A comparative study of the Crimean hemorrhagic fever-Congo group of viruses. Twelve strains of the Crimean hemorrhagic fever (CHF)-Congo group of viruses the Bunyaviridae family were investigated with respect to sensitivity to lipid solvents and temperature, pathogenicity for animals, interactions with cell cultures and antigenic relationships. Complement fixation, agar gel diffusion and precipitation, immunofluorescence and neutralization tests showed Hazara virus to have a number of features distinguishing it from the other antigenic type of the CHF-Congo group.


Resolution of human fibroblast interferon into two distinct classes by thiol exchange chromatography. Chromatography of crude and purified human fibroblast interferon on activated thiol Sepharose 4B yielded two classes of material exhibiting different stability to inactivation by mechanical stress. Interferon which bound to the gel accounted for 20--30 per cent of recoverable activity, could be eluted with reducing agents and was unstable. Unbound material did not bind on subsequent rechromatography and was stable to shear forces. Leucocyte interferon did not bind, nor did fibroblast interferon treated with thiol blocking reagents. Peak eluted fractions had a specific activity of up to 10(7.9) units per milligram of protein, representing a 1700 fold purification.


Factors involved in the age-dependent resistance of mice infected with low-virulence mouse hepatitis virus. Four-week-old weanling mice survived, whereas 1-week-old suckling mice died, after intraperitoneal inoculation of mouse hepatitis virus, MHV-S strain. The factors involved in this difference in susceptibility were studied. After virus inoculation, differences in virus growth in the liver and spleen were observed, which correlated with the susceptibility of animals to the virus. Interferon, detected at an early stage of infection, was considerably lower in suckling mice than in weanling mice. Titers of MHV-S in peritoneal cells from infected animals were at least 100 times greater in suckling than in weanling mice, and a similar, but less prominent difference in virus growth was also found in the corresponding cultured macrophages. After transfer of peritoneal cells from weanling to suckling mice, a decrease in mortality of infected suckling mice was observed. These results suggest that both interferon and macrophages may be important in the age-dependent resistance of mice to MHV-S infection.


Properties of Border disease virus as studied in a sheep cell line. A fetal lamb muscle cell line has been isolated in which two strains of Border disease virus replicate well and induce a clear-cut cytopathic effect, thus providing a sensitive and practical assay system for both the virus and neutralizing antibodies. This system enabled us to study some characteristics of Border disease virus in comparison with two other agents responsible for hog cholera and bovine viral diarrhea. Our results indicate that the last virus and the Border disease agent are indistinguishable in vitro.


Immune retention: immunological requirements for maintaining an easily degradable antigen in vivo. Radioactive human serum albumin (125I-HSA) was injected into the hind foot pads of unimmunized mice, actively immunized mice and mice passively immunized with mouse or rabbit anti-HSA serum. Eleven days later the unimmunized mice had cleared most of the 125I-HSA. In contrast, a high concentration of 125I-HSA was retained in the feet and draining lymph nodes and, to a lesser extent, in the spleen of the actively or passively immunized mice. Although immune retention required specific antibody, it appeared to be independent of T-cells or T-cell factors, since passively immunized nude mice retained antigen as well as actively or passively immunized normal mice. Depletion of the complement system with cobra venom factor (CVF) increased antigen retention in the feet but decreased retention in the spleen. Treatment with CVF did not decrease antigen retention in lymph nodes of actively immunized mice. Such treatment did, however, decrease retention in lymph nodes of passively immunized mice although not to the same extent as in the spleen. Retention of antigen in the feet was not only complement-independent but was also Fc independent, since F(ab')2 fragments of IgG could mediate immune retention. Antigen dose response studies indicated that immune retention in lymph nodes occurred optimally with minute amounts of antigen, whereas optimal retention in the feet required much higher concentrations of antigen. Foot pad injections of non-radiolabelled HSA eliminated 60% of the radioactivity retained in the foot pads of immunized mice. In contrast, non-radioactive egg albumin (EA) had almost no effect on retained HSA. However, if the mice were immunized to both EA and HSA, an injection of EA would displace a significant amount of retained HSA. Complexes of one specificity can apparently displace some retained antigen of a differing specificity.


Biophysical and serological characteristics of some New Zealand bovine enteroviruses. Five previously reported New Zealand bovine enteroviruses are further characterised with regard to their biophysical and serological properties. Characteristics studied included heat stability, buoyant density, virus replication, plaque production and thin section electron microscopy. Serological tests indicated the isolates belonged to two different serogroups which showed only limited relationships with U.S. serotypes BES 1, 3 and 4. A correlation was noted between 2(alpha-hydroxybenzyl) benzimidazole sensitivity, heat stability and the serogroup of the isolates.


The pharmacodynamics of 4-(p-chlorophenylthio)butanol (W-2719) in humans. 4-(p-Chlorophenylthio)butanol (W-2719) is absorbed, metabolized and excreted when administered orally to human volunteers. The butyrate (W-2718) and the acetate (W-2683) metabolites were found in plasma with W-2683 being the major biotransformation product and detectable seven days after administration with an apparent half-life of 33.2 h. p-Chlorothiophenol appeared to be the major biotransformation product in urine and was detectable through day 6. As in plasma, the other major biotransformation product was W-2683, which was again detectable through day 7 and had a t 1/2 in urine of 40.8 h. The minor metabolite found in urine was W-2718. No W-2719 was found in either plasma or urine after oral administration.


Early intrathecal synthesis of interferon in herpes encephalitis. Large quantities of type I interferon were detected in the cerebrospinal fluid (CSF) collected at the onset of herpes encephalitis. This interferon was synthesized intrathecally and disappeared about 10 days after the beginning of neurological signs. In 12 cases of post-eruptive measles encephalitis and in four post-rubella encephalitis, type I interferon was present only in low amounts, or not found at all, even in the CSF collected early. The existence of an intrathecal synthesis of interferon during encephalitis provides a valuable contribution to both the early diagnosis of herpes encephalitis and the study of the pathogenesis of virus infection of the central nervous system (CNS).


Pathophysiology and pharmacotherapy of spasmodic torticollis: a review. The few existing neuropathological, neurochemical, and neuropharmacological studies have shed little light on the pathophysiology of spasmodic torticollis (ST). The relevance of experimental ST in animals and drug-induced ST in man to idiopathic ST is unclear. Most pharmacotherapeutic endeavors have focused on drugs affecting basal ganglia function. Unfortunately, problems of sample size, clinical heterogeneity of patient population, research design, objective evaluation of response, documentation of key data, and adequacy of duration of follow-up make interpretation of published results difficult. Because of the heterogeneity of ST, investigations aimed at establishing a neurotransmitter profile for each patient by observing the acute response to a test dose of drugs affecting cholinergic, dopaminergic, serotonergic, and gamma-aminobutyric acid systems may provide a more rational basis to the selection of treatment.


Correlation of a specific chromosomal marker, 21q-, and retroviral indicators in patients with thrombocythemia. A chromosomal anomaly, 21q-, has been found in association with retroviral indicators in patients with myeloproliferative disorders (MPD) including polycythemia vera (PV), essential thrombocythemia (ET), chronic myelocytic leukemia (CML) and acute non-lymphocytic leukemia (ANLL). The viral indicators are found in platelet homogenates of thrombocythemic patients. Evidence is presented from 2 laboratories (Philadelphia, USA and Bologna, Italy) for the 21q- deletion in MPD patients. Thirty patients evaluated for the presence of both viral and chromosomal markers in Philadelphia showed positive correlations.


Synthesis of Plasmodium knowlesi polypeptides in a cell-free system. Preliminary results are presented concerning the characterization of Plasmodium knowlesi antigens. RNA with messenger activity has been extracted from P. knowlesi and translated in a rabbit reticulocyte cell-free system. The products of this system have been analysed both before and after precipitation with specific P. knowlesi antiserum and many polypeptides ranging in molecular weight from over 100 000 to approximately 20 000 have been found. A good correlation was found between these results and those obtained by McColin with in vitro labelling of P. knowlesi with ((3)H)-isoleucine. The major polypeptide in both systems had a molecular weight of about 50 000-55 000. These results indicate the parasite origin of the antigens.


On the nature of Romanowsky dyes and the Romanowsky-Giemsa effect. This paper reviews the nature of Romanowsky staining and the relationship between Romanowsky dyes and the Romanowsky-Giemsa effect (RGE). On blood and bone marrow smears the RGE is characterized by a purple colouration of nuclei and neutrophil granules. The nuclear purple contrasts strongly with the blue cytoplasmic staining of cells rich in RNA. Requirement for the occurrence of RGE are: I A cationic dye: The best dye is azure B and, though azure A gives the nuclear purple colour, the cytoplasmic blue is inferior. No other cationic dye such as methylene blue is suitable. 2 An anionic dye: Most commonly eosin Y is used, but it can be replaced by the erythrosins. Full halogenation of the fluorescein (four atoms of bromine or iodine) is not necessary. Phloxine and rose bengal are unsuitable. 3 An appropriate substrate: These are proteins with acidic side groups or proteins bound to a polyanion. For the interaction with the dyes substrates must provide a suitable three-dimensional network which is why the RGE is not obtained in solutions. A tentative theory of RGE is advanced and briefly discussed.


Comparison of several in vitro assay methods for the quantitative determination of complement consumption ('binding') by gammaglobulin preparations. Unspecifically induced activation of Complement (C) in solution can be caused either by denatured Ig molecules and by polymeric aggregates of same or more specifically by antigen-antibody complexes. As a quality criteria, e.g. for i.v.-IgG preparations, it is agreed by the majority of people concerned that the activation of C via the classical pathway might not be unspecifically initiated. Therefore, the generally used assay systems are based on quantitating the degree of consumption of C undergoing an antigen-independent, 'frustrated' activation via the classical pathway. There are principally two different types of test modifications which were investigated: a) C is being kept constant and protein is diluted or b) protein is being kept constant and C is titrated. When a group of differing IgG preparations or IgG fragments was assayed for its so-called 'anticomplementary' activity in various already described test methods, the particular test results from the individual test methods could not unequivocally be compared with each other because some of the methods had not been optimized with regard to the amount of reagents or test substance used; only a rough estimate of the data was obtained. The value of the not standardized assays remains, therefore, questionable reasonably good and reproducible results are obtained when the amount in particular of C added as well as its quality have been adequately optimized and standardized. This allows a refined differentiation of various IgG preparations by a reliable and unadjusted quantitation of C consumption. If for example, inappropriate and not optimized rations of C amboceptor (e.g. slight C excess) was used, a suppressed C consumption was found. It seems advisable to accept prophylactic testing of anticomplementary activity for quality control as one out of several other in vitro and in vivo parameters which potentially might predict safety for the patient, the physician, and also for the producer. However, it is agreed that, irrespective of in vitro tests, in vivo testing in animals and clinical proof of safety is undoubtedly needed.


An unstable allele of the am locus of Neurospora crassa. The mutant strain am126 was isolated, using the direct selection procedure, after nitrous acid mutagenesis. It produced neither measurable NADP-dependent glutamate dehydrogenase (GDH) nor immunologically cross-reacting material. That the am126 strain produced some form of GDH product was shown by the fact that it complemented several other am mutant strains. The GDH formed by complementation between am126 and each of two other am mutants was relatively thermolabile, but could not be distinguished from wild-type GDH formed by electrophoresis in polyacrylamide gels. This, together with the relatively high yield of the complementation enzymes, suggest that the am126 product is a polypeptide chain not grossly abnormal in structure. The spontaneous revertant frequency was between 0.3 and 3 prototrophic revertants per 10(5) live cells. This frequency was at least 40 times greater than that for am19, which had the second highest spontaneous revertant frequency among the mutants tested. Neither meiosis nor mutagenesis increased the revertant frequency, nor did incubation at elevated temperatures lower it. Sixty-eight revertant strains were examined for thermostability of their GHD. All appeared to be identical to wild type. Seven of the revertant strains were also tested for instability with regard to forward mutation to am auxtrophy. None was found to be unstable. Models for the genetic instability of the am126 mutation are discussed.


Genetic modulation of RNA metabolism in Drosophila. IV. Measurement of rats of RNA synthesis by density labeling. Accumulation of RNA was measured in adult males of two genotypes: car bb/Ybb- and car bb/YbbSuVar-5. The two genotypes have similar amounts of rDNA, which is reduced in comparison to wild type (CLARK, STRAUSBAUGH and KIEFER 1977). Although genotypically bobbed, car bb/YbbSuVar-5 flies have a wild-type phenotype; car bb/Ybb- flies are both phenotypically and genotypically bobbed (CLARK, STRAUSBAUGH and KIEFER 1977). The wild-type phenotype observed in the car bb/YbbSuVar-5 flies is thought to be the result of an increased rate of rRNA synthesis due to the presence of the YbbSuVar-5 chromosome (SHERMOEN and KIFFER 1975; CLARK, STRAUSBAUGH and KIEFER 1977; CLARK and KIEFER 1977). To further define this phenomenon, the absolute accumulation of RNA was measured in the two genotypes, using density labeling methods. The accumulation of RNA is 1.4 to 1.8 times higher in car bb/YbbSuVar-5 flies than in car bb/Ybb- flies, demonstrating that there is genetic regulation of synthesis in this genotype. The use of density-labeled nucleosides has clearly shown that there is no difference in precusor pool sizes or use between the two genotypes studied.


Release of particle associated RNA dependent DNA polymerases by primary chicken and quail embryo cells. These results show that primary chick embryo cells in culture release a particle bound RNA dependent DNA polymerase into the culture fluid. The enzyme is also found in the allantoic fluid of embryonated chicken eggs. This phenomenon is not restricted to the chicken system because primary quail embryo cells which are not known to express the so far known endogenous viruses release a particle bound RNA dependent DNA polymerase as well. In both cases this enzymes may be part of an hitherto unknown endogenous viral system. However, since the expression of the chicken- and quail-enzyme goes through a sharp peak during embryogenesis one may speculate on a physiological role for the newly described enzymes during development.


Cellular and virological studies directed to the pathogenesis of the human myelogenous leukemias. 1. Work over the past years and especially results of the past few years indicate that type-C viral or viral related genetic information exists in humans. 2. We do not know how this information entered humans or whether it causes disease, but it is of interest that the probes from the viruses used to detect this information are from the very same viruses which we find can affect growth and differentiation of some human hematopoietic cells. 3. The status of actual virus isolates from humans, though encouraging because of similarities of isolates from five different laboratories, remain very perplexing and so far have not been especially informative to human leukemogenesis. 4. In the near future we hope to clone in bacteria the viral related sequences detected in human DNA in order to more precisely determine their chemical and biological properties. The HL-60 system may also afford an opportunity to purify receptors for CSF. When CSF and other, perhaps more important, regulatory factors are purified, we would like to determine if they bind differently to leukemic and normal cells.


Placental alkaline phosphatase, alphafetoprotein and carcinoembryonic antigen in testicular tumors. Tissue typing by means of cytologic smears. Indirect immunofluorescence and radioimmunoassay with specific rabbit antisera demonstrated the occurrence of alphafetoprotein (AFP), carcinoembryonic antigen (CEA) and placental alkaline phosphatase (PLAP) in primary testicular tumor cells. Embryonal carcinomas had AFP- and CEA-containing cells, sometimes PLAP. PLAP and sometimes CEA were found in seminoma cells. Sera from patients with advanced non-seminomatous tumors could contain any of these antigens or any combination of them. Sera from patients with seminomas had raised PLAP or CEA. PLAP appears to be a new marker for seminoma.


Mouse mammary tumor virus (MMTV) antigen(s) are present on B lymphocytes of BALB/c mice. Immunofluorescence studies using a polyvalent anti-MMTV serum revealed the presence of MMTV antigen(s) on the surface of spleen cells from the low mammary tumor incidence mouse strain BALB/cCrgl. Upon separation of the splenocytes on nylon-wool columns the populations of cells exhibiting positive fluorescence were nylon-adherent. These cells were considered to be B lymphocytes by the following criteria: presence of surface immunoglobulin (Slg), absence of Thy-1 antigens, enhanced responses to B-cell mitogens and decreased reactivity to T-cell mitogens. In order to ascertain the specificity of the cell surface immunofluorescence reaction, blocking studies were performed. Immunofluorescence was blocked by preincubation of the antiserum or with purified MMTV, but not by R-MuLV. Non-specific binding of the anti-MMTV to Fc receptors present on B lymphocytes was ruled out by the use of Fab'2 fragments of IgG from the various sera. Double label immunofluorescent studies with rhodamine-conjugated anti-mouse Slg Fab'2 fragments and rabbit anti-MMTV Fab'2 with fluorescein-conjugated goat anti-rabbit Fab'2 showed the simultaneous appearance of both labels in the nylon-adherent splenocytes. These results suggest the expression of viral polypeptide(s) encoded by an endogenous MMTV on the surface of B-type splenocytes in mice devoid of intact viral particles.


Expression of feline xenotropic RNA tumor virus in hybrids between permissive human and non-permissive mouse cells. Somatic cell hybrids were generated by fusing human (A549) cells, cloned after infection with the feline xenotropic CCC virus, to mouse (3T3) cells which are non-permissive for this virus. Hybrid clones were found to be capable of expressing infectious virus. CCC virus expression, however, was regulated in the hybrid cells in such a way that 20-200 times less virus was released into the culture fluid than by the human parental line. Thus human permissiveness for this virus is co-expressed with murine restriction. Markers for twenty human chromosomes were assayed in the hybrid clones. No single human chromosome was found to be essential and sufficient for CCC virus production, since none of them was consistently present or lost in virus-positive and virus-negative clones, respectively.


Oral rehabilitation by prosthetic restoration after maxillectomy for malignant tumors. In maxillectomy patients, speech intelligibility between before and after insertion of prostheses has been reviewed by many investigators. Speech intelligibility was not related to the size of the defect, but rather to the defect region. Speech intelligibility was considerably improved by the use of prostheses. A great difference could not be found in escape rate from the nasal cavity between before and after insertion of prostheses, but there was a remarkable change in the spirometer. Postmaxillectomy functions are also easily restored by prosthetic oral rehabilitation, necessary to aid surgical procedure.


Simian rotavirus SA-11 plaque formation in the presence of trypsin. Incorporation of 5 micrograms of trpsin per ml of the overlay (Eagle minimal essential medium-0.7% Ionagar no. 2) was found to be necessary for plaque formation by simian rotavirus SA-11. Plaques of 3 to 4 mm in diameter were produced in MA-104 cells after 5 days of incubation at 37 degrees C. Plaque size was even larger (5 to 6 mm) in monolayers of African green monkey kidney cells. Addition of diethyl-aminotheyl-dextran, protamine sulfate, or 5-bromodeoxyuridine to the trypsin-containing overlay did not improve plaque formation by the virus. Incorporation of beef extract or yeast extract to a final concentration of 0.5% in the trypsin-containing overlay inhibited plaque formation. On the other hand, the presence of lactalbumin hydrolysate or peptone at a similar concentration in the overlay did not inhibit plaque formation. When methylcellulose was used instead of the agar as the solidifying agent in the overlay, no plaques were seen. SA-11 is a useful model for the study of human rotaviruses, and this relatively simple plaque assay system should further enhance its usefulness in this regard.


Lectins in diagnostic microbiology: use of wheat germ agglutinin for laboratory identification of Neisseria gonorrhoeae. A lectin slide agglutination test has been developed for the confirmatory identification of Neisseria gonorrhoeae. With wheat germ lectin as an agglutinin, 164 of 165 clinical isolates of N. gonorrhoeae gave a 3 to 4+ reaction within 6 to 8 min. Four gonococcal isolates, even though negative by the fluoresecent-antibody method, gave strong positive reactions with the wheat germ lectin. Among 23 isolates of Neisseria meningitidis tested, which included representatives of sero-groups A, B, C,D, X, Y, and Z, only one strain in group X gave a false-positive reaction. The nonpathogenic species of Neisseria, as well as Branhamella catarrhalis, all showed negative reactions with the wheat germ agglutinin. The novel method provides a simple, rapid, and inexpensive means for the laboratory diagnosis of gonorrhea and obviates the need for performing second-stage sugar fermentation studies or utilizing the more expensive fluorescent-antibody techniques.


Virus-like particle, 35 to 40 nm, associated with an institutional outbreak of acute gastroenteritis in adults. In an outbreak of acute gastroenteritis in an institution for the mentally retarded at Otofuke, Hokkaido, Japan, virus-like particles were observed by electron microscopy in five of seven stool specimens from the patients. The particles had 10 rod-shaped and 10 round projections or capsomeres on the periphery, measured 35 to 40 nm in diameter, and had a buoyant density of 1.35 to 1.37 g/ml in cesium chloride. Attempts to culture these particles in tissue culture or in mouse brain were unsuccessful. Immune electron miscroscopy performed with the virus from the patients as antigen demonstrated significant serological responses in all patients examined. Antigenic similarity of the virus particles obtained from five patients was also confirmed by immune electron microscopy with the paired acute- and convalescent-phase sera of one of the patients. Furthermore, in immune electron micrsocopy these particles appeared to have no antigenic relationship to three candidate viruses for gastroenteritis so far reported: the Norwalk agent, the W agent, and the calicivirus-like particle. These results suggested the possibility that this agent, tentatively designated as the Otofuke agent, might be a new candidate virus for gastroenteritis.


Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain. The sensitive peroxidase-antiperoxidase (PXAPX) method individually and in conjunction with the Papanicolaou (PAP) stain was used to detect herpes simplex virus (HSV) in specimens from human female genitalia. Initial studies using a model system of HSV-1 or HSV-2-infected Vero cells established (i) acetone as the most effective fixative, (ii) optimal dilutions of preimmunization and anti-HSV serum for differentiation of infected from noninfected cells, (iii) optimal concentration of 3,3'-diaminobenzidine tetrahydrochloride and H2O2 for maximal staining of infected cells with minimal background reaction, and (iv) removal of endogenous peroxidase with absolute MeOH. These various parameters, once established, were utilized in the PXAPX or PXAPX-PAP on human specimens from the vulva or cervix. In these specimens, examined by standard light microscopy, PXAPX-positive cells were dark brown with a single nucleus or multiple nuclei. By coupling the PAP to the PXAPX, detailed nuclear observations of PXAPX-positive cells were possible and revealed nuclear changes consistent with HSV infection, including syncytium formation, chromatin condensation, and an occasional Cowdry type A inclusion. The PXAPX and PXAPX-PAP correlated (r = 0.742) over a period of 72 h with HSV isolation from these samples.


Cytochemical analysis at the fine-structural level of trypanosomatids stained with phosphotungstic acid. The ethanolic phosphotungstic acid (PTA) technic was used to detect, at the fine-structural level, basic proteins in various developmental stages of pathogenic Trypanosoma cruzi, and nonpathogenic Herpetomonas samuelpessoai, Leptomonas samueli, and Crithidia deanei, trypanosomatids. Reactions were observed in the nucleus of all stages. In the kinetoplast of epimastigote and promastigote forms reactions were noted mainly at the periphery. In trypomastigotes and choanomastigotes forms, however, an intense reacion was observed thorughout the kinetoplast. Reactions were present in cytoplasmic vesicles related to protein storage in T. cruzi and in membrane-bounded peroxisome-like organelles of H. samuelpessoai, L. samueli and C. deanei. The network of filaments which forms the paraxial rod did not react. In the flagellum, reaction was noted only at the peripheral doublet microtubules. PTA reacts also with structures related to the junction between the flagellar and cell body membranes.


[Periodic, metabolic and structural phenomena in a protist, Euglena gracilis]. Sychronous divisions of Euglena gracilis strain Z can be obtained by various methods. When the cells are cultivated in a medium containing lactate as the sole carbon source, synchronous divisions are observed, independent of the conditions of illumination. Nevertheless, there exists a relationship between the phase of cell division and ther periods of light and darkness applied to the culture. During the cell cycle, the synthesis of macromolecules is discontinuous--this is true of nuclear and mitochondrial DNA, ribosomal and nonribosomal RNA, and certain proteins (cytochrome c 558). Cyclic variations in the structure of mitochondria and chloroplasts are observed. In the course of the cell cycle, sequential metabolic processes accompany structural modifications of the organelles. Also, at the beginning of the cycle, at the start of phase G1, the cytoplasmic ribosomes are synthesized, and then, in green euglenids, nonribosomal RNAs are formed. These syntheses of RNA precede enlargement of the chondriome and plastids. In mid-G1 phase, a new synthesis of RNA begins, which precedes synthesis of nuclear and mitochondrial DNA. At the end of G1 phase, division of organelles starts, beginning with the chondriome and plastids, arranged in a network.


Effects of hydroxyurea on Crithidia fasciculata. Hydroxyurea (HU) inhibits increase in cell number in cultures of Crithidia fasciculata. Complete inhibition is produced by 8 mM and higher concentrations. If HU is not removed, population growth resumes in 45-50 h; if HU is removed, partially synchronous growth occurs through 2 cycles. During HU inhibition, the rate of DNA synthesis is reduced to 1% of that in exponentially growing cultures; protein and RNA syntheses continue at slightly reduced rates. Mean cell size and protein and RNA contents per cell increase; rate of oxygen consumption per mg cell protein remains constant. The behavior of a culture upon addition of HU and upon its removal agrees with predictions based on the hypothesis that the only direct effect of HU is to block DNA synthesis. The synchrony produced by HU is judged satisfactory for investigations of kinetoplast and nuclear replication but not for biochemical characterization of other aspects of the cell cycle.


Poly(A)+ RNA from Tetrahymena: stimulation of protein synthesis in vitro. Cell-free synthesis of high molecular weight polypeptides, programmed by RNA from Tetrahymena pyriformis strain W is reported, and methods for preparation of the RNA are described. The RNA was extracted by the SDS-phenol-chloroform-isoamyl alcohol technic. The bulk of extracted RNA was ribosomal and on sucrose gradients peaked at approximately 17S and 25S. After heat denaturation all the 25S RNA was converted to 17S, indicating the presence of hidden breaks, possibly the result of nuclease activity during extraction. Nevertheless, when poly(A) +/- RNA was collected using oligo-(dT)-cellulose column chromatography, it promoted a 15-fold increase in incorporation of [35S] methionine into TCA-precipitable material. Slab-gel electrophoresis and autoradiography of the product revealed 12 different major polypeptides, varying in weight from 28,000 to 65,000 Daltons. A method for preparation of translatable RNA from Tetrahymena will make possible the comparison of messenger RNAs associated with specific cell structures and with different developmental events.


Interferon induction with Newcastle disease virus in FS-4 cells: effect of priming with interferon and of virus inactivating treatments. Inoculation of human FS-4 cells with Newcastle disease virus (NDV) resulted in the induction of two distinct interferon responses, one that peaked at about 5 hr (early response) and one that reached a maximum between 10 to 24 hr after inoculation (second response). The early interferon response was enhanced by previous treatment of the cells with interferon (priming), whereas the second response decreased after interferon treatment in a dose-dependent manner. The early response diminished with decreasing multiplicities of infection, the magnitude of the second response in unprimed cells was relatively independent of the dose of NDV employed. The early interferon response was sensitive to inhibition by actinomycin D for only 1 hr after inoculation. In marked contrast, the second response remained sensitive to inhibition by actinomycin D until 12 hr after inoculation. The ability of NDV to induce the second response was greatly diminished by irradiation of the virus with ultraviolet light or by its treatment with hydroxylamine, whereas the ability to stimulate the early response was relatively resistant to these virus-inactivating treatments. Treatment of NDV with hydroxylamine abolished the virus to induce the second response at the same rate as it destroyed infectivity. The results suggest the existence of at least two distinct mechanisms of interferon induction by NDV; the early response is triggered either by a virion component or by a product of primary transcription, whereas induction of the second response requires the expression of some functions of the virus not needed for triggering the early response.


Induction of SV40-related transplantation immunity in mice by BK virus-transformed cells. The reactivity of tumor-specific transplantation antigens (TSTAs) induced by SV40 and BK virus (BKV) was studied in Balb/Ccr mice with SV40-transformed tumor cells (mKSA-4AC). The mice, which had received BKV-transformed mouse or hamster cells, were highly resistant to the challenge with mKSA-4AC. The mKSA-rejected animals, however, were as susceptible as non-immunized ones to the subsequent challenge with Moloney sarcoma virus (Kirsten)-transformed cells. The result shows that BKV-transformed cells have a TSTA immunologically related closely to that induced by SV40.


Fatal interstitial pneumonitis induced by low dosage of bleomycin in an esophageal cancer patient. One-shot intratumor injection of 60 mg oil-bleomycin prior to subtotal esophagectomy induced fatal, acute interstitial pneumonitis in a 53 year-old esophageal circinoma patient. The progression of bleomycin-induced pneumonitis was rapid and the patient succumbed to respiratory insufficiency 41 days after surgery (58 days after bleomycin administration). Autopsy revealed bilaterally increased lung weight and the organ was of rubbery consistency and anthracotic. Histology showed advanced interstitial fibrosis, numerous lymphocytes in alveolar septi and fibrinous exudate, macrophages and desquamative cells within the alveolar spaces. In the present case, pneumonitis was due not to patient age or dosage level of bleomycin, but rather to the administration method.


The errors in assembly of MuLV in interferon treated cells. Interferon treatment of JLSV-6 cells chronically infected with Rauscher MuLV leads to the formation of noninfectious particles (interferon virions) containing the structural proteins of env and gag genes as well as additional viral polypeptides. In the control virions the major glycoprotein detected is gp71, interferon virions contain in addition to gp71 and 85k dalton (gp85) glucosamine-containing, fucose-deficient glycoprotein which is recognized by antiserum to MuLV but not by the gp71 antiserum. The surface iodination of the intact virions indicates that both gp71 and gp85 are the major components of the external virions envelope. However, unlike the control virions in which gp71 associates with p15E (gp90), the gp71-p15E complex was not detected in interferon virions. The analysis of the iodinated proteins of the disrupted interferon virions revealed the presence of 85k and 65k dalton polypeptides preciptable with antiserum against MuLV, which are not present in the control virions. The difference in the polypeptide pattern of virions produced in the presence of interferon does not seem to be a consequence of the slowdown in the synthesis of viral proteins or their processing in the interferon-treated cells. Both the structural proteins of env and gag genes seem to be synthesized and processed at a comparable rate in the interferon-treated and -untreated cells. These results indicate an alteration of virus assembly in the presence of interferon.


Evaluation of indapamide in the treatment of hypertension. A randomized double-blind, crossover trial was carried out in twenty hypertensive patients to evaluate the hypotensive efficacy and safety of indapamide. Indapamide in doses of 2.5 mg per day in combination with other antihypertensive agents was as effective in reducing the blood pressure in hypertensive patients as one tablet of Moduretic (50 mg hydrochlorothiazide and 5 mg amiloride). Indapamide induced no changes in serum potassium or uric acid levels and there were no major side effects.


Time- and dose-dependent changes in ejection fraction after anthracycline therapy. Cardiac toxicity due to anthracycline therapy is dose related, and congestive failure is a major limiting factor to therapy. Radionuclide cardiac evaluation provides a sensitive noninvasive method for detecting changes in cardiac function. Fifteen patients receiving either doxorubicin or daunomycin were evaluated with radionuclide ejection fractions (EFrn). The data indicate that the EFrn can detect an acute depressant cardiac action of these drugs as early as 24 hr after drug administration. In addition, a cumulative or chronic cardiac depression was noted; cumulative dosage of doxorubicin or daunomycin correlated with a reduced EFrn (p less than 0.001). We conclude that (1) the EFrn can noninvasively detect significant changes in cardiac function at low cumulative doses of doxorubicin or daunomycin and (2) the EFrn may be useful in evaluating cardiac function in patients during doxorubicin or daunomycin therapy.


Acute and chronic beta-receptor blockage with propranolol and the cardiovascular responses to intravenous sodium nitroprusside in the conscious rabbit. Sodium nitroprusside infused intravenously into conscious rabbits lowered blood pressure and raised heart rate and plasma noradrenaline. Acute beta blockade with propranolol, 2 mg/kg/hr i.v. for 2 hr, increased resting plasma noradrenaline and did not change the hypotensive effects of nitroprusside or the rise in plasma noradrenaline but attenuated the increase in heart rate. Chronic treatment with propranolol, 2 mg/kg twice daily for 3 weeks, did not modify the cardiovascular or reflex effects of nitroprusside. However, bilateral sinoaortic denervation augmented the hypotensive effect of nitroprusside and abolished both the heart rate and plasma noradrenaline responses to the vasodilator. The fall in blood pressure after nitroprusside is therefore attenuated by baroreflex mechanisms, which did not appear to be mediated by beta adrenoceptors or by withdrawal of vagal tone but could involve alpha-receptor-mediated vasoconstriction. Propranolol does not lower noradrenaline or modify the increase in noradrenaline after sodium nitroprusside.


Electrophysiological study of human ventricular heart muscle treated with quinidine: interaction with isoprenaline. We have investigated the effects of quinidine on the force of contraction and the intracellularly recorded action potential in papillary muscles isolated from human hearts. All preparations were obtained from patients undergoing corrective open heart surgery. The following results were obtained: (1) quinidine had a depressant effect on myocardial contractile force; (2) quinidine reduced the maximal upstroke velocity of the action potential; (3) quinidine shortened the plateau phase and prolonged the terminal repolarization of the action potential; (4) at higher concentrations quinidine reduced the resting potential; and (5) the depression by quinidine of both the plateau and the force of contraction was antagonized by isoprenaline. It is concluded that quinidine reduces the membrane conductances for sodium, calcium, and potassium ions. All of these actions of quinidine may contribute to the antiarrhythmic effects of the drug. The negative inotropic effect of quinidine can be explained by a depression of the calcium conductance at the myocardial cell membrane. The results show that earlier findings in laboratory animals regarding the effects of quinidine on the upstroke velocity and repolarization phase of the action potential are applicable to the human heart.


Electrophysiological effects of a new antiarrhythmic agent, flecainide, on the intact canine heart. The cardiac electrophysiology of flecainide a new antiarrhythmic agent, was studied in open-chested dogs. At plasma concentrations of 0.4 to 0.7 microgram/ml, flecainide significantly prolonged atrioventricular (AV) conduction. At plasma concentrations greater than 6.5 micrograms/ml, flecainide caused delay throughout the conduction system (77--240%). Low plasma levels of flecainide caused slight prolongation in atrial effective refractory period and AV nodal functional and relative refractory periods. At flecainide levels of 0.7 microgram/ml, ventricular response during atrial fibrillation was significantly slowed. The effects of flecainide on AV nodal and infranodal conduction and refractoriness were significantly enhanced by d,1-propranolol (0.5 mg/kg) but were unaffected by ouabain (0.01 mg/kg). The ventricular fibrillation threshold of supraventricular beats and ventricular premature beats increased with increasing plasma flecainide concentration (122% at 9.3 micrograms/ml and 172% at 1.3 micrograms/ml, respectively). Flecainide slowed ectopic atrial and ventricular pacemakers without affecting sinusor junctional pacemakers.


Pressor effect of beta-adrenergic receptor blocking drugs in spontaneously hypertensive rats treated with clonidine. The opposing effects on blood pressure of clonidine and beta-adrenoceptor antagonists which have been observed in some hypertensive patients were studied in conscious, spontaneously hypertensive rats. Daily treament with clonidine (100 micrograms/kg, given by gavage) for 2 weeks significantly reduced systolic blood pressure and heart rate. After 2 weeks of treatment with clonidine, administration by gavage of d,l-propranolol (10 mg/kg daily) or sotalol (10 mg/kg daily) or intraperitoneal injection of H 35/25 [d,1-erythro-4-methyl-alpha-(1-isopropylaminoethyl)-genzylalcohol hydrochloride] (10 mg/kg daily) concomitantly with clonidine significantly counteracted the hypotensive action of clonidine. d-Propranolol, metoprolol, and atenolol (all 10 mg/kg daily) had no such effect. All drugs except d-propranolol and H 35/25, a selective beta 2-adrenoceptor antagonist, enhanced the bradycardic action of clonidine. These findings suggest that the pressor effect of beta-adrenoceptor blocking agents during treatment with clonidine occurs at the level of beta 2-adrenoceptors in the periphery.


Effects of propranolol and timolol on calcium uptake by sarcoplasmic reticulum vesicles. The effects of the beta-adrenergic receptor blocking agents propranolol and timolol on the initial calcium uptake velocity of sarcoplasmic reticulum vesicles of rabbit skeletal muscle were studied. Racemic d- and l-propranolol had similar inhibitory effects on initial calcium uptake velocity, which was inhibited 50% by 5--7 X 10(-4) M racemic propranolol. Timolol was a much less potent inhibitor of initial calcim uptake velocity; 50% inhibition occurred at approximately 10(-2) M timolol. Both drugs inhibited maximal calcium uptake velocity; however, KCa (the Ca2+ concentration at which calcium uptake was half-maximal) was modified differently. Propranolol increased KCa, whereas timolol caused the KCa to decrease. Addition of either drug to an ongoing calcium uptake reaction at the time that calcium content became maximal caused renewed calcium uptake. The relative potencies of propranolol and timolol as negative inotropic agents are similar to their potencies as inhibitors of sarcoplasmic reticulum calcium uptake, but dissimilar to their beta-adrenergic receptor blocking potencies. Timolol, which has been reported to have less negative inotropic effect than propranolol, is approximately 5 time more potent than propranolol as a beta-adrenergic receptor blocking agent but 15 times less potent as an inhibitor of sarcoplasmic reticulum calcium uptake. Inhibition of sarcoplasmic reticulum calcium uptake may thus characterize negative inotropic potencies of new beta-adrenergic receptor blocking agents.


Nature of the central serotonin receptors mediating hypotension. The stimulation of central serotonin receptors leading to a decrease in blood pressure is well documented, but the nature of these receptors is not well known. It was therefore decided to characterize such receptors by assessing the modifications in blood pressure and heart rate after the intracerebroventricular administration of several drugs to anesthetized cats. Serotonin produced a dose-related decrease in blood pressure and heart rate, while parachloroamphetamine produced a slowly developing increase in blood pressure, and quipazine did not affect any of the parameters measured. The hypotension produced by serotonin was not modified by pretreatment with methysergide, was partially antagonized by quipazine, and was completely blocked by tolazoline. The present findings suggest that the central serotonin receptors which modulate blood pressure are different not only from peripheral receptors, but also from other receptors of this type.


Subsensitivity of cardiac beta-adrenoceptors in renal hypertensive rats. beta-Adrenoceptors were labeled with the selective beta-antagonist (-)[3H]dihydroalprenolol ([3H]DHA). Cardiac membranes isolated from renal hypertensive rats had the same density of adrenoceptors as normotensive rats (28 fmoles/mg of protein in both groups) but showed a significant elevation of the dissociation constant for [3H]DHA (Kd = 1.86 nM versus 1.04 nM for controls), indicating a reduced affinity of cardiac adrenoceptors for the radioligant. The lowered sensitivity to catecholamine is probably due to chronic exposure of beta-receptors to a high concentration of noradrenaline, whose turnover in cardiac nerves of renal hypertensive rats was significantly accelerated (turnover time 10.5 hr for hypertensive versus 17.2 for normotensive).


Antiarrhythmic and hemodynamic actions of flecainide acetate (R-818) in the ischemic porcine heart. The antiarrhythmic activity of flecainide acetate (R-818), 2 mg/kg, was investigated in anesthetized, open-chest pigs. Ventricular arrhythmias were provoked by reducing the flow in the left anterior descending coronary artery (LAD) to 25% of control during 30 min. During this period ventricular fibrillation occurred in 33% (11 out of 33) of control animals against 12.5% (1 out of 8) of animals treated with flecainide. Ventricular tachycardias were seen in 42% (14 out of 33) of the untreated animals as compared to none of the animals previously treated with flecainide. Total number of ventricular arrhythmias was significantly lower in the treated than in the untreated animals (p less than 0.05). However, when the LAD was occluded completely at its distal part, ventricular fibrillation occurred in all animals (5 untreated and 6 pretreated with flecainide). Time to onset of  ventricular fibrillation was the same for both groups of animals, despite a lower incidence of preceding ventricular arrhythmias in the pretreated group. Flecainide depressed myocardial contractility (LVdP/dt max), caused hypotension, and increased QRS width. Both myocardial depression and widening of QRS are related to arterial plasma levels of flecainide. Therefore, a slower infusion rate than the 1 mg/kg per minute used in this study is advisable when myocardial function is impaired.


Studies on the role of vasopressin in blood pressure control of spontaneously hypertensive rats with established hypertension (SHR, stroke-prone strain). We investigated the role of arginine-vasopressin (AVP) in maintaining the blood pressure of spontaneously hypertensive (SH) rats (stroke-prone strain) with established hypertension (22--28 weeks of age). In comparison with normotensive Wistar Kyoto (WKY) rats, plasma AVP concentrations of SH rats with benign hypertension (BH) were elevated twofold and in rats with severe or malignant hypertension (S-MH), fourfold. The height of the blood pressure was quantitatively related to plasma AVP in both BH and S-MH rats, the overall correlation coefficient being 0.66 (p less than 0.001). The intravenous injection of a specific AVP antiserum into conscious and unrestrained rats lowered blood pressure in 4 BH rats by 48 +/- 14 mm Hg and in 4 S-MH rats by 78 +/- 10 mm Hg and had only a marginal effect in 4 normotensive WKY rats. Infusion of saralasin did not lower blood pressure in WKY and BH rats and reduced blood pressure in only 2 of 7 S-MH rats tetsted (by 15 and 20 mm Hg). During AVP infusion the blood pressure of SH rats increased more (p less than 0.001) and heart rate fell much less (p less than 0.001) than in WKY rats. It is concluded that in SH rats with established hypertension, plasma AVP plays an important role in the maintenance of high blood pressure, while the renin-angiotensin system plays a minor or no role.


Effects of labetalol on plasma renin, aldosterone, and catecholamines in hypertensive patients. We studied the effects of labetalol, an alpha- and beta-adrenoceptor antagonist, on maximum exercise heart rate and on plasma renin, aldosterone, noradrenaline, and adrenaline levels at rest and during exercise in hypertensive patients. The dose of labetalol was doubled weekly from 0.3 to 2.4 g per day. The maximum exercise heart rate fell significantly during labetalol treatment, and there was a significant correlation between exercise tachycardia and the dosage of labetalol. Plasma renin activity and aldosterone concentration at rest decreased during treatment with labetalol. The exercise-induced increase in plasma renin activity was reduced by labetalol. Labetalol did not cause any significant changes in plasma noradrenaline and adrenaline at rest or during exercise.


Interaction of imidazoline alpha-adrenergic receptor antagonists with histamine receptors. Because the alpha-adrenergic receptor antagonists phentolamine and tolazoline are similar in structure to histamine, it is possible that the vasodilatation caused by these drugs may be due in part to stimulation of histamine receptors. The vascular effects of these agents were studied in the hindquarters of rats and the gracilis muscle of dogs. To eliminate interruption of sympathetic vasoconstrictor tone as a mechanism of vasodilatation, all animals were treated with the alpha-adrenergic receptor antagonist, dibozane. After dibozane, histamine caused vasodilatation in the rat, whereas both tolazoline and phentolamine caused vasoconstriction. It is concluded that phentolamine and tolazoline do not stimulate vascular histamine receptors in the rat. In the dog, after alpha-receptor blockade, phentolamine and tolazoline caused vasodilatation, as did histamine. Responses to histamine were partially attenuated by mepyramine and greatly attenuated by the combination of mepyramine and metiamide, indicating the participation of both H1- and H2-histamine receptors. Vasodilatation caused by phentolamine was not reduced by antihistamines and does not appear to involve histamine receptors. Vasodilatation following tolazoline was blocked by metiamide but not mepyramine. It is concluded that in addition to blockade of alpha-adrenergic receptors, tolazoline can cause vasodilatation by stimulation of histamine H2-receptors.


Prazosin first-pass metabolism and hepatic extraction in the dog. The short half-life, low plasma concentrations, and extensive biotransformation of prazosin suggest that it might be subject to extensive first-pass metabolism. Bioavailability, disposition, and hepatic extraction were studied in the dog. In conscious dogs whole blood prazosin concentrations were measured after oral and intravenous administration of the drug. Anesthetized dogs were used to measure prazosin concentrations in arterial and hepatic venous blood samples drawn simultaneously. The bioavailability of prazosin was 0.38 +/- 0.11. In anesthetized dogs the hepatic extraction of prazosin was 0.47 +/- 0.08 for a predicted availability of 0.53 +/- 0.08. Pharmacokinetic parameters were similar in conscious and anesthetized animals. Following intravenous administration to conscious dogs, prazosin concentrations in whole blood declined with a fast half-life of 3.9 +/- 1.74 min and a slow half-life of 153 +/- 24 min, the volume of distribution at steady state being 48.6 +/- 15.3 liters in dogs (mean weight, 22.6 kg). We conclude that prazosin availability following oral administration is low and that first-pass hepatic metabolism is largely responsible for this. A one-compartment model adequately describes prazosin pharmacokinetics in the dog.


Chronotropic effects of atropine sulfate and methscopolamine bromide in normal subjects and patients undergoing cardiac catheterization. The increase in heart rate caused by intravenous injections of atropine sulfate (AS) and methscopolamine bromide (MSB) were compared in 12 healthy volunteers. AS, 200 micrograms, or MSB, 100 micrograms, were injected every 3 min for five doses on successive days. Mean heart rate from the second and fifth injections of MSB was significantly greater than after corresponding injections of atropine. The ratio of equivalent doses of MSB and AS was found to be 1:3. Three subjects with AS and four subjects with MSB exhibited reduced heart rates after the first injection. Excellent correlation of plasma atropine concentrations and heart rate increase were obtained in two volunteers (correlation coefficient, r = 0.84). In the second study, AS, 180 micrograms, and MSB, 60 micrograms, were compared in 20 patients undergoing cardiac catheterization and coronary arteriography. Both drugs produced equivalent increments in heart rate. MSB produced a more predictable dose response than AS. Side effects were similar with both drugs when equivalent doses were used. This study confirms previous investigations that MSB can be titrated by intravenous injections to increase heart rate to desired levels. Absence of central nervous system stimulation by MSB suggests that it may be more appropriate for use in patients with myocardial infarction.


Effects of two monocarboxylic ionophores, grisorixin and alborixin, on cardiovascular function and plasma cation concentrations in the anesthetized dog. Grisorixin and alborixin are two monocarboxylic ionophores of the nigericin group which carry potassium preferentially over sodium. We studied their cardiovascular effects in anesthetized dogs. Grisorixin (2 mg/kg) and alborixin (1 mg/kg) produced a dramatic increase in ventricular contractile force (+250 and +320%, respectively) and in systolic and diastolic arterial pressures. At the same time, they induced a marked increase in plasma potassium in the coronary sinus and hepatic vein and a lesser increase in the renal and deep femoral veins. The concomitant modifications of the electrocardiogram were similar to those usually described in hyperkalemia, reflecting the effect of grisorixin and alborixin on plasma potassium. The parallel decrease in plasma sodium in the coronary sinus was more marked with alborixin than with grisorixin. These results suggest that the ionophores alter the intracellular ionic state of the myocardium, which may partly explain their positive inotropic effect and the greater effect of alborixin.


Regional myocardial perfusion and wall thickness and arteriovenous shunting after ergotamine administration to pigs with a fixed coronary stenosis. The hemodynamic effects of antimigraine drug ergotamine, which is considered contraindicated in patients with coronary artery disease, were studied in pigs with a normal myocardial circulation (doses of 8, 16, and 32 micrograms/kg, i.v.) or with acute coronary stenosis (8 micrograms/kg). In both groups of animals, ergotamine decreased heart rate, cardiac output, and arteriovenous anastomotic blood flow while increasing aortic blood pressure and systemic vascular resistance. No effects on total ventricular blood flow and its distribution within the myocardium were found in normal animals. In animals with a clamp on the left anterior descending coronary artery (LAD), the blood flow to the LAD-perfused area was reduced from 1.10 +/- 0.16 to 0.67 +/- 0.05 cm3/min/g. The endocardium was affected more than the epicardium and the endo/epi flow ratio decreased from 1.18 +/- 0.05 to 0.74 +/- 0.07. Ergotamine increased the blood flow to the ischemic zone towards normal values, and the endo/epi flow ratio to 1.05 +/- 0.21. However, myocardial wall thickness parameters, which showed functional deterioration during ischemia, did not change after ergotamine. The present study provides no clear support for cardiovascular contraindications to ergotamine administration.


Long-term antihypertensive therapy with prazosin plus a diuretic. Prazosin was administered to 21 diuretic-treated mild hypertensives for intervals of 1 month to 1 year. Three patients dropped out of the study, one from side-effects, one from noncompliance and the third for relocation. At 1, 6, and 12 months of follow-up therapy, blood pressures were reduced by 9--12%. Pulse rate was essentially unchanged. Thus prazosin appears to lower blood pressure for periods up to 1 year with little evidence of tolerance and with no change in heart rate.


Comparison of prazosin and methyldopa in essential hypertension: results of a randomized, double-blind, parallel trial. The value of prazosin and methyldopa administration was compared in 16 patients with essential hypertension during a 6 month randomized, double-blind, parallel trial. Both drugs were administered in combination with hydrochlorothiazide, 50 mg twice daily, until recumbent diastolic blood pressure was decreased to less than 90 mm Hg or the patient was receiving prazosin, 5 mg q.i.d., or methylodpa, 500 mg q.i.d. Addition of either prazosin or methyldopa to hydrochlorothiazide lowered blood pressure significantly and to a similar degree without altering plasma renin activity. Heart rate and weight were significantly increased during prazosin, but not methyldopa, therapy. Symptomatic side effects occurred with equal frequency during treatment with both agents and did not result in alteration or discontinuation of therapy in any patient. A decrease in frequency of administration from q.i.d. to b.i.d. or t.i.d./b.i.d. to q.d. did not adversely affect blood pressure control. Substitution of propranolol for prazosin or methyldopa provided similar antihypertensive effectiveness with a significant decrease in heart rate.


Renal perfusion changes during treatment of essential hypertension: prazosin versus propranolol. To evaluate renal hemodynamic changes during therapy of essential hypertension, we treated 12 essential hypertensives in crossover fashion, with 1 month each of placebo, prazosin, and propranolol alone. During prazosin treatment, blood pressure was normalized with preservation of glomerular filtration rate, renal plasma flow, and renal blood flow. Propranolol, however, normalized blood pressure with an associated significant decline in glomerular filtration rate, renal plasma flow, and renal blood flow. The potential mechanisms are discussed.


Effects of prazosin in the control of blood pressure in hypertensive dialysis patients. Twenty-two chronic hemodialysis patients with hypertension were treated with prazosin. Eight patients had volume-responsive hypertension, 11 volume-indpendent, and 3 high-renin hypertension. Blood pressure fell in all volume-responsive patients from a predialysis level of 175 +/- 5/100 +/- 3 to 148 +/- 4/75 +/- 3 mm Hg (p less than 0.001) after 3 months of therapy. Prazosin alone was effective in volume responsive patients at a dose of 5 +/- 1.0 mg daily. The blood pressure fell in volume-indpendent patients from 192 +/- 7/105 +/- 2 mm Hg predialysis to 155 +/- 6/80 +/- 3 after 3 months (p less than 0.001). Two were controlled on prazosin alone at a dose of 12 +/- 2 mg daily. Nine required 27 +/- 5 mg of prazosin daily as well as additional antihypertensive treatment. The blood pressure fell from 183 +/- 3/109 +/- 6 mm Hg predialysis to 173 +/- 17/85 +/- 3 mm Hg in high-renin patients after 3 months. One patient was controlled on 40 mg of prazosin daily. Two required 40 mg of prazosin daily as well as additional antihypertensive medication. Eleven patients described transient dizziness within the first month of therapy. One patient had recurrent syncope necessitating prazosin withdrawal; Prazosin is an effective antihypertensive agent which can be used in all types of hypertensive dialysis patients either alone or in combination with minimal side effects.


Isolation and characterization of free and membrane-bound polyribosomes from rabbit spinal cord. A procedure is described for the preparation of free and membrane-bound polyribosomes from rabbit spinal cord. First, myelin and filaments are floated away from other subcellular components. The latter are then fractionated by differential centrifugation followed by sedimentation through a discontinuous sucrose gradient. The ribosomal fractions are characterized by their electron microscopic appearance, RNA/protein ratios and sedimentation profile in a linear sucrose gradient. Both membrane-bound and free polyribosomes are active in incorporating amino acids in a cell-free system, whether heterologous or homologous pH 5 enzyme fractions are employed. Autoradiographic analysis of translation products separated by electrophoresis on SDS-polyacrylamide gels shows that both ribosome fractions are active in the synthesis of a variety of proteins including polypeptides which comigrate with alpha- and beta-tubulins and actin. The utility of these polyribosome preparations for the cell-free study of protein biosynthesis is indicated by the high molecular weight of many of the translation products, and by the similarity of the electrophoretic pattern of translation products from the cell-free systems to the pattern of radioactive protein synthesized by rabbit spinal cord during the hour prior to sacrifice.


Prazosin in hypertension with heart failure. Prazosin, in daily doses of 6--16 mg, was used along with furosemide and polythiazide in 11 hypertensive heart failure patients. Average supine and standing blood pressures were reduced from 180/108 and 171/106 to 130/84 and 135/86, respectively. Average heart rate decreased from 83 to 73 supine and from 84 to 75 standing. Changes in the indices of cardiac function--indlucing the ejection fraction, ejection time, and apical indices--each indicated improvement of left ventricular function. Thus prazosin along with diuretic agents may be particularly useful in the treatment simultaneously of heart failure and of elevated blood pressure with few side effects.


[The plasma concentration of beta 2-microglobulin in the diagnosis of malignancy of the gastrointestinal tract (author's transl)]. In patients with various stages of carcinoma of the colon-rectum (n = 42) or stomach (n = 15) the plasma concentration of beta 2-microglobulin was determined. The upper limit for normal was evaluated in a control group of healthy people (n = 36) and was found to be 2.4 mg/l. 36% of the patients with carcinoma of the colon-rectum and 27% of those with carcinoma of the stomach had higher than normal beta 2-microglobulin values. In the group of colon carcinoma patients there was a positive correlation between the extent of the tumor and the beta 2-microglobulin concentration. Thus, an appreciable frequency of increased values (greater than 50%) was found only in advanced stage carcinoma. In patients with carcinoma of the stomach only occasionally increased values were observed, independent of the stage of the tumor. In conclusion, the plasma concentration of beta 2-microglobulin is no adjunct in the early diagnosis of carcinoma of the gastrointestinal tract.


[Cross reactions of eight yeasts and their importance in serological Candida diagnostic (author's transl)]. While the antigen structure of yeast cell walls is well known, the immunological cross reactivity of these is often difficult to interpret. The cross reactions of eight yeasts were tested in rabbit hyperimmune sera by the indirect immunofluorescence test. Among the eight species examined were the medically important yeasts C. albicans, C. pseudotropicalis, C. krusei, C. parapsilosis, C. tropicalis, T. glabrata, and C. guilliermondii and the apathogenic yeast S. cerevisiae. It has been shown, that there is no correlation between the number of common antigens and the degree of cross reactivity which give identical titers. Therefore it appears, that the quantitative contributions of the single antigens have an important rule, i.e., some antigens are immunogenic in one species while having only hapten characteristics in another. Further the results show that in infections with C. krusei, C. pseudotropicalis, and C. guilliermondii homologous antigens have to be used for the serological diagnosis. The testing of human sera with the yeast antigens by the same methods showed that the strikingly distributions of the Candida titer and the Saccharomyces titer were similar. A larger number of sera of healthy people and patients were compared in the hemagglutination test. These results showed that pathological C. albicans titers cannot be caused by S, cerevisiae antibodies.


Identification of Bacteroides fragilis by indirect immunofluorescence. Rabbit antiserum against live or heat-killed Bacteroides fragilis NCTC 9343 bacteria was titrated against hot phenol water-extracted polysaccharide antigens from five different species of the 'B. fragilis group' of bacteria using an enzyme immunoassay and shown to be specific for the B. fragilis NCTC 9343 polysaccharide. When the antiserum was used in indirect immunofluorescence, 97.1% of 244 B. fragilis strains were correctly identified. Only 8 of the other 312 Bacteroides strains were stained by the anti-B fragilis serum. The 'cross-reactive' Bacteriodes strains all belonged to the "B. fragilis group" of bacteria (i.e., B; distasonis, B. ovatus, B. thetaiotaomicron, B. uniformis, and B. vulgatus). None of the 425 aerobic enteric bacteria representing Salmonella, E. coli, Proteus, Yersinia, Shigella, Klebsiella, Enterobacter, Citrobacter, and Pseudomonas were positive using the anti-B. fragilis serum. Likewise, all the 59 gram-positive strains representing Streptococcus, Bacillus, Peptostreptococcus, Peptococcus, Propionibacterium, Lactobacillus, Eubacterium, and Clostridium did not stain. Our data shows in accordance with other findings [11], that B. fragilis strains possess a species-specific cell envelope antigen(s) which promises to be important for production of antisera, making a rapid identification of the species possible.


Neutral protease activity in lymphocytes of Lewis rats with acute experimental allergic encephalomyelitis. Lymphocytes from popliteal and inguinal lymph nodes of Lewis rats with acute EAE as a result of injection of lyophilized guinea pig myelin in Freund's complete adjuvant exerted strong proteolytic activity at neutral pH toward myelin basic protein. After injection of myelin the level of proteolytic activity remained about the same as that in lymphocytes from Freund's adjuvant-injected controls until about day 10 after injection, just before the onset of paralytic symptoms; then the proteolytic activity increased to approximately double its former level. Myelin basic protein was hydrolyzed by whole lymphocytes, but more activity was unmasked by homogenization. Similar results were also obtained using lymphocytes from thymus of EAE and control animals. Lymphocytes with high levels of proteolytic activity were not absorbed by glass wool, did not stain with neutral red, nor did they phagocytose antibody-coated sheep red blood cells. Thymus and lymph node lymphocytes cleaved myelin basic protein to three major peptides and a fourth minor peptide, while spleen lymphocytes hydrolyzed basic protein at only one point resulting in two peptides whose molecular weights added up to that of myelin basic protein. The protease activity was inhibited by 5 X 10(-3) M p-chloromercuribenzoate and by phenylmethyl sulfonyl fluoride, TPCK, and soybean trypsin inhibitor, therefore the enzymatic activity probably depends on a serine residue and a sulfhydryl group. The bulk of the enzymatic activity is mostly membrane bound with the highest specific activity and total activity contained in a lysosomal-mitochondrial fraction. In view of the infiltration of lymphocytes into the brain substance in acute EAE, it is suggested that these cells may contribute to the destruction of myelin which is usually attributed to the monocyte or macrophage.


Persistence of nucleolar RNA-rich structures and Ph1 duplication in the blastic crisis of chronic myeloid leukaemia. Nucleolar persistence in metaphase plates is a feature observed in most of the cells in neoplastic processes. Pathological persistence or fragmentation of the nucleoli is thought to be the cause of some numerical chromosomal aberrations due to non-disjunction of the chromatids, with particular involvement of the satellite chromosomes. Thus, a combined selective staining of both the nucleoli (amido black 10B according to Mundkur and Brauer's cytochemical technique) and the chromosomes (neutral red) was applied to the metaphase plates of patients with chronic myeloid leukaemia in the blastic crisis. Duplicated Ph1 was associated with amido black-stained areas at a very high rate in some cases. Since the blastic crisis in chronic myeloid leukaemia is characterized by the appearance of an increased number of immature, highly nucleolated cells, these findings lend support to the hypothesis that the duplication of the Ph1 represents a feature possibly favoured by the pathological persistence of nucleolar RNA-rich structures in the metaphase.


APUD cells in normal and hyperplastic prostates. Argyrophilia and argentaffinity, as basic properties of APUD cells, were investigated in 50 normal and hyperplastic prostates, which included both autopsy and surgical specimens from patients of various ages. Normal prostates (including glands from 3 foetuses) had 62% of argyrophil-positive granules in the glandular epithelia, while only 44% of the hyperplastic glands were positive. Argentaffin-positive cells were found in 12% of the surgical hyperplastic cases. Both argyrophil and argentaffin cells were distributed in zones, often in lobule-like shapes, lying along the basal membrane. On the basis of these findings, there is a discussion on the possible roles played by the so-called APUD cells in hyperplastic and neoplastic growths of the prostate, such as carcinoid tumours (apudomas) or endocrine-associated syndromes in the course of prostatic cancer.


Sharing Ia antigens between species. III. Ia specificities shared between mice and human beings. Certain mouse alloantisera have been found to detect immunologic cross-reactions between human and murine Ia antigens. Almost every anti-Iaa, -Iak and -Iad serum tested exhibited such cross-reactions. Sera prepared against the products of limited segments of the mouse I region and tested on human B cells revealed that anti-I-E/Ck cross-reactions were more readily detectable than anti-I-A, B, Jk cross-reactions. Most of the mouse alloantisera were cytotoxic to bells from almost every individual tested, although a few sera exhibited more restricted patterns of lysis, permitting limited segregation analyses. The cytotoxicity of these mouse alloantisera in family studies was consistent with HLA linkage of the genes responsible for the cross-reacting Ia determinants. Immunochemical analysis on radiolabelled detergent lysates of human lymphocytes indicated that the sera reacted with molecules of 34,000 and 28,000 daltons. Thus, by cellular distribution, HLA association, and immunochemical criteria, these mouse alloantisera detect human Ia antigens. These cross-reactive sera should be of practical value for the detection of human Ia antigens and may also have theoretical implications for the evolution of genes coding for Ia antigens.


Clearance and glomerular localization of preformed DNP anti-DNP immune complexes. The in vivo behaviour of well-defined immune complexes in rats was studied using complexes derived from DNP-conjugated bovine thyroglobulin (DNP-BTG) and purified specific goat anti-DNP IgG. Both clearance and glomerular localization were mainly dependent on the nature of the antigen. Soluble immune complexes formed with DNP17-BTG were cleared faster and showed a more marked localization in the glomerular mesangium than complexes formed with DNP3.4-BTG. A slight increase in the antibody to antigen ratio seemed to facilitate mesangial localization of soluble immune complexes. Insoluble immune complexes showed temporary localization as microemboli in the lumina of glomerular and peritubular capillaries. This study thus shows that not only the size and composition of the complexes but also the nature of the antigen within the complex can influence the clearance and organ localization of circulating immune complexes.


T cell independent induction of antigen specific suppression of the antibody response. Immune spleen cells (from mice given 2 x 10(7) HRBC 14 days earlier) when mixed in vitro with carrier-primed syngeneic spleen cells (from mice given 2 x 10(5) HRBC 3 days earlier) are able to suppress the anti-TNP and anti-HRBC PFC response to TNP-HRBC. If immune thymocytes are substituted for spleen cells suppression is not observed. This suppression is antigen specific, resistant to anti-T treatment or x-irradiation, and is exerted by nylon wool-retained cells of the immune spleen cell population. An antigen specific suppressive factor is released from immune spleen cells in culture. Under these experimental conditions, suppression appears to be mediated by a specific product of B rather than T cells present in the immune spleen cell population.


A thymus-independent IgG response against dextran B512 can be induced in C57BL but not in CBA mice, even though both strains possess a VHdex gene. CBA and C57BL mice both possess a VHdex gene coding for antibodies against the alpha 1-6 epitope of dextran B512. After immunization with dextran, CBA mice produce IgM plaque-forming cells (PFC) only, which show regular cyclical fluctuations. The second PFC peak disappeared after injection of dextranase, indicating that it is antigen-dependent. The anti-dextran response in C57BL mice is characterized by only one IgM peak, followed 1 day later by an IgG peak that may exceed the IgM response by a factor of 10. The IgG anti-dextran response in C57BL mice was thymus-independent. CBA mice gave an IgG response to the hapten fluorescein isothiocyanate (FITC) after immunization with FITC-dextran, indicating that dextran can function as a carrier for an IgG response in this strain. Attempts to induce IgG PFC against dextran by immunizing CBA mice with thymus-dependent dextran-protein conjugates consistently failed, althouth the conjugates induced IgG fc in C57BL mice. Spleen cells from CBA mice failed to produce IgG antibodies against dextran after transfer into lethally irradiated C57BL mice, whereas the C57BL spleen cells produced IgG PFC after transfer into CBA mice. The lack of IgG synthesis against the alpha 1-6 epitope of dextran in CBA mice appears to be regulated exclusively at the B cell level and is restricted specifically to the VHdex gene product.


The effect of dextran on the incidence of thrombosis in microvenous Nakayama ring pin anastomoses. Including further studies on the histopathology of Nakayama anastomoses. Anastomoses of the posterior facial veins in the rabbits neck (diameter appr. 1.5-2.0 mm) were performed under the operating microscope, using the Nakayama ring pin stapler. Immediately before the operation the animals received clinical dextran (mw 70000) or physiological saline intravenously in a double-blind randomized pattern. The total incidence of partial or complete thrombosis in this experiment was low (around 10%). No difference between the two groups were detected. All anastomoses were examined in vivo, under the microscope and histologically. The histological findings are penetrated.


Analysis of HGPRT- CRM+ human lymphoblast mutants. Three 6-thioguanine-resistant mutants of the human diploid lymphoblast line MGL-8 were studied. The inactivation by heat of both HGPRT activity and antigenicity of the HGPRT immunologically cross-reacting material of the A30 mutant cells were not protected by PRPP, indicating that the HGPRT in A30 cells has an altered PRPP binding site, leading to lack of stabilization and rapid degradation of the enzyme. Two dimensional separations of the immunoprecipitates from extracts of the parental and mutant cell lines showed that the A35 mutant CRM has a more acidic isoelectric pH, while the A30 CRM has a more basic isoelectric pH and that the A30 protein has a faster rate of degradation than the wild-type HGPRT. The A30 CRM also has a smaller molecular size than the wild-type enzyme.


Colorectal carcinoma antigens detected by hybridoma antibodies. Hybridoma cells which secrete colorectal carcinoma-specific antibodies have been produced and used to study the antigenic structure of these tumor cells. Nineteen antibodies have been studied in detail, and 15 of these are colorectal carcinoma specific. Only two antibodies reactive with carcinoembryonic antigen (CEA) have been discovered and five other antibodies that react with distinct epitopes on the cell surface have been defined. Several antigens with distinct molecular characteristics have been shown to exist by use of hybridoma antibodies. Six hybridoma antibodies have been shown to mediate antibody-dependent cell-mediated cytotoxicity (ADCC).


Regulation of expression of type C virion DNA polymerase (reverse transcriptase) in human x mouse and human x rat hybrid cells. Human cells from 13 different individuals were fused to mouse and rat cells producing abundant type C viral particles. The results demonstrate that incorporation of active DNA polymerase (reverse transcriptase) into mature type C particles is suppressed in many of the hybrid clones but not in the parental mouse cell clones. This low particle-associated DNA polymerase phenotype was a heritable trait for over 100 cell generations but reversion to a high particle-associated DNA polymerase phenotype was possible. In contrast, no evidence for suppression of viral p30 antigen was found. These results suggest that human cells contain a factor(s) capable of interfering with the normal maturation of the mouse retrovirus DNA polymerase protein; however, it was not possible to assign this function to any of 20 different human chromosomes tested. It is suggested that these somatic cell hybrids may be useful in examining individual events in retrovirus packaging and release.


HLA antigens in a psoriatic family: comparative studies with MLC, HTC, PLT and serological HLA-DR determinations. HLA genotypes were characterized in a large family of 48 individuals in three generations. In this family, carriers of the proband's disease-predisposing haplotype commonly expressed clinical signs of illness, manifested as psoriasis and/or arthritic lesions. From these data, and from other family studies, presented previously, we have concluded that cutaneous and/or joint lesions may be signs of disease in carriers of the predisposing HLA haplotype. We have investigated HLA-A, B, C and D/DR antigens in the family members. This gave us the opportunity to evaluate the validity of different assays for the determination of HLA-D alleles in a family material where the MLC tests formed the basis for a correct assignment of HLA-D determinants. The HTC method and the PLT assay were both afflicted with specific typing problems, partly due to the existence of cellular cross-reactions between HLA-D determinants, which seemed to be reminiscent of serological DR crossreactions.


Two HLA-D and DR alleles are associated with coeliac disease. A group of 45 children affected with Coeliac Disease (CD) was typed for HLA-A, B, C, D, and DR specificities. The most significant associations were found with two alleles of the D series, with both cellular and serological typing. It is suggested that the susceptibility to CD is determined by two different genes within the HLA region, the first in common with organ-specific autoimmune diseases and associated with DW3, the second possibly specific for CD and associated with Dw7.


Improved B cell typing for HLA-DR using nylon wool column enriched B lymphocyte preparations. A rapid, simple procedure for preparing B lymphocytes using nylon wool has been devised. When mixtures of T lymphocytes, B lymphocytes, and monocytes are applied to nylon wool columns, B lymphocytes can then be removed, virtually free of monocytes. Such preparations (Ad) are 85 +/- 6% SIg+ and have only 3 +/- 1% monocyte contamination. The yield from peripheral blood averages 5% of all mononuclear cells. In contrast, the E rosette depletion method (E-) yields cells which are only 64 +/- 5% SIg+ and have 25 +/- 7% monocyte contamination. The superiority of the nylon method for HLA-DR typing was demonstrated in a comparative study of Ad and E- with eight normal individuals. Cytotoxic scores were higher and a large number of reactions, representing DRw groups and additional cross-reactions, was detected.


Influence of genes associated with the main histocompatibility complex on deoxyribonucleic acid excision repair capacity and bleomycin sensitivity in mouse lymphocytes. In sets of mice congenic at H-2 and upon two backgrounds, and selected according to known differences in strain-specific lifespans, DNA repair efficiency in spleen cells was compared by two techniques: excision repair capacity following UV-irradiation, and bleomycin sensitivity. Significant differences between certain congenic partner sets were noted with both techniques, suggesting that the main histocompatibility complex influences DNA repair capacity.


HLA frequencies in a Mexican Mestizo population. The aim of the present study was to reevaluate the distribution of HLA antigens in Mexican Mestizos since, in our previous report, very few specificities were explored for this population. The Mestizos are primarly a mixture of Caucasians (Spaniards) and Mexican Indians and account at the present time for about 95% of the total Mexican population. A and B antigens were typed on isolated cells using a microlymphocytotoxicity technique. Antigen, gene and haplotype frequencies were calculated including the significance for delta values. The results clearly showed that the HLA distribution differs from other ethnic groups and the predominant antigens are A2, A9, B5, Bw35 and B40, but the general pattern clearly shows the participation in genetical composition of Spanish and Mexican Indian backgrounds. The most frequent haplotypes were A2-B5, A9-Bw35, A9-B40 and A2-B40, which are also the most common in some Mongoloid populations.


False negative HLA-DR assignment necrokidney donors. A glucocorticoid effect? Thirteen potential necrokidney donors were HLA-DR typed using peripheral blood B lymphocytes (PBBL). Antigen assignment was impossible in 10 cases because of non-reactivity of the PBBL with the HLA-DR testserum set, in some of the cases in spite of positive reactions with a positive control xenogeneic anti-human B cell antiserum. In five of these 10 donors, HLA-DR typing was also performed on B lymphocytes isolated from the spleens, and in all five cases strong specific reactions were obtained, allowing reliable antigen assignment. Evidence is provided that the negative serum reactions with the five donors' PBBL were "falsely negative". It is suggested that glucocorticoid treatment prior to blood sampling for HLA-DR typing may be responsible for the phenomenon.


Androgen receptors in benign and malignant prostatic tissue. A new procedure has been developed to purify prostatic cell nuclei and to achieve a high yield of nuclear material for study. This technique allowed the investigation of the enzyme 5-alpha-reductase and its relationship to the translocation of dihydrotestosterone and androgen receptors into the nucleus of the prostatic cell. The studies revealed that 5-alpha-reductase was localized chiefly to the stromal components of all types of prostatic tissue. Although there is a higher concentration of dihydrotestosterone in carcinoma tissue the 5-alpha-reductase activity is low. Studies on the effect of androgens and antiandrogens on nuclear uptake of androgen revealed that androgens are potent stimulants of receptor translocation, while antiandrogens stimulate little or no translocation.


Keratinization of the conjunctiva.

Keratinization of the conjunctiva. Keratinization of the tarsal conjunctiva in an eye with adequate tears occurs following a number of conditions, including irritation and sensitivity to topical medications, Stevens-Johnson syndrome, radiation to the lid, and occasionally from unknown causes. The keratinized cells produce an epithelial keratitis with subsequent vascularization of the cornea. The morphogenesis of this condition is discussed, along with therapy including mucous membrane grafts and freezing of the tissue.


[Characteristics of immunoglobulin dynamics and absorption in newborn calves]. A study on protein dynamics in calf blood serum from the time of birth to age of 60 days was carried out by use of electrophoresis on 1% agar gel, immunodiffusion and immunoelectrophoresis with rabbit immune serum against cattle blood serum, colostrum whey and pure nu-globulin. Investigations were performed on the 4th and 24th hour, and on the 3rd, 6th, 8th, 10th, 15th, 20th, 30th, 45th and 60th day after birth. Resorption of colostral gamma globulins, applied intravenously and orally in the form of the preparation lactoplasmin and 10% normal gamma-globulin injected intravenously was followed under experimental conditions. It was established that calves are born with a comparatively low protein level in the blood serum and after feeding with colostrum begins, specific quantitative and qualitative changes in their profile take place, but the changes concerning gamma-globulins are most pronounced. gamma-globulins are best resorbed after oral application of colostral globulin preparations, and not so well after intravenous application. Noral blood gamma-globulin injected intravenously is least resorbed. Eventual causes for this fact are discussed.


The development of an experimental model of subretinal neovascularization in disciform macular degeneration. A reproducible model of subretinal neovascularization has been developed, an important first step in the study of disciform macular degeneration. The methodology and clinical description of subretinal neovascularization in the primate have been provided and histopathologic and electron microscopic correlations included. This model provides a most promising and useful mechanism for the evaluation of the many different factors and hypotheses that have been proposed for the genesis of the disciform response. In humans, it probably has more relevance to the traumatically induced disciform process, particularly that after laser therapy, than to the senile degenerative process. The purpose of this investigation is to develop the animal model by which many different theories regarding the pathogenesis of the disciform response can be tested. These initial studies relate to ischemia, hemorrhage, the inflammatory response, and position in relation to the center of the vascular-free zone as factors important in the development and evolution of subretinal neovascularization.


Effect of nutrients on the toxicity of pesticides carbofuran and hexachlorocyclohexane to blue-green alga Nostoc muscorum. The effects of various levels of nutrients like potassium phosphate (dibasic), calcium nitrate, and calcium chloride individually and in combinations were studied on the toxicity of the commonly used pesticides carbofuran and hexachlorocyclohexane (HCH) in growth medium to the N2-fixing blue-green alga Nostoc muscorum. It was observed that all these chemicals had some effects on toxicity. The toxicity of both carbofuran and HCH was reduced to some extent in the presence of higher concentrations of the nutrients when compared to normal levels of the chemicals in the medium. The higher doses of nutrients in combinations antagonized the effect of individual treatment and enhanced the toxicity of pesticides.


[Host range relationship of bacteriophages T3 and T7 with Escherichia coli K 12 strains]. The closely related phages T3 and T7 exhibit different growth patterns on Escherichia coli W hosts cells (E. coli K12 derivatives). T7 grows normally while T3 does not adsorb. T3hw mutants displaying a T7-like host range were isolated and described.


An in vitro culture system for crayfish organs. Organs of crayfishes--mainly integument of Astacus leptodactylus--have been kept in an in vitro culture of several days. The medium consists of 2 parts of van Harreveld saline and one part of horse serum, supplemented with antibiotics for long term cultures. This medium is very similar to the Astacus hemolymph in its ionic content, the pH, the osmolarity, the protein concentration and the cholesterol content. The glucose concentration is somewhat higher and there are differences in the nitrogen excretory end products. Integument of Astacus is able to synthesize protein, RNA and DNA in vitro for several days. Ecdysone is converted to 20-OH-ecdysone by the integument under in vitro conditions.


Observations on the chromatin staining by aluminium-hematoxylin. The staining reaction of chromatin by aluminium-hematoxylin was investigated. Intense staining, which proved dependent on nucleic acid content, was achieved by using either the preformed lake, mordanting followed by hematoxylin, hematoxylin alone or the lake at high ionic strength. Extraction procedures removed aluminium ions from chromatin but the lake was retained. Chromatin fluorescence induced by ethidium bromide and acridine orange was abolished after staining with the lake. Intercalating-like forces are suggested for aluminium-hematoxylin binding to chromatin.


Effect of DDT and gamma-HCH on the biological properties of staphylococci. The effect of DDT and gamma-HCH in concentrations of 100, 50, and 10 micrograms on the biological properties of staphylococci was studied. Both substances proved to be inhibitory to coagulation of rabbit plasma and to penicillinase production, causing depigmentation of the colonies. The morphological structure of cells is affected by these substances as well. The growth of the strain is not substantially influenced. Gas chromatography proved that staphylococci do not participate in the DDT and gamma-HCH degradation in the medium.


[The antigenic relationship between Kloeckera and Salmonella cholerae-suis O antigen (author's transl)]. The immune sera for Kloeckera africana, K. corticis, K. javanica var. javanica and K. javanica var. lafarir agglutinated Salmonella cholerae-suis (O:6(2),7). The immune serum for S. cholerae-suis agglutinated K. africana, K. corticis, K. javanica var. javanica and K. javanica var. lafarii. Absorption and agglutination cross tests demonstrated common antigenic factor(s) between above mentioned yeasts and Salmonella O:7 antigen and also K. javanica var. lafarii and Salmonella O antigens 6 and 7.


[On the presence of 20-methylcholanthrene in the atmospheric air (author's transl)]. The strongly carcinogenic polycyclic aromatic hydrocarbon 20-Methylcholanthrene was detected in the urban atmospheric air in Budapest (the capital of Hungary). The main source of this pollution seems to be the motor vehicle traffic, since samples from a heavy traffic junction contained up to six times higher 20-Methylcholanthrene concentrations than samples from a low traffic area. Ultraviolet absorption spectrophotometry yielded similar results. The presence of 20-Methylcholanthrene is apparently connected with the presence of 3,4-Benzpyrene and 1,2-Benzpyrene, respectively.


Effect of cyproterone acetate on the genital organs of female rats. Effects of cyproterone acetate, a synthetic steroidal compound, on the reproductive organs of female rats have been investigated. This agent causes reduction in ovarian weights indicative of suppression of pituitary gonadotrophins. Oestrogenic nature of cyproterone acetate was investigated in intact and ovariectomized animals taking uterine weight, vaginal keratinization and other oestrogen sensitive biochemical parameters. Cyproterone acetate in ovariectomized animals induced vaginal keratinization, increase in uterine weight and uterine protein, RNA, glycogen and sialic acid contents. These effects were parallel to the effect of oestradiol dipropionate in ovariectomized animals, thus indicating oestrogenic activity cyproterone acetate. The histological and biochemical parameters lead to the conclusion that cyproterone acetate possessed antifertility property due to its inherent oestrogenic nature.


Correlation between hepatic transport of cholephilic organic anions and their effect on hepatic mitochondrial respiration. The maximum biliary excretion rate (Tm) of some nonmetabolized organic anions was investigated in anaesthetized male rats and was found to increase in the following order: indocyanine green (ICG), rose bengal (RB), bromsulphthalein (BSP; its conjugation with glutathione was prevented by diethyl maleate), bromcresol green (BCG), eosine (E), BSP-glutathione conjugate (BSP-GSH), iodoxamic acid (IA) and amaranth (A). The accumulation of ICG, RB, BSP and BCG in liver was relatively high, whereas E, BSP-GSH,IA and A were taken up by the liver in a lower concentration. ICG, RB, BSP and BCG decreased the biliary flow, however, E, BSP-GSH, IA and A stimulated it. ICG, RB and BSP had a strong, BCG and E a moderate, inhibitory effect on hepatic mitochondrial respiration stimulated by ADP, DNP and glutamate-malate, whereas BSP-GSH, IA and A had no effect on this process. Our results suggest that the depressing effect of cholephilic organic anions on bile production and their low biliary excretion rates are connected with their inhibitory effects on mitochondrial respiration.


Hepatic microsomal induction and hepatic transport. Hepatic microsomal induction (hexobarbital sleeping time, cytochrome P-450 and microsomal protein concentration, liver weight) and hepatic transport (hepatic uptake, biotransformation, biliary excretion) have been studied in rats. Phenobarbital pretreatment (75 mg/kg i.p. daily for 5 days) produced microsomal induction in the liver and enhanced biliary excretion of bromcresol green, eosine, bromsulphthaleine glutathione conjugate (BSP-GSH), amaranth and iodoxamic acid. However, biliary excretion of indocyanine green was unchanged after phenobarbital pretreatment. Biotransformation of bromsulphthalein (BSP) with glutathione was also increased by phenobarbital. The hepatic concentration of these organic anions was not influenced uniformly after phenobarbital pretreatment: the concentration of indocyanine green, bromcresol green, eosin and BSP-GSH in the liver was unchanged, that of amaranth and iodoxamic acid was enhanced following phenobarbital pretreatment. Investigation of the effect of pretreatment with other barbiturates showed that barbital, butobarbital, pentobarbital and amobarbital produced microsomal induction. Only baribtal and butobarbital stimulated biliary excretion of organic anions, whereas pentobarbital and amobarbital proved to be ineffective in this parameter. The results seem to indicate that the enhanced biliary excretion of exogenous organic anions produced by barbiturates is independent of microsomal enzyme induction.


A neurolinguistic longitudinal study of a pure motor aphasia. We have made a detailed neurolinguistic study of a patient with motor aphasia. The method used for his reeducation and the variants found in his language from a linguistic point of view through the course of several months are described. During this study we have found similarities with cases studied by Marcie reached by him in the errors common in patients with motor aphasia. But, contrary to Marcies, findings, we have found phonemes and allophones outside of the phonological and/or dilectal system of the language in this area. These phonemes and allophones are described together with the environment in which they appear.


HLA types in corneal diseases.

HLA types in corneal diseases. Evaluation of the results of HLA typing of 187 patients grouped into herpetic keratitis (37), non-herpetic keratitis (43), keratoconus (42), endothelial dystrophy (23), stromal dystrophy (13), lues (5), and injuries (24), failed to show convincing deviations in any of the groups from a normal control series (2900 persons). Yet, as for the herpes group, a rise in B5 must strongly be suspected. The data are presented and possible implications are discussed.


Corneal endothelial permeability following storage in moist chamber and MK medium. Rabbit corneal endothelial permeability to sucrose and dextran was determined after storage in moist chamber or MK medium. When compared to fresh rabbit corneas there was no statistically significant difference in endothelial permeability to these two solutes after both types of storage for periods up to ten days.


Hypertrophic response of the kidney in indomethacin treated rats. Indomethacin, a potent inhibitor of prostaglandin (PG) synthesis administered to rats in a total dose of 2 x 4.5 mg/kg i.v. for 2 days enhanced compensatory renal hypertrophy following unilateral nephrectomy (UN). The same drug given to rats with intact kidneys in a total dose of 4 x 4.5 mg/kg i.p. in 2 days increased kidney weight renal RNA content and decreased renal acidic ribonuclease activity. It remains to be elucidated whether the suppression of PG synthesis and/or other pharmacological effects of indomethacin account for these findings.