culture partially synchronized by pretreatment with FU. For example, 2 hr exposure to ara-C alone or ara-C plus FU (added simultaneously to asynchronous culture) gave 28.8 and 25.8% survival, respectively, compared to 6.8% survival when ara-C was added for 2 hr to the partially synchronized culture. Eight to 12 hr after FU removal, the culture became asynchronous, such that ara-C addition at this time did not result in increased cell-kill. Cultures pretreated with FU were also highly sensitive to vincristine and Adriamycin. Adriamycin acted synergistically with FU (after 8 hr pretreatment) in killing L1210 cells.


Chromosome analysis and alkaline phosphatase of C41, a cell line of human cervical origin distinct from HeLa.

The C41 cell line, which was derived from a human squamous carcinoma of the uterine cervix, has been characterized by analysis of quinacrine-banded metaphase chromosomes and study of alkaline phosphatase. C41 cells have a distinctive karyotype. They are hypodiploid, with a highly characteristic series of marker chromosomes, most of them derived by translocation or deletion. They contain no HeLa cell marker chromosomes, and the cell line shows no evidence of HeLa cell contamination. Nevertheless, the C41 and the HeLa cell line, both derived from cervix cancer, although of a different histological type, produce similar alkaline phosphatases. The enzyme is heat stable (placental type), is inhibited by L-phenylalanine, and responds to the inducing effects of prednisolone and/or hyperosmolality.


Effects of promoters on DNA synthesis in C3H/10T1/2 mouse fibroblasts.

The synthesis of DNA has been studied by autoradiography and by measurements of tritiated thymidine ([3H]TdR) incorporation in cultured C3H/10T1/2 mouse embryo fibroblasts. The cells were first treated with 3-methylcholanthrene as an initiator and then with promoters according to schedules that produce oncogenic transformation. The levels of 3-methylcholanthrene used did not affect the growth or [3H]TdR incorporation of the cells. Treatment during the log phase of growth with 12-O-tetradecanoyl-phorbol-13-acetate, phorbol didecanoate, or 4alpha-phorbol didecanoate produced a transient inhibition of [3H]TdR incorporation with the maximum at 12 hr after treatment. This resulted in a temporary delay of growth followed by recovery of the normal cell-doubling time. Phorbol did not produce these effects, suggesting that the inhibition of DNA synthesis is associated with the process of promotion. Although treatment of the cells with 12-O-tetradecanoyl-phorbol-13-acetate during stationary phase resulted in a 2- to 3-fold stimulation of [3H]TdR incorporation, multiple treatments spanning log and stationary phases were found to be necessary for promotion.


DNA repair in V-79 cells treated with combinations of ultraviolet radiation and N-acetoxy-2-acetylaminofluorene.

Earlier experiments on human cells showed that N-acetoxy-2-acetylaminofluorene mimics ultraviolet radiation in biological and repair characteristics and that the amount of repair from a combined treatment was additive. Chinese hamster V-79 cells are less proficient than human cells in excision repair of pyrimidine dimers resulting from irradiation. We therefore investigated the combined effects of both agents on repair in V-79 cells to see whether they follow the same pattern as in human cells. They did not. Measurements of unscheduled DNA synthesis and the photolysis of DNA repaired in the presence of bromodeoxyuridine gave information about repair due to both agents, and the use of an endonuclease in an extract of Micrococcus luteus allowed us to measure repair of only ultraviolet damage in the presence of N-acetoxy-2-acetylaminofluorene damage. Each technique indicated that the amount of repair from a combined treatment was less than additive and in some cases less than that due to either agent. We conclude that V-79 cells are different from human fibroblasts in the excision repair of both ultraviolet and N-acetoxy-2-acetylaminofluorene damage and suggest that both kinds of damages inhibit repair of damage due to the other agent.


Effects of ethidium and isometamidium on adenosine triphosphate catabolism and purine nucleotide synthesis in Ehrlich ascites tumor cells in vitro.

Ethidium and isometamidium induce the breakdown of intracellular adenosine triphosphate in Ehrlich ascites tumor cells incubated in vitro. Ethidium induces appreciable adenosine triphosphate breakdown only when cells are incubated without glucose, whereas isometamidium produces this effect both in the presence and absence of glucose. In cells treated with isometamidium, purine nucleoside monophosphates accumulate, whereas these are mostly dephosphorylated when ethidium is used. Both ethidium and isometamidium inhibit purine nucleotide synthesis and incorporation of precursors into nucleic acids, although the magnitudes of these effects varied with the precursor used. Isometamidium inhibited the conversion of inosinate to adenine and guanine nucleotides, and both compounds partially inhibited the accumulation of phosphoribosyl pyrophosphate.


Synthesis of heterogeneous nuclear RNA in full-grown oocytes of Xenopus laevis (Daudin).

At various times following injection of either 3H-GTP or 32PO4 into full-grown (stage 6) Xenopus laevis oocytes, RNA has been extracted and fractionated on polyacrylamide gels. Based on size, base composition and incorporation data, we have defined  the kinetics of synthesis and accumulation of ribosomal RNA (40S, 28S, 18S), heterogeneous RNA of high molecular weight (greater than 40S) and heterogeneous RNA migrating with molecular weights of from 4S to 40S. Nuclear isolations have been performed to determine the cellular distribution of these classes of RNA as a function of time. Evidence is presented which shows that stage 6 oocytes synthesize RNA which by virtue of its size, base composition, rapid turnover and nuclear location is equivalent to the heterogeneous nuclear RNA observed in somatic cells. In addition, the data suggest synthesis of a class of nuclear RNA with a half-life of several hours. A small fraction (5%) of the nuclear RNA is stable, enters the cytoplasm and may represent RNA  added to the stockpile of maternal transcripts known to be present in stage 6 oocytes.


The dynamics of maternal poly(A)-containing mRNA in fertilized sea urchin eggs.

Cytoplasmic polyadenylated RNA with the characteristics of sequestered mRNA exists in the unfertilized sea urchin egg. Following egg activation, the amount of poly(A) doubles, but total RNA content stays constant. Chromatography of the RNA on poly(U)-Sepharose shows that the amount of RNA that bears a poly(A) tract increases slightly (approximately 20-30%) during the 2 hr after fertilization. When a cDNA transcript of the poly(A)+ mRNA from 2 hr zygotes is reacted against poly(A)+ RNA from either eggs or zygotes, the kinetics of reassociation of the two preparations seem identical; hence the RNA sequences bearing poly(A) are the same in eggs and zygotes. Measurement of the length of the poly(A) tract in eggs and zygotes shows an increase in number average length from about 45 bases to 60 bases. Measurement of tract length of poly(A) in two cell zygotes by adenosine/AMP ratios of radioactive RNA shows that the poly(A) tract of the zygote is solely accounted for radioactive RNA, indicating extensive turnover of the poly(A). It is concluded that the poly(A) tract in these cells is subject to both lengthening and shortening, with the former predominating in this instance. the increase in poly(A) does not involve polyadenylation of different sequences, but is due to an increase in the number of polyadenylated sequences and the length of the poly(A) tracts that they bear.


Subcellular compartmentation of albumin and globin made in oocytes under the direction of injected messenger RNA.

The Xenopus oocyte can be used to study the nature and specificity of the translational and post-translational systems of a normal living cell. We describe experiments combining messenger RNA microinjection and subcellular fractionation. Total Xenopus liver RNA directs the synthesis of albumin and vitellogenin contained within membrane vesicles; similarly, guinea pig mammary gland mRNA codes for membrane-bound protease-resistant milk proteins. In contrast, iodinated albumin protein injected into oocytes remains in the supernatant fraction, as does globin made on mRNA. The information encoded in the albumin messenger is therefore sufficient to specify synthesis of a membrane-bound product; moreoever, this highly efficient coupled translation-processing system may be of use in the study of the transfer of newly made proteins across membranes. A significant proportion (up to 20%) of newly made oocyte proteins enter a light membrane fraction, and many remain there. We speculate that these vesicles represent part of an important storage system.


DNA replication during muscle cell differentiation: identification of multiple DNA-dependent DNA polymerases.

DNA-dependent DNA polymerases have been studied during chick embryo muscle differentiation in vitro. The total activity, extracted at both low and high ionic strengths, does not change throughout the differentiative process, although DNA synthesis stops at the moment of fusion. Analyses by glycerol gradient centrifugation of the extracts at low and high ionic strengths show two major DNA polymerase forms, one sedimenting at 7.5 S and another at 3-4 S. Both enzymes are present in similar amounts in duplicating myoblasts and in post-mitotic myotubes. These data suggest that the arrest of DNA synthesis which accompanies myoblast differentiation is not dependent on the disappearance or decrease of the major DNA polymerase activities described.


Angled views in cineangiocardiography of congenital heart disease.

Simple and compound sagittally angled views of the heart obtained cineangiocardiographically with a parallelographic U-arm device were evaluated clinically in 132 patients with congenital heart disease. The principle advantage of the apparatus was the case with which axial and oblique projections were obtained without repositioning of the patient, although rotation and angulation of the image intensifier-X-ray tube unit resulted in some increase in scattered radiation. Certain angled views, particularly the 40 degree cranial and the compound 25 degrees cranial/70 degrees left oblique projections, were better than standard frontal and lateral views for visualization of pulmonary arteries in the mediastinum, in ventricular septal defects and anomalies of the left ventricular outflow tract.


Surgical treatment of idiopathic hypertrophic subaortic stenosis (IHSS). Postoperative results in 30 patients following ventricular septal myotomy and myectomy (Morrow procedure).

Since 1971 we have employed the Morrow procedure in 30 patients with idiopathic hypertrophic subaortic stenosis (IHSS). All manifested obstruction to left ventricular outflow either at rest or with provocation (Valsalva) and 17 had moderate or severe mitral regurgitation. There were no operative deaths. Obstruction was abolished in all patients except one in whom a trivial 15-mm pressure gradient persisted, and all patients with moderate or severe mitral regurgitation evidenced marked amelioration of the mitral regurgitation. Distinct symptomatic improvement has been experienced by all patients, and 20 are entirely asymptomatic postoperatively. No patient has experienced syncope postoperatively although 15 experienced syncope before operation. In 12 patients the electrocardiogram recorded postoperatively was essentially unchanged from the preoperative record. In 16 patients a left anterior hemiblock was apparent postoperatively. In one patient a complete left bundle branch block appeared postoperatively and one patient demonstrated Wolff-Parkinson-White syndrome. One patient died suddenly and unexpectedly 2 years following operation. This patient continued to have palpitations after operation although all other symptoms as well as the left ventricular outflow obstruction were abolished by operation. It is suggested that propranalol administration be continued postoperatively in those patients experiencing palpitations or manifesting arrhythmias. Asymmetrical septal hypertrophy dislocates the cardiac apex and papillary muscles anteriosuperiorly producing abnormal systolic anterior mitral leaflet mition. The Morrow procedure restores more normal ventricular geometry and thereby eliminates the pathophysiological mechanism of obstruction and mitral regurgitation. The myotomies and myectomy should be extended far inferior through the entire width of the septum onto the free lateral ventricular wall. The procedure can be accomplished easily and safely through a transaortic approach. Since the operative risk at present appears to be negligible, prosthetic material is not required, and the risk of significant injury to the conduction tissue is small, we feel the current status of surgery for IHSS is such that the Morrow procedure should be performed earlier in the course of patients with IHSS manifesting obstruction or mitral regurgitation.


The radioimmunoassay of human lactalbumin.

A radioimmunoassay has been developed for the measurement of human lactalbumin, using the protein isolated in pure form from breast milk. Under disequilibrium conditions with second antibody separation the system will detect 20 pg of lactalbumin. Lactalbumin is detectable in the serum of approximately a third of normal male subjects; among women theproportion having measurable serum levels is age-related. Very much higher levels are present in the serum of lactating women.


Clinical picture of orf in Northern California.

Eighteen cases of orf are reported, six with multiple lesions and twelve with only one. Iridoform bullae were seen in every case, and untraumatized lesions healed without scar. The fingers and hands were involved in fourteen instances, the remainder occurring on the forearms, nose, forehead, and eyelid. Lack of systemic symptoms was usual, although several patients had regional adenopathy, malaise, and fever. Treatment was purely symptomatic, and x-ray therapy was not effective.


[Clinical aspects, radiology and therapy of angiosarcoma of the breast gland].

A case of angiosarcoma of the breast is described. The problems of clinical appearance, roentgenology and therapy are discussed. The case described is similar to those seen in the literature, of which 44 cases are known in the literature. Problems of diagnosis even during surgery are evident.


Laser application to the angle structures in rabbits, cats, and in human glaucomatous eyes.

Argon laser lesions were induced to the structures of the anterior chamber angle in rabbits and cats and in a pre-enucleated infant's eye. Histopathological changes at different follow-up periods were analyzed. The argon laser beam was also applied to 28 uncontrolled glaucomatous eyes. Twenty-five eyes responded by an abrupt hypotensive effect, but in 21 of them, elevated pressure recurred after 20 to 40 days. Eight eyes maintained low intraocular pressure throughout the 12 to 20 month follow-up period. Of these, 3 were eyes of babies with congenital glaucoma, and 3 had simple glaucoma following an unsuccessful filtering operation. Seven eyes were medically controlled postoperatively and 13 eyes were failures which had to undergo further surgery.


Drug-seeking behavior in the dog: lack of effect of prior passive dependence on morphine.

Twelve dogs were made dependent on morphine (20 mg/kg per 24 hours) by intravenous passive administration of the drug. Six were gradually withdrawn and six were withdrawn abruptly. Subsequent tests for self-administration of morphine began within 8 to 18 weeks after morphine was last administered passively. Morphine was available for self-administration at several unit dose levels for 8 weeks. In comparison with control dogs administered with saline, there was no evidence that prior dependence on morphine influences subsequent self-administration of morphine, or that morphine is a primary reinforcer for dogs. Possible implications of these and related observations are discussed.


L-Dopa inhibits prolactin secretion in proestrous rats.

Female rats received an ip injection of L-dopa on the afternoon of proestrus. L-Dopa reduced serum prolactin concentrations within 1 h, whether administered just prior to, or during, the normal surge in serum hormone level. This inhibition lasted for 2-3 h, after which serum prolactin concentrations rose substantially. Pretreatment of proestrous rats with MK-486, a peripheral inhibitor of aromatic-L-amino acid decarboxylase, did not block the effect of L-dopa on serum prolactin levels. In fact, MK-486 pretreatment appeared to prolong the effectiveness of L-dopa. Pretreatment with RO4-4602 at a dose sufficient to block central decarboxylase activity, however, did prevent dopa from inhibiting the proestrous surge in serum prolactin. These data are consistent with a role for dopamine in the control of prolactin secretion and suggest that the mechanism of action of L-dopa apparently does not require peripheral decarboxylation.


Effect of melatonin administered during the night on reproductive function in the Syrian hamster.

Syrian hamsters were maintained on a long photoperiod (lights on 0600-2000 h) and injected daily with melatonin (10 microgram per injection) for 6-7 weeks. Females injected at 2000 h or 0400 h ceased estrous cyclicity, and males treated similarly at these times had reduced testis weights by the sixth week of treatment. Daily administration of the same dosage of melatonin at 2400 and 0100 h failed to interrupt estrous cyclicity or reduce testis weight. These observations suggest that exogenous melatonin administered at 2000 h or 0400 h may be temporally adding to endogenous circulating melatonin, inducing gonadal quiescence.


Two immunologically distinct human acidic beta-galactosidase A isozymes.

Two acidic beta-galactosidase isozymes (designated A1 and A2) were separated by isoelectric focusing from beta-galactosidase A of human liver. Kinetic studies with 4-methylumbelliferyl-beta-D-galactopyranoside substrate revealed similar parameters for both. The Km value was 0.32 mmol/1 for A1 and 0.30 mmol/1 for A2 and Vmax values of 59.3 and mumol min-1 mg-1, respectively. The pH optimum was 4.2 for beta-galactosidase A1 and 4.5 for the A2 form. The A1 enzyme form was shown to be more heat labile than the A2. Significant differences were observed with antibody preparations against the two enzyme forms. Using the anti-A1 antibodies two precipitin arcs with residual enzymatic activity were obtained by immunoelectrophoresis of beta-galactosidase A whereas only one with anti-A2 antibodies. Anti-A1 precipated 85% of the original activity present in beta-galactosidase A and only 56% could be precipated by anti-A2. The possibility of common structural components is suggested.


The determination of molecular-motion parameters from proton-relaxation-enhancement measurements in a number of Gd(III) - antibody-fragment complexes. A comparative study.

Longitudinal and transverse proton relaxation rates for water in the hydration spheres of Gd(III) bound to the non-immune rabbit IgG fragments Fc (C-terminal half of heavy-chain dimer), pFc' (C-terminal quarter of heavy-chain dimer) and Fab (N-terminal half of heavy and light chain) have been measured at a number of frequencies and temperatures using pulsed nuclear magnetic resonance spectrometry. For the fragments Fc and pFc', a full computer analysis showed that the results could be fitted by parameters of similar magnitude to those found previously for IgG. In contrast to the results for the other complexes the Fab -Gd(III) complex showed no slow exchange contribution to the relaxation rates. Under these circumstances it was found possible to obtain an accurate value for the hydration number (q) from measurements of the longitudinal and transverse relaxation rates at a chosen frequency such that the product of the nuclear Larmor frequency (omega1) and the correlation time for the dipolar relaxation processes (tauc) was approximately unity. Water-proton relaxation rates were also determined for the complex of Gd(III) with the Fv fragment of the mouse myeloma protein MOPC 315. A computer analysis of the results revealed a slow exchange contribution to the rates and this gave errors in the variable parameters similar to those observed previously for IgG, Fc and pFc'. The conclusions drawn from the different systems are discussed in terms of the present state of application of the proton relaxation enhancement technique in biology.


The organisation of genes for transfer RNA and ribosomal RNA in amoebae and plasmodia of Physarum polycephalum.

1. Using hybridisation techniques nuclei from both amoebae and plasmodia of Physarum polycephalum were found to contain 275 genes each coding for 5.8-S, 19-S and 26-S rRNA, 685 genes for 5-S rRNA and 1050 genes for tRNA. 2. Hybridisation of these RNA species to both amoebal and plasmodial DNA fractionated on CsCl gradients reveal that the 5.8-S, 19-S and 26-S rRNA genes are located at a satellite position (formula: see text) with respect to the main band of DNA, whereas 4-S RNA genes are located exclusively in the main band of DNA (formula: see text). 3. This result was confirmed by demonstrating that only the 5.8-S, 19-S and 26-S rRNA species hybridise to purified plasmodial ribosomal DNA. 4. The 19-S and 26-S rRNA genes of amoebae are located on extrachromosomal DNA molecules of a discrete size (Mr = 38 X 10(6)) with identical properties to plasmodial ribosomal DNA.


Tuberculosis and infertility in men.

In 50 patients, all under 40, a diagnosis of urinary tuberculosis and a spermiogram have been made in order to see if there was any cytomorphological or biochemical alteration. We have divided them into two groups: the first one including the patients showing clinical alterations of the genital structures, and the second one including those for whom it had not been possible to demonstrate any clinical symptomatology, e.g. at the external genitalia. The modification of the spermiogram in accordance with the evolution of the urinary tuberculosis lesion and the specific treatment employed has also been evaluated. All patients with clinical alterations in the genital structures showed alterations in the cytomorphological and/or in the biochemical study. 75% of the patients without genital lesions showed oligoasthenozoospermia. We have not been able to observe any improvement whatsoever in the spermiogram in spite of the treatment established.


Immunological evaluation in patients with urological cancers.

The immunological competence of 100 patients bearing urological cancers was evaluated by means of skin tests and serum immunoelectrophoresis. Cell-mediated immunodepressions were observed in 93.3% of prostatic cancers, 68.6% of bladder cancers and 38.4% of renal cancers. The immunoglobulin values increased in 80% of prostatic cancers, 31.3% of bladder cancers and in 23% of renal cancers. IgA was elevated in almost all the prostatic growths. In bladder and renal cancers a clear correlation between immunodepression and tumour stage was found. Treatment significantly improved the skin test reactivity in the evaluated patients. Metastases and recurrences occurred more frequently in immunodepressed subjects.


The kinetics of ethanol absorption and elimination in twins and supplementary repetitive experiments in singleton subjects.

The influence of genetic and environmental factors on the metabolism of a single oral dose of ethanol 1.2 ml per kg body weight was analysed in 19 identical and 21 fraternal healthy, adult, un-selected male twin pairs. The heritability values of the rates of absorption, degradation and elimination of ethanol were 0.57, 0.41 and 0.46 respectively. Environmental factors, such as daily alcohol intake and smoking, increased the rate of elimination of blood alcohol. Intrasubject variation in ethanol metabolism was studied by repeated tests in 11 male volunteers at intervals of at least 2 months, under the same conditions as in the twins; the coefficients of variation for parameters of metabolism was about 8%. The results demonstrate both genetic control of ethanol absorption, degradation and elimination and the appreciable influence of environmental factors. The almost total genetic control of ethanol metabolism postulated by Vesell et al. (1971) could not be confirmed.


Brain serotonin turnover and morphine tolerance-dependence induced by multiple injections in the rat.

Tolerance to and physical dependence on morphine in the rat was induced by injecting increasing doses of morphine sulfate (M.S.) administered i.p. twice daily for 14 days. The last dose of M.S. was 200 mg/kg. This procedure produced a 4-fold tolerance to morphine as evidence by the increased dose of morphine required to produce analgesia. The degree of dependence was quantified by determining the naloxone ED50 for the stereotyped withdrawal jumping response. Body weight loss and hypothermic responses during abrupt and naloxone-induced withdrawal were also measured. The degree of tolerance and dependence produced by multiple injection procedure was comparable to that produced by 2 pellets containing 75 mg of morphine base implanted for 3 days. The level and turnover of brain serotonin, determined 6 or 12 h after the last morphine sulfate injection did not differ significantly from that of saline injected control animals. These data indicate that multiple injection technique produces a mild degree of tolerance to, and physical dependence on, morphine which was not related to changes in brain serotonin level or turnover.


Comparative effects of hallucinogenic drugs on rotational behavior in rats with unilateral 6-hydroxydopamine lesions.

The dopaminergic actions of various hallucinogenic drugs were assessed by examining their effects on turning behavior in rats with unilateral 6-hydroxydopamine lesions of the nigro-striatal pathway. LSD (0.1 and 0.2 mg/kg) produced strong contralateral turning, indicating that it is a potent dopamine receptor agonist, while BOL (5 mg/kg), a non-hallucinogenic congener of LSD, was found to be a weak dopamine receptor agonist. STP (2 and 5 mg/kg) and mescaline (50 and 100 mg/kg) produced significant ipsilateral turning, indicating that these compounds have a moderate dopamine-releasing action. DMT (10 and 20 mg/kg) and 5-M-DMT (0.75 and 1.25 mg/kg) produced weak ipsilateral turning, which was not significantly different from that produced by the nonhallucinogenic compounds tryptamine (40 mg/kg) or scopolamine (0.25 mg/kg). Psilocin (1-20 mg/kg) produced no significant turning in either direction. These data, in conjunction with previous studies, indicate that while inactivation of the brain serotonin system may be a necessary and sufficient condition for hallucinogenesis, the ability to activate dopamine receptors may be an important factor in determining the potency of hallucinogens.


[Comparative studies on bone growth in experimental animals. 1. Bone growth and ossification in mice (author's transl)].

Comparative studies are needed on the bilogical stage of the bone growth in experimental animals and human to extrapolate the data from animals to for estimating the risk of bone seeking redionuclides in man. The growth of bone length and the appearance and fusion of the secondary ossification centers of seven long bones were compared between both species. The stage of ossification was determined radiographically. The results showed that the ossifying process of the secondary ossification centers could be divided into the following three categories in mice; (1) acute ossification type, (2) remaining metastable stage type which is specific in mice and generally at the proximal end of bone, and (3) delayed ossification type. Compared with the average life span, differences in ossification between both species were that the appearance of secondary ossification centers was observed within concentrated period in mice and not in man, and that the fusion of these was observed with long duration in mice and not in man. From 17 weeks of age, the metastable status of the bone appeared in mice without complete fusion even at 27 weeks of age, for the experiment with bone seeking radionuclides, mice of age 17 weeks or more should be used consequently.


[An outbreak of "wet tail" in hamsters due to Tyzzer's organism (author's transl)].

In a colony of Golden hamsters, many animals were found to show signs of "wet tail", and spontaneous deaths occurred during two months. With this outbreak of wet tail and death, etiologic and pathologic studies were made on 11 cases supplied from the colony. Gross pathologic examination disclosed hemorrhagic enteritis in 10 and necrotic foci in the liver in 5 out of 10 cases examined. A number of bacilli morphologically suggestive of Tyzzer's organisms were detected on smears of cut surfaces of the affected intestines and livers. After inoculation with homogenate of the intestines from the moribund cases into hamsters by oral route, the same clinical manifestations as observed in spontaneous cases were reproduced, and the inoculated animals died in 10 to 14 days with numerous Tyzzer's organisms in their intestines and livers. When apparently healthy hamsters were introduced into contaminated cages with soiled bedding which were allowed to stand at room temperature for about 2 months, most of them developed signs of wet tail and died in 5 to 19 days having liver necrosis characteristic of Tyzzer's disease.


[A simple operation method on removal of olfactory bulbs of small rodents (author's transl)].

A simple operation method was described for removing the olfactory bulbs of small rodents. The procedure was as follows: 1. Animals were anesthetized by intraperitoneal injection of pentobarbital sodium. 2. The hair of the head was clipped (Fig. 9). 3. The scalp was cut along the midline between the eyes (Fig. 10). 4. A hole was drilled in the skull overlying the olfactory bulbs (Fig, 14, 15). 5. The olfactory bulbs were aspirated with a glass pipette (Fig. 18). 6. After removing the bulbs by suction, the skin was closed by a single suture (Fig. 21).


[A new microtiter plate agglutination method for Brucella canis antibodies (author's transl)].

A micro-agglutination test method for detecting antibodies to Brucella canis was developed. Heat-killed Brucella canis antigens were diluted to an optical density of 0.8 at 420 nm using a spectrophotometer. A volume of 0.025 ml of the antigen was incubated with the same volume of serially diluted sera for 18 to 24 hr at 37 C. Titers of selected dog sera obtained by the present micro-test method were well correlated with those obtained by the classical tube test with satisfactory reproducibility. The micro-test method is more advantageous for screening the antibodies of dog sera because the test can be performed with: (1) smaller volume of the test sera and the antigen (2) shorter period for incubation, and (3) lesser labor.


Aerophagia induced by the nasal obstruction on experimental animals.

The excessive accumulation of gas in the gastrointestinal tracts was invariably induced on experimental animals (mice, rats, guinea pigs, hamsters and rabbits) by simply obstructing nasal passages. The analysis of the gas showed the almost identical composition to the ambient air or flutus which was largely due to swallowed air. Also the numerous small foams were found on and underneath the epithelial lining of small intestine. The pathological evaluation was done both macroscopically and microscopically. Dying animals after nasal obstruction showed hemorrhagic and necrotic changes in the jejunum and ileum. This observation may cast some light to the pathogenesis of necrotizing enterocolitis in human neonatal.


Spontaneous lesions detected in the beagles used in toxicity studies.

Four hundred and twenty beagles 1 to 4 years of age were examined for spontaneous pathological changes. In the liver were seen fine granular gallstones (87.5%), granulomas (59.9%), intranuclear hyaline inclusions (52.6%), lipofuscin deposition (50%) and eosinophilic inclusions (3.7%) in the cytoplasm of hepatocytes and parenchymal necrosis at the base of the ligaments (7.3%). Intranuclear hyaline inclusions and lipofuscin deposition were more frequently seen in dogs after being subjected to tests for more than 13 weeks. Besides, hyperplasia of follicular epithelial cells of the thyroid gland (46.3%), Gandy-Gamna like bodies of the splenic capsule (43.1%), small cysts in the anterior lobe of the hypophysis (35.1%) and prostatitis (13,7%) were observed. Lesions due to migration of Toxocara canis larvae were detected in the liver, kidney, lung and other organs from 33.1% of cases examined.


Diurnal variation of visual field size in patients with postretinal lesions.

Perimetry at various times of day in patients with large visual field defects due to postretinal lesions showed significant variations of visual field size. The largest visual fields were observed at noon, the smallest in the evening. Such systematic variations were observed only in patients who showed a gradual increase of increment threshold between the intact parts of the visual field and the scotoma. In two patients who showed an abrupt transition between intact and blind areas of the visual field, no obvious diurnal variation was observed. It is suggested that an endogenous modulation of neuronal sensitivity coupled to a hypothetical circadian oscillator is the basis of the diurnal variation.


Time course of ovum transport in guinea pigs.

The time course of ovum transport was determined in cycling guinea pigs. The occurrence of ovulation was determined in assessing changes in vaginal cytology. The day of the postovulatory leukocyte influx was considered day 1 of the cycle. Ovum transport in guinea pigs is characterized by a relatively long sojourn in the ampulla, followed by rapid transport through the isthmus. This pattern is similar to that seen in women and subhuman primates and different from the pattern in rabbits.


[Effect of thyroxine and insulin on nitrogen metabolism in the gastrointestinal system and mammary glands of lactating cows].

In 9 cows giving 6-10 kg of milk daily and fed with 4 kg of combined food, administration of 50 mg of thyroxin increased extraction of some serum proteins from the blood by the digestive system (alpha- and beta-globulin factions in particular). Apsorption of amino-nitrogen from the blood by the mammary gland was increased by 3.5 times, and of ammonia by 5 times. Excretion of urine into the blood displayed a three-fold increase. Insulin in dosage 1 unit per 1 kg of body weight increased excretion of serum proteins and urine from the digestive system into the blood. Excretion of gamma-globulins into the lactic vein's blood increased by 3 times, and 0.17% of albumins were exctracted by the mammary gland from the coming blood. Subcutaneous administration of thyroxin and insulin obviously increases the amount of protein, casein and nitrogen substances in the milk.


Mutations causing transformation of sexual phenotype in the nematode Caenorhabditis elegans.

Ten mutations are described that transform genotypic hermaphrodites of the nematode Caenorhabditis elegans into phenotypic males. These fall into three autosomal complementation groups, termed tra-1, tra-2, and tra-3. Two alleles of tra-1 produce almost complete transformation, to a fertile male phenotype; such transformed animals are useful for analyzing sex-linked genes. All alleles of tra-1 and tra-2 are recessive; the one known allele of tra-3 is both recessive and maternal in effect. Where tested, both XX and XXX hermaphrodites are transformed into males, but XO males (true males) are unaffected by these mutations. It is suggested that these genes are actually involved in hermaphrodite development and have no role in male development.


Avirulence and decreased virulence in mutants of Emmonsia crescens, a causative agent of adiaspiromycosis.

Three mutants of Emmonsia crescens with altered morphology and pathogenicity for mice are described. The mutant M-1 is avirulent and does not form adiaspores of normal morphology on agar medium at 37 degree C. The mutants M-2 and M-12 have a considerably decreased virulence. The pathogenicity of E. crescens depends on the ability to differentiate adiaspores of perfect morphology. The loss of virulence and the decrease of virulence of mutants seem to be caused by the pleiotrophic effect of mutation of the genetic base, which regulates the differentiation of adiaspores. The conidia of the wild strain of E. crescens killed by UV-irradiation do not induce the formation of granulomas with adiaspores after i.p. inoculation into mice.


[Pharmacological studies of Shikon and Tooki. (2) Pharmacological effects of the pigment components, Shikonin and acetylshikonin].

Pharmacological effects of Shikonin and Acetylshikonin, pigment components of Shikon were studied in mice, rats, guinea pigs and rabbits. The only difference between the chemical structures of the two pigments is that Acetylshikonin has the acetyl radical, while Shikonin does not. Though the activity was slightly different between the two compounds, the pharmacological effect of Shikonin was similar to that of Acetylshikonin. As the two pigments were extracted by ether from Shikon, systemic administration of these pigments showed the same effect as that of ether extract of Shikon as outlined in the first report(1). These compounds had no effect on blood coagulation, but inhibited the anticoagulant effect of heparin in rats. Topical application of both pigments (50 mg of 0.1% ointment) inhibited an increased vascular permeability and acute edema induced by histamine, anti-rat rabbit serum and heat. The activity was similar to that of 0.1% phenylbutazone ointment. On the other hand, these pigments increased proliferation of granuloma tissue in the cotton pellet method and promoted would healing in rats. The results suggest that Shiunko, a main prescription of Shikon, is an effective ointment for cutaneous injuries.


[Pharmacological studies of Shikon and Tooki. (3) Effect of topical application of the ether extracts and Shiunko on inflammatory reactions].

Pharmacological effects of topical application of Shiunko, a main prescription of Shikon (Lithospermum Officinale Linńe var. erythrorhizon Maximowicz), were studied in comparison with that of ether extracts of Shikon and Tooki. Ointment of Shikon ether extract (50 mg/site) inhibited markedly an increased vascular permeability and acute edema induced by histamine, serotonin, bromelain, anti-rat rabbit serum and heat. It also inhibited an increase of local cutaneous temperature induced by phlogistic stimulation (ultraviolet irradiation and heat). On the other hand, it accelerated wound healing in rats. The activity of Shikon ether extract on inflammatory reaction was the most potent in 0.2 approximately 0.1% of ointment concentration and was diminished in higher or lower concentrations. Tooki extract inhibited slightly an acute inflammatory reaction, but had no effect on increase of cutaneous temperature and wound healing in rats. Shiunko contains about 0.2% of Shikon and 0.04% of Tooki extract. The effect of Shiunko was similar to that of Shikon ether extract, but the activity was slightly more potent than that of Shikon. Shiunko inhibited rubor, calor and tumor in acute inflammatory reaction and promoted wound healing. Furthermore, it was reported that pigments of Shikon have antibiotic activity. The results suggest that Shiunko is an effective ointment for cutaneous injuries.


[Effects of morphine on the single unit activity of nucleus dorsal raphe].

Effects of morphine were studied in 36 gallamine triethiodide immobilized adult cats under light N2O anesthesia. Single units were recorded from the nucleus dorsal raphe using a stainless steel microelectrode. Dorsal raphe neurons were divided into two types; one was a clock-like (CL) neuron which was typically slow in rate, rhythmic and stable throughout recording time, and the other was a non-clock like (NCL) neuron which was relatively irregular in pattern as compared with CL neuron. Out of 36 neurons recorded in this experiment, 13 were CL neurons and 23 were NCL neurons. Mean discharge rates (spikes per sec) of CL neuron and NCL neuron were 2.84 (range of 2.01 approximately 3.68) and 4.11 (range of 0.10 approximately 35.09), respectively. None of the 13 CL neurons responded to the nociceptive (pinch and/or brdaykinin) and non-nociceptive (hair bending and/or tapping) stimuli. On the contrary, out of 23 NCL neurons, 13 responded to both nociceptive and non-nociceptive stimuli, 4 were responsive to only non-nociceptive stimuli and 6 were not responsive to these stimuli. Receptive fields of NCL neurons were wide with various somatic modalities. The latency for bradykinin (3 microgram) was 6.46 +/- 0.77 sec. Effects of morphine were examined in 6 CL neurons and 12 NCL neurons. After morphine, no nociceptive neuron responded to nociceptive stimuli, although there was no appreciable change in the responsiveness to non-nociceptive stimuli. Firing frequency and pattern of all CL neurons were unaffected by morphine. Out of 12 NCL neurons, 7 were unaffected in firing frequency and 5 were decreased after morphine.


[Behavioral effects of 5-(o-chlorophenyl)-1-methyl-7-nitro-1,3-dihydro-2H-1,4-benzodiazepin-2-one (ID-690)].

Effects on the central nervous system of ID-690, a new benzodiazepine derivative were investigated and compared mainly with those of diazepam in mice and rats. Locomotor activity of the rat was increased with ID-690, at doses of 2 approximately 5 mg/kg p.o., as with diazepam. ID-690 was approximately 5 times more potent than diazepam in inhibiting fighting behavior of long-term isolated mice. Hyperemotionality induced by either septal lesions or olfactory bulbectomy was also inhibited by ID-690. This effect was almost the same in potency as that of diazepam. ID-690 was only 1/30 as potent as diazepam in preventing maximal electroshock convulsion, while it was approximately 18 times more potent that diazepam in suppressing pentetrazol convulsion in mice. ID-690 was approximately 5 times as potent as diazepam in impairing rotarod performance in mice. The muscle relaxant effect of ID-690 was approximately 10 times as potent as diazepam, as measured with an inclined screen test in mice. Thus the effect of ID-690, as compared with diazepam, was characterized by its relatively potent muscle relaxant and extremely potent anti-pentetrazol convulsant action.


The digestive utilization of the diet in calves experimentally infected with Ostertagia ostertagi.

The experimental infection of calves with 250,000 L larvae of Ostertagia ostertagi per animal did not produce great variations in the digestibility of the diet administered. The infection retarded normal growth the resulted in an unfavourable conversion index.


A 3/4 reciprocal translocation in two unrelated families.

Two cases of reciprocal translocation between chromosomes 3 and 4 found in unrelated families are described. The meiotic consequences of this rearrangement are discussed.


The 48, XXXX/49,XXXXY/49,XXXX,i(Yq) mosaicism in a 3-year-old boy from a twin pregnancy.

A 3-year-old boy from twin pregnancy with the features of marked dystrophia from birth, deficient growth, considerable retardation of physical and mental development, numerous somatic defects, suspected congenital heart disease, and hypoplastic external genitalia, is reported. The 48,XXXX/49,XXXXY/49,XXXX,i(Yq) karyotype was diagnosed. The boy's brother, normally developed, had a 46,XY karyotype. It was found on the basis of serologic findings that the brothers were dizygotic twins.


Uracil phosphoribosyl transferase activity of mycoplasma and infected cell cultures.

Human H.Ep-2 and mouse 3T6 cells infected by Mycoplasma hyorhinis showed an increase in [3H]uracil uptake and a more than 20-fold increase in the activity of uracil phosphoribosyltransferase (UraPRT). Uninfected cell cultures gave background levels of this enzyme activity. A survey of 16 strains of mycoplasma showed 13 to possess UraPRT activity. Rabbit kidney cells (RK13) were infected with eight different strains of four mycoplasma species known to be common cell culture contaminants. Seven of the eight cell cultures showed elevated UraPRT activities four days after infection. This enzyme activity may be of value in monitoring cell cultures for mycoplasma and aid in classification.


Effects of polynucleotides and levamisole on alveolar macrophage morphology and receptor activity.

Incubation of rabbit pulmonary alveolar macrophages in vitro with polyinosinic acid-polycytidylic acid [poly(I-C)] or levamisole results in enhanced immunoglobulin G receptor activity in comparison to untreated cells, Electron microscopy of cells treated with levamisole or poly(I-C) revealed mitochondrial swelling and cytoplasmic vacuolization. The modulation of receptor activity by these agents suggests that their immunopotentiating effects are due to direct simulation of the mononuclear phagocyte system. Lavaged alveolar macrophages have the capacity to change membrane function in vitro, and these cells provide a convenient system for studying agents with potential effects on macrophages.


Value of ultrasound as a presurgical diagnostic tool in Obstetrics and Gynecology.

Although relatively new, diagnostic ultrasound is an important part of the surgeon's armamentarium in preoperative diagnosis and patient preparation. In the field of gynecology, it is possible to differentiate between solid and cystic tumors, measure the extent of tumors in the pelvis and abdomen, define normal structures in relationship to pathologic changes and diagnose and follow patients with malignant disease. In obstetrics, ultrasound has proved effective in the evaluation of the fetus and detection of obstetric problems such as estimation of fetal weight and gestational age, diagnosis of placenta previa, hydatidiform mole and missed and incomplete abortions.


Occurrence of 11-cis-retinal-binding protein restricted to the retina.

A binding protein for retinal (vitamin A aldehyde), the 11-cis-retinal-binding protein, was present in the soluble protein fraction of rat retina but was absent from brain, lung, heart, skeletal muscle, liver, kidney, spleen, small intestine, and testes. The binding protein was also found in human retina but not human liver. The binding protein for retinal from human retina did not bind retinol and was larger than, and readily separable from, the cellular retinol-binding protein which did not bind retinal. It would appear that the occurrence of a soluble binding protein for retinal, unlike the more widely distributed binding proteins for retinol and retinoic acid, may be unique to the retina.


Effects of exogenous follicle-stimulating hormone and luteinizing hormone in bitches.

Ten mixed-breed bitches of unknown ages, weighing 10 to 15 kg, were given a series of subcutaneous injections of follicle-stimulating hormone according to 2 different schedules, for total doses of 35 or 50 mg. Forty-eight hours after the last injection of follicle-stimulating hormone, each bitch was given 15 mg of luteinizing hormone, intravenously. The internal genitalia of 8 of the 10 bitches were recovered 24 hours after the luteinizing hormone injection and were examined histologically. External signs of estrus developed in 7 of 10 bitches in the 2 groups, indicating that estrogen was being secreted by the ovaries. Histologic examination of the ovaries revealed that many follicles had grown to the antral stage, but none had developed to the preovulatory stage, and all were atretic. Ovulation did not occur in any bitch. Endometrial proliferation and vaginal cornification that occurred in 7 of the 10 bitches were further evidence of estrogen production by the ovaries.


Estrous cycle variations of food and water intake in rats in the heat.

Effects of heat and estrous cycle on food and water intake and on the kinetics of water metabolism were studied in Holtzmann rats, 12 adult females per group. The animals were housed individually in metabolic cages, in series, in a controlled-environment room at (24.5 degrees C) followed by two experimental temperatures (29.2 degrees C and 34.0 degrees C). In all cases relative humidity was 50% and the photoperiod 12L:12D (L = 0600--1800 h). Intake of food and water, available ad libitum, were measured and recorded daily for at least five consecutive estrous cycle (about 25 days). Water turnover was measured using tritiated water. The results indicate that the experimental conditions constituted stressful environments for the rats. Their corticosterone levels were significantly (P less than 0.01) elevated and a relative lymphopenia and neutrophilia (P less than 0.01) were observed. Additionally, food intake was decreased and water intake increased, both significantly (P less than 0.01). Body water turnover was increased (P less than 0.01) while body water pool size and the biological half-life for 3H2O were reduced. However, the cyclic variation of food and water intake and its relationhsip to the estrous cycle were unchanged.


Viability of Physarum polycephalum spores and ploidy of plasmodial nuclei.

Amoebae of Physarum polycephalum carrying the mth mating-type allele may differentiate into plasmodia in the absence of mating. Such plasmodia are haploid and, upon sporulation, produce mainly inviable spores. We have asked whether the viable spores arise from meiotic or mitotic divisions. Using a microfluorometric measurement of the deoxyribonucleic acid content of individual nuclei, we found the fraction of viable spores to be correlated with the proportion of rare, diploid nuclei containing in the generally haploid plasmodium. When homozygous diploid plasmodia were created by heat shocking, spore viability increased dramatically. We suggest that viable spores are produced via meiosis in mth plasmodia, that the mth allele has no effect on sporulation per se, and that the normal source of viable haploid spores is a small fraction of diploid nuclei ubiquitous in haploid plasmodia.


Affinity of glucose analogs for alpha-glucan phosphorylases from rabbit muscle and potato tubers.

The action of phosphorylase b from rabbit muscle and potato phosphorylase was inhibited to various extents by several glucose analogs. Like glucose itself, all of the glucosidic oxygen-substituted analogs tested in kinetic experiments showed a nonlinear competitive inhibition for muscle phosphorylase b and a linear competitive one for potato phosphorylase. 5-Thio-D-glucose, one of the ring oxygen-substituted analogs, also inhibited the action of muscle phosphorylase b in the same manner, while the inhibition pattern of 5-amino-D-glucose (nojirimycin) was of a linear noncompetitive type. Since the conformation of 5-amino-D-glucose in aqueous solution is half-chair (Reese et al. (1971) Carbohyd. Res. 18, 381-388), the unusual kinetic behavior of the compound toward muscle phosphorylase b was supposed to be due to its half-chair conformation. In the glucosidic oxygen-substituted analogs, the affinity for both muscle phosphorylase b and potato phosphorylase decreased with decreasing order of magnitude of electronegativity of the glucosidic atom. The strong positive correlation between the affinity and the electronegativity suggests that D-glucose-1-P, the substrate, may bind to phosphorylase with the formation of a hydrogen bond between its glucosidic oxygen and a hydrogen donor of the enzyme.


Intramolecular disulfide linked alphabeta and alphaalpha in oxidized tropomyosin: separation, identification, and process of formation.

The oxidation of rabbit skeletal tropomyosin (TM) by repeated cycles of freezing and melting in 0.3 mM Na bicarbonate was studied by electrophoresis and column chromatography. The oxidized TM showed two bands at ca. 70,000 daltons on sodium dodecyl sulfate (SDS)-polyacrylamide gels. Each band component was separated into disulfide-linked alphabeta and alphaalpha by carboxymethyl cellulose (CMC) column chromatography in urea. Oxidized TM before fractionation, as well as the alphabeta and alphaalpha components, was found to have a molecular weight of about 80,000 daltons, indicating the disulfide bonds to be primarily intramolecular. Oxidation of dilute TM in 1 M NaCl by exposure to air also produced disulfide-linked alphabeta. Partially oxidized TM was found to separate into beta, alphabeta, alpha, and alphaalpha on CMC chromatography, and these were eluted with a linear gradient of NaCl at molarities of ca. 0.09, 0.11, 0.12, and 0.14 M, respectively. The oxidation process was investigated by CMC chromatography, and a possible mechanism is presented. The alphabeta and alphaalpha components may exist as dominant component in TM in vitro rather than as a random mixture of two subunits. A splitting of the electrophoretic band of the alpha subunit into a doublet was observed.


Structure and function of 5S ribosomal ribonucleic acid from Torulopsis utilis. III. Detection of single-stranded regions by digestion with nuclease S1.

Identification of single-stranded regions in Torulopsis utilis 5S RNA was attempted by the use of Nuclease S1,  a single-strand specific endonuclease. When T. utilis 5S RNA was subjected to prolonged incubation with Nuclease S1, about 50% of the substrate 5S RNA remained as large oligonucleotide "cores." Such Nuclease S1-resistant fragments were purified and sequenced by column chromatographic procedures. These analyses revealed that regions around positions 12, 40, 57, and 110 are in exposed single-stranded loops at 37 degrees C and that regions around positions 12 and 40 are most exposed at 20 degrees C. These results are compatible with our secondary structure model for T. utilis 5S RNA (Nishikawa & Takemura (1974) J. Biochem. 76, 935-947) except that the 5' part of the molecule (from the region around position 22 to that around position 57) might have a somewhat looser conformation than our secondary structure model suggests. The implications of such results are also discussed in relation to the presumed function of the sequence C-G-A-U-C (around position 40) as one of the recognition sites for initiator tRNA binding on ribosomes.


Reaction of DNA with glycidaldehyde. Isolation and identification of a deoxyguanosine reaction product.

Glycidaldehyde, a bifunctional carcinogenic alkylating agent, has been found to react with calf thymus DNA in vitro under alkaline conditions. A product was isolated and identified to be the result of a fusion of a delta4-imidazoline ring to the pyrimidine ring of deoxyguanosine at the N-1 and 2-NH2 positions. Following reaction and dialysis, the DNA was hydrolyzed enzymatically to nucleosides. The structural assignment of the product was based on comparison of the elution volume from a Sephadex G-10 column, the ultraviolet spectra at various conditions of pH, and the thin layer chromatographic properties with respect to an authentic sample. The formation of the extended ring system with DNA-derived guanosine may be related to the mode of carcinogenic action of this difunctional alkylating agent.


Isolation and properties of beta-D-galactoside-specific lectin from chick embryo thigh muscle.

A beta-galactoside-specific lectin, capable of agglutinating trypsinized rabbit erythrocytes, was isolated from 13-day-old embryonic chick thigh muscle and purified 1000-fold by affinity chromatography on asialofetuin/Sepharose and Sephadex G-100. A quantitative hemagglutinin assay based on the disappearance of single erythrocytes in a Coulter electronic particle counter was devised to measure lectin activity at different steps of purification. The molecular weight of the lectin was determined by gel filtration to be approximately 31,000, whereas polyacrylamide gel electrophoresis in sodium dodecyl sulfate gave a value of approximately 15,000, suggesting that the lectin is a dimer. The lectin is unstable below pH 5, and it requires the presence of dithiothreitol for the retention of maximal activity. The major portion of this lectin is membrane-bound; only 50% of the activity present in the muscle homogenate could be isolated in soluble form by extraction of muscle acetone powder with a buffer of high ionic strength. In view of the lack of a calcium requirement for its activity, the role of this lectin in myoblast fusion, a calcium-dependent phenomenon, is not clear.


Concerning the catalytic hydrogenation of polyprenyl phosphate-mannose synthesized by the retina.

Catalytic hydrogenolysis has been used as a means of distinguishing between classes of polyprenyl phosphate sugars which contain saturated alpha-isoprene units from those in which this grouping is allylic. Catalytic hydrogenation of dolichyl phosphate-[14C]mannose, synthesized by the retina of the embryonic chick was examined using different solvents and catalysts. When performed in chloroform:methanol (2:1), extensive cleavage to water-soluble radioactive material occurred due to hydrolysis caused by the reaction medium, even when buffered with tributylamine. When other solvents were used, considerable adsorption of radio-activity to the catalyst, in addition to variable amounts of hydrolysis, was encountered. Thus, the method is encumbered with artifact and extreme variability depending upon the reaction conditions employed, rendering it less than reliable for the purpose for which it was intended.


Ribonuclease inhibitor from human placenta. Purification and properties.

A soluble ribonuclease inhibitor from the human placenta has been purified 4000-fold by a combination of ion exchange and affinity chromatography. The inhibitor has been isolated in 45% yield (about 2 mg/placenta) as a protein that is homogeneous by sodium dodecyl sulfate-gel electrophoresis. In common with the inhibitors of pancreatic ribonuclease from other tissues that have been studied earlier, the placental inhibitor is an acidic protein of molecular weight near 50,000; it forms a 1:1 complex with bovine pancreatic RNase A and is a noncompetitive inhibitor of the pancreatic enzyme, with a Ki of 3 X 10(-10) M. The amino acid composition of the protein has been determined. The protein contains 30 half-cystine plus cysteine residues determined as cysteic acid after performic acid oxidation. At pH 8.6 the nondenatured protein alkylated with iodoacetic acid in the presence of free thiol has 8 free sulfhydryl groups. The inhibitor is irreversibly inactivated by sulfhydryl reagents and also by removal of free thiol from solutions of the protein. Inactivation by sulfhydryl reagents causes the dissociation of the RNase - inhibitor complex into active RNase and inactive inhibitor.


Rearrangements of integral membrane components during in vitro aging of sheep erythrocyte membranes.

In vitro aged sheep erythrocytes and sheep erythrocyte ghosts spontaneously release vesicles that consist of long protrusions affixed to flattened headlike structures. The intramembranous particles seen on the protoplasmic face of freeze fracture electron micrographs of vesicle protrusions are arranged in paired particle rows. On the equivalent fracture face of headlike structures, the particle density is low; if particles are present, they are clustered along the rim of the flattened headlike structure and at the junction with the protrusion. The released vesicles are depleted of the intramembranous particles seen on the exoplasmic face of ghost but retain almost exclusively particles of the protoplasmic face. Correspondingly, the exoplasmic face of ghosts that have released vesicles reveals a 28 percent higher density of intramembranous particles than that of fresh ghosts. Purified vesicles are depleted of spectrin but retain integral membrane proteins, with one of an apparent mol wt of 160,000 accounting for nearly 50 percent of the total protein (Lutz, H.U.,R. Barber, and R.F. McGuire. 1976. J. Biol. Chem. 251:3500-3510). When vesicles are modified with the cleavable cross-linking reagent [(35)S]dithiobis (succinimidyl propionate)at 0 degrees C, the 160,000 mol wt protein is rapidly converted to disulfide-linked dimers and higher oligomers. Exposure of intact ghosts to the reagent in the same way fails to yield equivalent polymers. A comparison of the morphological and biochemical aspects of ghosts and vesicles suggest that a marked rearrangement of membrane proteins accompanies the supramolecular redistribution of intramembranous particles during spontaneous vesiculation. The results also suggest that the paired particles of the protoplasmic face of vesicle protrusions are arranged in paired helices and contain the 160,000 mol wt protein as dimers.


New method for the isolation of Streptococcus mutans and its differentiation from other oral streptococci.

A new, improved agar medium for the isolation of Streptococcus mutans, the etiological agent of dental caries, was developed. In contrast to mitis-salivarius agar, this medium not only recovers a greater number of S. mutans strains from most oral specimens but, because of its mannitol and sorbitol content, it also facilitates the differentiation of S. mutans from other oral streptococci, e.g., S. salivarius, S. mitis, and S. sanguis, which do not grow or produce scanty growth only after 10 days of incubation. The medium is easy to prepare because of its simple and unique composition, is characterized by the presence of an acid indicator, and can be utilized under aerobic and anaerobic conditions as well. The medium cannot be used to distinguish among the eight serotypes, a to g and SL-1, of S. mutans. Mannitol-utilizing bacteria such as streptococci (e.g., S. faecalis) and other microorganisms (e.g., Staphylococcus aureus) are able to grow on this medium and can be distinguished from S. mutans by their unique colony morphology.


Isolation of Mycobacterium chelonei from a granulomatous lesion in a pig.

A rapidly growing, non-photochromogenic acid-fast organism was isolated from a lesion in a 6-month-old pig. The isolate was subsequently identified as Mycobacterium chelonei by its growth rate, but its failure to reduce nitrates, and by being arylsulfatase positive at 3 days.


A case study of embarrassment.

The psychoanalytic references to embarrassment are reviewed. Embarrassment, in the literature, is seen largely as an affect involving exhibitionistic and scopophilic conflicts and defenses against these. A case in which embarrassment was prominent is discussed. Embarrassment in the patient was an ego response which implied an external object for its manifestation. It involved exhibitionistic and scopophilic conflicts and projective defenses, but also operated in ego-gratifying and adaptive ways. Her embarrassment was understood through the analysis of an initial embarrassing dream of nakedness and other dreams and associated material as the defensive out-grouth of repeated exposures to the primal scene. Embarrassment was a resistance to remembering in the analysis, and the primal-scene experiences were partially reconstructed. The analytic situation was, in many ways, a symbolic re-creation of the primal scene, including the patient's response of embarrassment. The development of embarrassment in the patient's childhood was furthered and confirmed by its being an identification with the attitudes of both parents. Finally, some reflections on embarrassment and shame in its various forms are set forth.


Ultrastructural studies of mycobacterial phages isolated from the material of sarcoid patients.

Mycophages DNAIII, Legendre and Clark, isolated from sarcoid material by Mankiewicz, belongs with respect to their ultrastructural morphology, to the systematic group B according to Bradley or to group IV according to Tikhonenko. These phages contain 2-DNA, their head is a regular icosahedron, the tail consisting of a helix of protein subunits is attached to the head by a narrowed segment and is fixed in it by means of a disc-like structure. It is terminated by a ball-shoped or conical end structure. By their dimensions, these phages belong to the smallest ones in this systematic group. In comparison with other mycobacterial phages, the studied phages exhibit very low mechanical resistance. This characteristic is most pronounced in phage DNAIII. This group of mycophages is also very sensitive lipid solvents and increased temeprature. This phenomenon is also most pronounced in phage DNAIII. The explanation of these findings will be the subject of our further study.


Age-dependent changes in the oxygen consumption of the cerebral cortex, hypothalamus, hippocampus, and amygdaloid in rats.

The oxygen consumption of the cerebral cortex, hypothalamus, hippocampus, and amygdala, of both sexes, ranging in age from 21-805 days for male rats and from 21-780 days for the estrus and diestrus female rats, was measured. The oxidative activity of the hypothalamus, hippocampus, and amygdala decreased rapidly from Day 21 until the 4th mo. and stabilized till the 27th mo. in the hippocampus and amygdala and gradually decreased from the 12th mo. to the 27th mo. in the hypothalamus. The cerebral cortex showed a different pattern which kept a constant level from day 21 to 12 mo. of age. There was no statistically significant change in any of the regions at the time of vaginal opening. Low oxidative activity of the hypothalamus and amygdala in old age was still observed in castrated/hypophysectomized male rats compared with young ones. Therefore, the decreased oxygen consumption in old age seems to be due primarily to changes in the brain tissue itself and not due secondarily to changes in the gonadotropin and sex hormone levels. Female rats had higher oxygen consumption values than males in the cerebral cortex, the hypothalamus, and the hippocampus. In the amygdala the males had a higher consumption. Estrus female rats showed significantly higher oxygen consumption than diestrus females only in the hypothalamus.


A microhaemagllutination test in the diagnostics of syphilis.

The authors present the results of evaluation of a new, commercially produced haemagglutination test for the diagnosis of syphilis; they examined a total of 2 037 sera and compared the results with some classical and specific reactions for lues, used in the National Reference Laboratory for the Diagnosis of Syphilis in Prague. The authors clearly arrive at the conclusion that the haemagglutination test is a great contribution to the diagnostic lues which can be used in practice in the field with minimum expenditure and modest equipment.


Bacterial multiresistance to antibiotics. Study of types of bacterial resistance to antibiotics in the environment and in clinical material of a resuscitation ward.

Types of resistance were studied in clinical material and in the environment a resuscitation ward in the period from September 1973 to February 1974. Identical strains with the same types of resistance were isolated in both groups. They were verified by qualitative and quantitative tests of sensitivity to selected antibacterial substances. A high degeee of resistance to streptomycin, chloramphenicol, tetracycline and ampicillin was found in all gram-negative bacilli. High effectivity of colimycin, gentamicin and co-trimoxazole was confirmed in vitro. The strains of Staphylococcus aureus were fully sensitive only to oxacillin, lincomycin, spiramycin, vancomycin, gentamicin and co-trimoxazole. The strains of Streptococcus faecalis were found sensitive only to ampicillin and vancomycin.


Antigenic relations of Rickettsia prowazekii and Rickettsia canada, established in the study of sera of patients with Brill's disease.

Sera of patients with Brill's disease and of healthy persons with spotted fever in their past history were examined in the complement fixation reaction (CFR) to determine antigenic relations between R. prowazekii and R. canada. R. canada was found to have common antigenic determinants with R. prowazekii and R. mooseri. However, the antigenic determinants of R. canada differed from those of the mentioned rickettsiae. The titres of complement-fixing antibodies in the sera of patients with Brill's disease with the antigen of R. mooseri were lower than the titres with the homologous antigen within the range of 1-2 twofold dilutions of the serum. However, the oscillations of the titres with the antigen of R. canada in the study of the same sera were expressed in 1-5 twofold dilutions. In serological identification of canada rickettsiosis, antigens of rickettsiae of the spotted fever group should invariably be included in the investigation of the sera.


Levels of immunoglobulins and lysozyme in 10-year-old children from different regions and the condition of the lymphatic apparatus. I. Condition of tonsils in connection with some immunological parameters and different environment.

The condition of tonsils was investigated and the levels of serum IgG, IgA, IgM and lysozyme were measured in 305 ten-year-old children from three localities of the Central-Bohemian Region. The tonsils were not enlarged in 30.8% of children, in 26.2% they were enlarged, in 35.5% furrowed and in 7.5% of children tonsillectomy had been carried out. Mean levels of IgA were significantly higher in two localities in children with enlarged tonsils, insignificantly lower in children with furrowed tonsils and lowest in tonsilectomized children. The difference in the distribution of IgM levels were significant between girls with enlarged furrowed tonsils. The levels of lysozyme were not correlated with the finding in the tonsils, but a significant difference was found in the groups with enlarged tonsils between the three locailties under observation and in the groups with furrowed tonsils between two localities. The importance of the findings observed is discussed from the point of view of a possible influence of air pollution in two of the three localities under study.


Similarity of the carbohydrate structures of H-2 and Ia glycoproteins.

The glycopeptides produced by pronase digestion of two H-2K, two H-2D, and three Ia glycoprotein antigens were examined for size and charge. The glycopeptides derived from all of the antigens examined were found to have m.w. of 3250 +/- 200 daltons with a similar and variable composition of sialic acid residues. These data, when combined with the similarity in monosaccharide incorporation, suggest that the general parameters of the carbohydrate structure of the Ia glycoproteins from different I subregions and H-2 glycoproteins are highly similar if not identical.


Sera and the in vitro induction of immune responses. III. Adjuvant obtained from gliding bacteria with properties distinct from enteric bacterial lipopolysaccharide.

We have extended our studies on the role of bacterial contamination in the generation of fetal calf sera which support primary in vitro humoral responses by cultured mouse spleen cells. Gram-negative, gliding bacteria were isolated from a strongly supportive sample of fetal calf sera. Medium conditioned by the growth of these microorganisms had strong adjuvant effects in cultures of mouse spleen cells supplemented with a non supportive, deficient sera. The adjuvant and mitogenic activities of the bacterial conditioned medium were then compared to those of bacterial lipopolysaccharide from Salmonella typhosa 0901 in cultures of LPS responder and LPS nonresponder spleen cells. From the results of these comparative studies, we conclude that the active factor(s) obtained from the gliding bacteria is an adjuvant with properties very distinct from those of highly purified enteric bacterial LPS.


Serodiagnosis of hepatitis B virus infection by antibody to core antigen.

In a military population antibody to hepatitis B core antigen (anti-HBc) was found in 39% of acute hepatitis cases negative for hepatitis B surface antigen (HBS Ag) and in 96% of HBs Ag-positive cases. Persistence of antibody to HBs Ag (anti-HBs) in convalescent-phase sera was significantly greater (P less than 0.001) in individuals with acute HBs Ag-positive hepatitis B than in patients with clinical HBs Ag-negative hepatitis B. The prevalence of anti-HBc in the absence of HBs Ag, anti-HBs, and clinical disease was 3.2% in this military population. In longitudinal studies of hepatitis B  infection, the presence of anti-HBc preceded anti-HBs and improved the ability to determine the onset of sublicnical infection. Anti-HBc is a useful serologic marker for the study of the epidemiology of hepatitis B and improves the efficiency of detection of hepatitis B virus infection.


Mouse teratomas and embryoid bodies: their induction and differentiation.

Teratomas were induced by the transfer of mouse blastocysts (C3H and 129/J strains) and egg-cylinders (C3H) to extra-uterine sites. C3H and 129/J blastocysts cultured in vitro for 4 or 5 days could also form teratomas in extra-uterine sites. Four transplantable teratomas, or teratocarcinomas, were derived from C3H embryos; embryoid bodies were derived from each line. The differentiative capacity of a teratocarcinoma was shown to be similar whether it was maintained as a solid tumour or as embryoid bodies.


Differentiation of primordial germ cells in the embryonic development of Thermobia domestica, Pack. (Thysanura): an ultrastructural study.

The primordial germ cells(PGCs) of Thermobia domestica undergo some morphological changes during the embryonic development. Most conspicuous are the changes in the ultrastructure of the nucleus, whose envelope shows a high degree of activity. Two types of vesicles bled off from the nucleus; the ones with the light interior are called the accessory nuclei, the others, with electron-opaque contents, have been termed the dense bodies. The nucleolus, initially clustered at the nucleus centre, undergoes dispersion and assembles again towards the end of embryonic development. At the same time, the sex differentiation of PGCs takes place. It is preceded by an increase in the activity of Golgi complexes and in the volume of lysosomes and lamellar bodies, the latter giving rise to lipid droplets. At the early stages of postembryonic development, preoogonia and prespermatogonia can readily be distinguished. Preoogonia have a wavy-surfaced nucleus and their cytoplasm contains dense bodies. In prespermatogonia, the nucleus is spherical with smooth envelope and there are no dense bodies in the cytoplasm. Throughout the period studied there occur nucleolus-like bodies and nuage material considered to be the germ-cell determinants in this species.


Properties of extra-embryonic ectoderm isolated from postimplantation mouse embryos.

Extra-embryonic ectoderm isolated from the mouse embryo as late as 81/2 days post coitum can form cells with the morphological characteristics of trophoblast giant cells both in ectopic sites and in vitro. This similarity to the properties of ectoplacental cone tissue provides further support for the postulated common origin of both tissues from the trophectoderm of the blastocyst.


Effects of prenatal administration of hadacidin, a cancer chemotherapeutic agent, on the development of hamster fetuses.

A single intraperitoneal injection of 1-2-5 g/kg body weight of hadacidin was teratogenic when administered to pregnant hamsters between days 8 and 11 of gestation. Both the frequency of malformation and resorption were related to the dose and time of hadacidin administration. The drug produced both gross and microscopic malformation in the fetus and impaired its general growth. The malformation involved craniofacial structures, central nervous system, respiratory, digestive and urinary systems, limbs and tail. Morphologically different types of cleft lip and cleft palate were related to the dose and time of hadacidin treatment. An association was observed between cleft lip, cleft palate and micrognathia on one hand, and between cleft palate and micrognathia on the other. Significance of association between the lip, the palate and the mandibular malformation was discussed and the hypothesis that human cases of Pierre Robin syndrome may result from an environmental assault was supported. It was suggested that the fetal weight and the microscopic analysis should be included in the criteria for the teratological drug safety evaluation procedures.


Replication of minute virus of mice in Chinese hamster ovary fibroblasts.

Minute virus of mice (MVM), a parvovirus, replicates in Chinese hamster ovary (CHO) cells. Although replication of the virus cannot readily be detected by haemagglutination, it can be measured by plaque assay on mouse strain LM cells.


Heterogeneity among strains of Mycoplasmatales virus-laidlawii 2.

Five isolates of Mycoplasmatales virus-laidlawii 2 (MV-L2) derived from bovine strains of Acholeplasma laidlawii were shown to differ in host range, plaque morphology and neutralization tests with MV-L2 antiserum. Cross-testing using virus resistant clones of A. laidlawii confirmed the heterogeneity of this group. Adaptation of viruses to sub-optimal host was demonstrated.


Interaction of non-specific inhibitor and rabies virus haemagglutinim.

In order to elucidate the properties of an inhibitor of rabies virus haemagglutinin in normal animal sera, experiments were made with the HEP Flury strain and calf serum which contains the inhibitor. The results of physico-chemical treatment, gel-filtration and density analysis suggested lipoprotein involvement. When inhibitor and haemagglutinin were mixed, the separate activities could be recovered from the mixture by centrifuging on a sucrose density gradient. By contrast, neither haemagglutinin nor inhibitor could be recovered by this treamtnet when the inhibitor was added at the start of virus growth. The binding of inhibitor with rabies virus during virus growth seems irreversible and different from the binding of inhibitor with pre-formed rabies haemagglutinin.


Malonaldehyde content of food.

Several types of commercially available food both cooked and uncooked were tested for the presence of the carcinogenic initiator and mutagen, malonaldehyde, which is a breakdown product of unsaturated fatty acids. The thiobarbituric acid derivative of malonaldehyde from meat extract was identified either by thin layer chromatography or by gas chromatography after silylation. Malonaldehyde was also identified directly by gas chromatography. Among the meats purchased from supermarkets, beef had the greatest amounts of malonaldehyde. Turkey and cooked chicken had high levels. Most cheeses had only small amounts of malonaldehyde. In contrast, many vegetables and fruits had either minute amounts or no malonaldehyde.