Effect of serum and blood on Enterobacteriaceae grown in the presence of subminimal inhibitory concentrations of ampicillin and mecillinam.

Bacteria from the strains of five species of Enterobacteriaceae grown in the presence of subminimal inhibitory concentrations of ampicillin or mecillinam formed into filamentous or round cells. These filamentous and round cells as well as normal control bacteria were incubated with either fresh human serum or blood, the bactericidal effects of which were then determined. In most cases, the bactericidal effect of either serum or blood on filamentous or round cells was less than the effect on control cells. In some cases, the effect on these drug-exposed cells was similar to that on control cells, but in no instance was the effect greater for the drug-exposed cells than for the control cells. However, in all cases in which the bactericidal effect of either serum or blood on the control cells was greater than or equal to 99%, the bactericidal effect on the drug-exposed cells was close to 90%. Although drug-exposed cells were not much more resistant to the bactericidal effect of serum or blood than were normal bacteria, they clearly were not more susceptible to these effects.


Drug-inactivating enzymes of bacteria grown in subminimal inhibitory concentrations of antibiotics.

Repeated transfers of strains of Escherichia coli and Staphylococcus aureus in medium containing subminimal inhibitory concentrations (sub-MICs) of gentamicin caused a moderate increase in the minimal inhibitory concentration of gentamicin. At the end of such transfers, E. coli K12 produced aminoglycoside phosphotransferase(3')-I [APH(3')], AND E. coli R112, which carries the plasmid-coded enzyme APH(3')-I, also produced the acetylating enzyme aminoglycoside acetyltransferase(2') [AAC(2')]. E. coli R148, which produces aminoglycoside phosphotransferase(3')-II [APH(3')-II], did not change its output of enzymes. Repeated transfers to media containing increasing concentrations of gentamicin resulted in the development of complete resistance to all aminoglycosides without the concurrent development of any demonstrable new enzyme activity. With repeated transfers in drug-free medium, a complete reversal to susceptibility to gentamicin, but not to other aminoglycosides, was obtained for strains that had previously been transferred in sub-MICs of gentamicin, whereas strains that had been transferred in increasing concentrations of gentamicin did not revert to their original sensitivity to aminoglycosides despite repeated transfers in drug-free medium.


[Effectiveness of treatment with rifampicin and isoniazid in association with ethambutol or prothionamide in patients with recently detected pulmonary tuberculosis].

In newly discovered patients with cavitary forms of pulmonary tuberculosis treatment with rifampicin, in association with other drugs, leads to recovery (negativation) in 69,6 percent of the patients in 3 months, in 92 per cent after 6 months and in 94,6 percent after 9 months of treatment. Closure of cavities is found in 23,4 percent, 64,5 percent and 76,6 percent of all patients at the corresponding time intervals. Comparisons of the various regimens of chemotherapy which include rifampicin showed that the maximal efficiency was obtained with rifampicin, ioniazid and a third drug (either etambutol or protionamide). The time intervals for the disappearence of bacilli and for the closure of cavities depend to a great extent on the degree of development of the pathological process, on the type of the lesions and the intensity of bacilli elimination at the start of the treatment. The use of rifampicin in new patients, in association with other drugs, leads to a rapid negativation, a shortening of the time necessary for the closure of cavities, especially in patients in whom the disappearence of the bacilli from the sputum occurs in the first three months.


[Immunoglobulins in clear-fluid pleurisy (preliminary note)].

The study of IgG, IgM and IgA immunoglobulins in the pleural exsudate and in the serum of 34 patients with pleurisy of various etiologies shows an increase in the level of serum immunoglobulins as compared with the level in the exsudate. The only immunoglobulin fraction showing variations in relation with the etiology of the disease in the IgG fraction, which is increased in inflammatory exsudates, in contrast with those found in pulmonary cancers. Recovery without sequellae of pleurisy supposes the presence (in situ) of an increased amount of IgG. These observations are supported by a good statistical-mathematical correlation (p less than 0,01 and 0,001).


[Current aspects of recovery and invalidism in recurrent pulmonary tuberculosis].

An analysis was made, 12 months after strictly supervised treatment which included rifampicin and/or etambutol, of 83 cases of relapses of pulmonary tuberculosis. Therapeutic aspects were studied, as well as problems in relation to the possibility to continue work, as well as temporary invalidity. The therapeutic curing of the relapses was achieved in 74 patients (89,15 percent), while 9 remained with a positive bK in the sputum. The recovered subjects continued their activity without any changes in the working place, and retirement was accepted for active tuberculosis in 12,5 percent of the patients, while retirement for posttuberculous syndrome in another 7,14 percent of the patients. Males were predominant (81,81 percent of the total), mostly aged between 30 and 49 years, originating from rural environments.


A rapid method for the separation of functional lymphoid cell populations of human and animal origin on PVP-silica (Percoll) density gradients.

Subclasses of lymphocytes can be separated on gradients of non-toxic polyvinylpyrrolidone-coated colloidal silica (Percoll) by virtue of differential densities. Such gradients can yield functionally active lymphocyte populations after brief centrifugation. Gradients can be generated in a discontinuous step fashion and centrifuged in standard table-top laboratory centrifuges or as self-generating gradients during ultracentrifugation. The density medium has low viscosity and can be made isotonic for virtually any use. Gradients have proved useful in both human and experimental animal studies, and high percentage yields allow  for separations from small cell numbers. Methods are described for separation of whole blood and lymphoid subpopuctions. The cytoxic capability of various density fractions was evaluated for mixed lymphocyte culture-induced allogeneic killing and spontaneous, so-called "natural" killer cell activity. The lower density associated with blast transformation allows for significant enrichments of stimulated cells from in vitro cultures. Higher thymidine incorporation, restimualtion in mixed lymphocyte reactions, and greater cytotoxic capacity are associated with these "blast" fractions.


The interaction of cadmium-binding proteins (Cd-bp) and progesterone in cadmium-induced tissue and embryo toxicity.

Previous work indicates that a dimer of Cd-thionein (Cd-bp-D, 19,000 MW) is involved in the hereditary resistance to Cd-embryotoxicity seen in the inbred NAW/Pr (NAW) mouse strain. Cd-bp-D is not detected in virgin females after Cd exposure and is detected only after the first 24 hours of exposure to Cd in an inbred strain (C57BL/10ChPr) susceptible to Cd-induced embryotoxicity (Wolkowski, '74; Wolkowski-Tyl, '78). Since progesterone (P) is critical for maintenance of pregnancy in mice, we have studied the possible relationship between this hormone and Cd-bp-D production. As a model system, was examined effects of Cd treatment on Cd-bp synthesis in NAW males. It was anticipated that this model could provide information bearing not only on the relationship between P and Cd-bp-D production, but also on that between Cd-bp-D and Cd toxicity, since a single sc injection of CdCl2 causes typical testicular hemorrhagic necrosis in NAW males, and these animals make only metallothionein and not Cd-bp-D. NAW males were, therefore, given P (0.1 g/Kg bw) and then exposed to Cd. Sephadex gel chromatography (G-200) of liver cytosol from animals killed 24 hours later detected only Cd-bp-D. Testes of these males did not show hemorrhagic necrosis. Since the adrenals of male mammals release P in response to stress, NAW males were stressed by repeated sesame oil or propylene glycol injections (5 ml/Kg bw), or the adrenal was stimulated directly with injections of ACTH (100 IU/Kg bw) for seven days prior to Cd exposure. All methods tested which significantly elevated serum P levels (as confirmed by radioimmunoassay), also resulted in production of Cd-bp-D and absence of testicular hemorrhage in Cd-treated NAW males. Suppression of P release by injection of dexamethazone or corticosterone or by adrenalectomy resulted in testicular hemorrhage and production of only metalicthionein after Cd exposure. The relevance of the interaction between P, Cd-bp-D and protection against Cd-induced toxicity seen in the model system was supported by analysis of serum P levels in pregnant females; elevated levels were seen in resistant (NAW dams on day 10 of gestation and significantly lower levels seen in dams from a Cd-sensitive strain.


Small cell carcinoma: combined approach to treatment.

Fifty-six patients with untreated small cell carcinoma of the bronchus were treated with three courses of chemotherapy (cyclophosphamide, vincristine, and procarbazine and methotrexate) and assessed for response. Thirty-one patients (55.4%) were classified as responders; they were given a course of radiotherapy and were then randomly allocated to continued cyclical chemotherapy or not further chemotherapy until relapse. Non-responders to chemotherapy were treated with radiotherapy or palliatively. The median survival was 10.5 months in responders and 6 months in non-responders (P less than 0.01). The one-year survival in responders was 42%. There was no statistical difference in survival between patients treated with continued chemotherapy and those treated at relapse. Sixty-nine per cent of patients experienced no side effects from chemotherapy. Three indicators of non-response to chemotherapy were identified--exercise tolerance at diagnosis, macroscopic liver metastases, and inappropriate ADH secretion.


Platelet function and serum high density lipoproteins.

Two groups of healthy male subjects one with low (mean 1.03 mmol/l), and the other with normal (mean 1.80 mmol/l), concentrations of high density lipoprotein (HDL) cholesterol in plasma were investigated. A series of in vitro platelet function tests were performed in order to investigate a possible connection between low HDL concentration, which has been defined as an independent risk factor for the occurrence of CHD, and disturbances in platelet function. No significant differences in platelet function were observed between the two groups. A positive correlation was found between platelet factor 3 activity both in platelet rich and platelet poor plasma and the plasma concentration of HDL.


Macrophage inhibition of collagen-induced platelet aggregation.

Collagen was incubated with cells or media fractions of mouse peritoneal macrophage cultures, and its aggregating effect on human platelets was tested. Incubation with lysates of cultured cells completely abolished the normal collagen-induced platelet aggregation, while incubation with media fractions only caused partial inhibition. The latter inhibition was more pronounced after macrophage phagocytosis of latex particles, while endocytosis of endotoxin had no effect. Corresponding macrophage cultures were also tested for specific collagenase activity, using 14C-glycine labelled collagen as substrate. Collagenase activity was found in the culture media fractions only, and the enzyme activity could be enhanced by endocytosis of latex as well as endotoxin. It appears that the effect of macrophage lysates and media on collagen-platelet interaction cannot be ascribed only to secretion of collagenase from macrophages.


Effects of dithiocarb and (+)-cyanidanol-3 on the hepatotoxicity and metabolism of vinylidene chloride in rats.

The acute hepatotoxic effects of vinylidene chloride (VDC) were evidenced by measurement of the increase in the serum levels of the aminotransferase (GPT) and sorbitol dehydrogenase (SDH), hepatic glutathione (GSH) depletion and histological examinations in rats. The hepatoprotective agents dithiocarb and (+)-cyanidanol-3 proved well able to antagonize these toxic effects of VDC. While dithiocarb inhibited the in vivo metabolism of VDC in a closed exposure system, (+)-cyanidanol-3 had no influence at all. These findings substantiate the role of the microsomal monooxygenase system in the metabolism and hepatotoxicity of VDC. The mechanisms by which dithiocarb and (+)-cyanidanol-3 act as antihepatotoxic agents are different: the inhibition of the metabolic activation by dithiocarb and free radical-scavenging by (+)-cyanidanol-3.


Quantitative serum lipoprotein electrophoresis in Nigerians.

In response to the growing demand for lipid profiles in Nigerian patients with suspected ischaemic heart disease, cerebro-vascular accident and hypertension, serum lipoprotein electrophoresis based on ozone-Schiff staining technique and densitometric scanning has been developed. Out of 93 serum samples 80 showed three distinct bands in the electrophoretic patterns; nine showed two distinct bands and four had unusual migration characteristics. Mean percentages of the three bands in the common pattern were 43.5, 22.8, and 33.7 for alpha pre-beta and beta lipoproteins respectively. The relative preponderance of alpha lipoproteins in Africans contrasts with the findings in Europeans. It supports the view that high serum alpha lipoproteins may be protective against ischaemic heart disease in contrast to the atherogenic role of beta lipoproteins.


DNA and oestradiol receptor distribution in human breast cancer.

The content of DNA and oestradiol receptors was measured in two different samples taken from the same tumoral mass of 33 primary infiltrating ductal carcinomas. The results were related to the total cytoplasmic protein and to the DNA of the tissue. The results showed that variations of both DNA and oestradiol receptors are frequent and often significant. The reliability of analyses carried out on a single biopsy is discussed.


Prolactin receptor in human mammary carcinoma.

The specific binding of labelled human prolactin was determined in 83 human breast carcinomas. Twenty-seven tumors (32.5%) contained specific binding for prolactin of at least 1% and were considered prolactin receptor positive. The binding was found linearly related to membrane protein concentration and specific only for lactogenic hormones. By Scatchard analysis the dissociation constant appeared similar to that observed in other target tissues, with a low binding capacity.


A field trial with a multicomponent inactivated respiratory viral vaccine.

An inactivated virus vaccine containing strains of parainfluenza type 3 (PI3), bovine adenovirus type 3, reovirus type 1, bovine virus diarrhoea (BVD) and infectious bovine rhinotracheitis (IBR) viruses was tested in a group of 58 calves reared in a semi-intensive management system. Following vaccination, 1/30, 14/30 and 17/30, showed significant rises in antibody titre to reovirus type 1, adenovirus type 3 and IBR respectively. None of the animals showed significant serological response to PI 3 and BVD. In the control group, 2/28, 1/28, 6/28 and 3/28 developed antibody responses to reovirus type 1, BVD, adenovirus type 3 and IBR respectively. Microbiological examination revealed the presence of a wide variety of commensal bacteria and Mycoplasma bovirhinis in both groups. Analysis of the records of clinical examinations indicated that the respiratory tract infections occurred among the calves at between 50 and 80 days after arrival at the farm, and that there was no significant difference between the test and the control groups. A number of animals had maternal antibodies to the various components of the vaccine present before the trial commenced and these antibodies appeared to interfere with the subsequent serological response to the antigen challenge. The vaccination schedule recommended by the manufacturer does not entirely circumvent this problem.


[Influence of glutaraldehyde on Na+, K+-ATPase activity of brain tissue and enzymic preparations under conditions of electron cytochemical analysis].

The glutaraldehyde inhibitory influence on Mg2+, Na+, K+-ATPase activity of the brain tissue is shown to increase with a rise in the glutaraldehyde concentration; a direct dependence is established between the degree of inhibition and the treatment time and temperature. The glutaraldehyde inhibitory effect on the activity of the brain microsomal fraction Na+, K+-ATPase preparations under the same treatment conditions is considerably higher. A histochemical detection of the cerebral cortex Mg2+, Na+, K+-ATPase was carried out under optimal conditions of fixation, and a conclusion is made on possibility of using glutaraldehyde as a fixator in histochemical investigations.


[Serological diagnosis of IBR. Comparative studies of cattle sera in the virus neurtalization and passive hemagglutination reactions].

Three hundred eighty six cattle serums were simultaneously investigated by virus neutralizing (VN) and passive hemagglutination (PSH) reactions. Results obtained by the two methods coincided in 94% of the sermums studied and all serums with VN antibodies had also antibodies which can be established by the PSH reaction. About 6% of the positive reagents for PSH reaction were negative for the VN reaction. These were serums containing early antibodies (IgM) or low-titer antibodies of the IgG class, and were subtreshold for the VN reaction. The antibody titers determined by the PSH reaction were 7 to 10 times higher than these determined by the VN reaction, while their mean titer in serums with minimum VN index 1.0 was 1:23. The conclusion is drawn that PSH can be used as an independent serological test.


[Experimental selection of weakly virulent strains of Aujeszky's disease virus].

An attempt was made to select slightly virulent strains by consecutive passages in cell cultures and use of extreme dilutions applying the temperature test (heating up to 50 degrees C for 1 hour). Initial and final fractions were compared for lethality and pruritus on rabbits and studied by the plate method. Levelling of the cytopathogenic titer after heating heated and unheated virus was achieved in laboratory samples isolated from lungs, and in two strains -- isolated from brains. This was achieved in the various strains following a varying number of passages, less in those isolated from lungs and more in those isolated from brains. Initial strains isolated from brains cause, while the lines produced by them do not cause, scratching in the infected rabbits. Temperature sensitive strains (identified by this test as virulent) produce mainly large (1.2 mm) plates, while the vaccine strain and the strains identified by the test as slightly virulent produce mainly small (0,8 mm) plates.


[Isolation of the bovine herpesvirus-3 from sick calves].

Three herpes strains identified as herpes virus-3 were isolated in revolving test-tubes with cell cultures of nasal samples and lung parenchyma from diseased calves at the time of a respiratory enzooty. Large crystalloid aggregates of herpes virus virions were observed in the nucleus and in the cytoplasma by the electron-microscopic investigation performed on infected cell cultures. The experimental infection of calves with the isolated strain produced slight symptoms of respiratory disease in single calves only. The infected calves were susceptible to a following infection with cattle herpes virus-1.


[Serological diagnosis of IBR. Passive hemagglutination microreaction].

The preparation and standardization of the ingredients for PHA is described in detail. Formalin treated erythrocites in a 10% concentration treated with 1:20 000 tannin were used. The antigen concentrated by ultracentrifugation properly sensibilizes the erythrocites and their titer does not change in case of its dilution up to 1:8. The non-concentrated virus antigen has a low covering potential only if it is either not diluted, or in a 1:2 dilution and sensibilizes the erythrocites to a lesser extent. The great advantage of the use of concentrated antigen is that it equalizes antigens obtained from various lots. The optimal time for loading by erythrocites is 1 h, and a longer sensibilization has no influence on the effect of loading. It was proven that the sensibilization of formalin treated erythrocites allows long storage of the produced antigen. The tannin treated formalinized erythrocites can be stored up to 20 days without any change in their ability to be antigen covered, while the sensibilized erythrocites are not changed for 6 days and up to the 30th day their titer is twice lower, but this is of no practical importance.


N-mineralization of formaldehyde-releasing N-compounds.

The mineralization of several N-compounds, such as MMU, DMU, and HMT, as well as their respective nitrificide action, have been investigated. It has been proved that the amount of available formaldehyde, as well as its rate of release, are responsible for the specific action of these compounds on the N-mineralization process. Factors affecting the rate of release, namely molecular structure, solubility, and temperature, are discussed as well. Importance of these N-compounds as slow-release N-fertilizers based on a new molecular-microbiological approach is described.


Enhancement of the antiviral activity of pyrimidine derivatives against mengovirus by visible light.

Eleven pyrimido-pyrimidine derivatives, seven with significant antiviral activity against Mengovirus, five against Coxsackie B1 virus and four antiviral negative compounds were tested for their photosensitizing ability. All seven compounds with antiviral activity in vitro showed an enhanced antiviral action against Mengovirus under irradiation with visible light, a fact that may be caused by photodynamic processes. It was tried to correlate the oxidation potentials of sensitizers with their photodynamic activity. By means of mass-spectrometric investigations, molecular fragmentation was examined following thin layer chromatography (TCL) before and after irradiation. Furthermore, binding affinity to biopolymers (BSA and RNA) was investigated to reveal conformity in differences of antiviral activity. The main results are the following: 1. Generally, strong antiviral activity can be correlated with strong binding affinity. 2. No significant correlation could be detected between oxidation potentials of antiviral compounds and their enhanced antiviral activity under irradiation conditions, although in some cases sensitizer with higher oxidation potentials are more effective than those with lower ones. 3. The lower the photostability of the compounds the higher was the light-induced antiviral activity. 4. No alteration of the molecular ion peak and fragmentation pattern before and after irradiation was indicated by means of mass-spectrometry and TLC using fairly comparable conditions.


[Nonspecific prophylaxis and therapy of Pseudomonas aeruginosa wound-infections with paramunization using a mouse-model (author's transl)].

The effectiveness of paramunization as a antigen nonspecific method to activate mechanisms against wound-infections due to Pseudomonas aeruginosa was studied using a model of direct infection of mice with a "mice-pathogenic" Ps. aeruginosa strain on artificially set wounds. Active paramunization by means of a biological inducer "PIND-AVI" (M-HP 438) significantly reduced the mortality rate between treated and placebo animals. The best results were obtained by parenteral prophylactic application. A four times repeated injection of PIND-AVI before the wound-infection reduced the mortality rate from 80% (placebo animals) to 26.6%. Almost equally good results were obtained by clinically useful therapeutic application of the preparation. A four times repeated treatment of the mice after wound infection lead to a decrease of mortality rates from 86.6% to 36.6%. The paramunization inducer PIND-AVI caused no side effect in any of the experiments. The mode of inducer action in Pseudomonas aeruginosa wound infections appears to be complex. Increased phagocytosis by nonspecific opsonisation, increased macrophage activity and concurrent stimulation of the lymphopoetic system could possible occur. On the other hand the nonspecific action of mediators could also play a role due to the inducer stimulated T-cells and cellular antigens of Pseudomonas aeruginosa. However both mechanisms in cooperation with specific and nonspecific humoral factors are probably interacting together. To what extent a simultaneous synthesis resp. release of endogeneous interferon plays a role is not known.


The effect of nalidixic acid on microaerophilic and anaerobic bacteria. Special study of clostridia.

Study of the sensitivity of 141 strains of anaerobic and microaerophilic bacteria to nalidixic acid shows that few bacteria are inhibited by low concentrations (Veillonella, Eikenella, most of the Clostridia). Nalidixic acid appears to be bactericidal with respect to Clostridium perfringens, and its point of attack in the DNA is probably different from that of metronidazole.


[Purification of horseradish peroxidase conjugated IGG by affinity chromatography. Comparison of glutaraldehyde and periodate conjugation].

By affinity chromatography of the coupling mixture on Concanavalin-A-Sepharose unlabeled IgG is completely removed. HRP conjugated IgG is then separated from free HRP by gelfiltration. Glutaraldehyde conjugation yielded 20 to 68% unlabeled IgG, depending on the duration of glutaraldehyde addition, and periodate conjugation yielded 10 to 28% unlabeled IgG. By this method fractions of conjugates with 3fold spec. act. were obtained by the glutaraldehydemethod and with 1.6fold spec. act. by periodate method as compared with gelfiltration. The periodate method guarantees a 3fold higher yield of HRP-labeled IgG compares with glutaraldehyde method. Both methods produce conjugates of the same spec. act. although the denaturation of HRP is much greater at periodate conjugation. When applying of affinity chromatography and gelfiltration. Crude HRP with 7% spec. act. of purified HRP resulted in conjugates having a spec. act. of 85% of those with purified HRP.


Lack of sequence homology between the nucleic acids of Rauscher leukaemia virus and polyoma virus Y8e produced simultaneously in a continuous mouse cell line.

In a continuous cell line (Y8e) from spleen and thymus cells of mice, infected with RLV, the presence of both RLV and polyoma virus Y8e in a single cell could be demonstrated by electron microscopy. A comparison of the nucleic acids of RLV and polyoma virus from Y8e cells by two molecular hybridization methods showed lack of sequence homology between the viral nucleic acids.


Ontogenetic development of 3,5-adenosine monophosphate phosphodiesterase in guinea pig and rat ileum.

The postnatal development of phosphodiesterases with low and high Michaelis constants in guinea pig and rat ileum was studied. The tow types of phosphodiesterases were found to increase their activity post partum, and this increase was particulary pronounced in the rat. The rise in the enzyme activity took place mainly at the expense of the increase of the phosphodiesterase with high Km. In addition to the quantitative differences in the phosphodiesterase activity of yound and adult animals, considerable age differences were also observed in the sensitivity of the enzyme to inhibitors and activators. These data can contribute to the explanation of the differences in the action of some drugs influencing the phosphodiesterase in young and adult organisms.


Combined test of hypothalamic-pituitary function in growth-retarded children treated with growth hormone. II. Secretion of LH, FSH, TSH, prolactin and ACTH.

A total number of 23 patients treated with human growth hormone were retested by use of a combined pituitary stimulation test. Plasma concentrations of GH, FSH, LH, TSH, T4, T3, prolactin (PRL), ACTH and cortisol were measured before and after stimulation with hypoglycemia, TRH and LHRH. The test was performed in patients with persistent GH deficiency (group A) and patients with transitory GH deficiency (group B). In group A a normal pubertal development was found in three patients, whereas in prepubertal subjects the FSH/LH responses were smaller than those of prepubertal patients in group B. Also plasma ACTH increase was less pronounced in group A patients than in group B. In contrast, the plasma TSH and PRL responses were more sustained in group A than in group B. The secretory pattern of TSH and PRL was comparable in the two groups of patients. Thus, in patients with persistent GH deficiency additional multiple disturbances of the hypothalamic-pituitary function often appeared whereas in most patients with transitory GH deficiency the combined pituitary test was normal at the reinvestigation.


Low urinary estriol during pregnancy caused by isolated fetal ACTH-deficiency.

In a 34-year-old pregnant woman, serum HPL and urinary HCG were normal, but urinary estriol was repeatedly low. A normal boy was delivered after 38 week gestation. During the neonatal period, he had hypoglycemia, muscular hypotonia and transient hyperbilirubinemia. The ACTH-test was normal, but the THS-response to metyrapone was low. Serum ACTH did not respond to insulin and metyrapone. Growth hormone, TSH and gonadotropin responses to stimuli were normal. Treatment with hydrocortisone resulted in disappearance of the symptoms. It is concluded that fetal ACTH-deficiency is one of the specific endocrine causes of low maternal estriol.


Growth in Turner's syndrome: spontaneous and fluoxymesterone stimulated.

Spontaneous growth was analysed in a group of 55 girls with Turner's syndrome and various karyotypes. Their variation in height and its dependence on parental height were similar to that of normal girls. At all ages, the 45,X karyotype was associated with slightly greater mean stature than the other karyotypes together. The bone ages lagged progressively behind from 10 years onwards. Twenty-five patients aged between 9.1 and 17.2 years were given fluoxymesterone, 0.06-0.17 mg/kg daily, for at least 1 year. Their height velocities increased significantly. This brought about a clear psychological benefit. Their final heights were predicted before and after therapy, with a new method based on the spontaneous growth and bone maturation of our patients. The response was individually variable but, on average, the patients gained in predicted height from the therapy. This effect was not lost during a posttreatment year. Abnormal lowering of the voice occurred in patients receiving greater than or equal to 0.15 mg/kg fluoxymesterone daily, but never with less than 0.13 mg/kg. No other adverse effects appeared. Thus, fluoxymesterone is useful for promoting growth in girls with Turner's syndrome.


Acute response of parathyroid hormone in congenital osteopetrosis.

Indices of calcium and phosphorus metabolism were studied in 3 children with osteopetrosis before and after infusion of bovine parathyroid hormone extract. Basal plasma concentrations of calcium, alkaline phosphatase and 25-hydroxy vitamin D tended to be low. Plasma immunoreactive PTH levels were at the upper normal range in two patients. A marked increase in urinary cyclic AMP in all patients was solely due to an increase in the nephrogenous cAMP. After vitamin D treatment urinary cAMP was essentially unchanged with the same preponderance of nephrogenous cAMP. Following PTH infusion plasma cAMP showed a brisk rise. There was also a prompt rise in urinary cAMP and a distinct decrease in the calcium to sodium clearance ratio indicating increased calcium reabsorption. Phosphaturic effect was only observed when PTH was given in the highest dose level. The findings are consistent with a state of low grade hyperparathyroidism which could not be related to the plasma levels of 25-hydroxy vitamin D or calcium.


The effect of sympathicomimetic agents on carbohydrate metabolism of planaria.

Epinephrine, ephedrine, dopamine and isoproterenol considerably influence carbohydrate utilization in planaria. Dopamine alone and in combination is the most potent, while ephedrine the least effective in this respect. The finding of increased responses to combined treatment supports the view of the existence of different receptor sites for the sympathicomimetic drugs. The results substantiate conclusions concerning the phylogenesis of hormone receptors.


Sensitivity of European microtines to injected CdCl2.

Basic histologic and metabolic indices of damage to the testes by CdCl2 were observed in six species of rodents: Lagurus lagurus, Pitymys subterraneus, Microtus oeconomus, M. agrestis, Clethrionomys glareolus (and Mus musculus for comparison). All species showed discoloration of the subcapsular tissues and damage to the testicular parenchyma. Testes of all species receiving CdCl2 utilized from 43% to 76% less oxygen than did those from untreated individuals. Cadmium treatment preceded few significant temperature shifts. The voles observed experienced differential sensitivity to CdCl2 and should be quite suitable for further studies of the mechanism of action of CdCl2 as a testicular poison.


Inhibition of the stimulatory effect of ACTH on adrenal and ovarian blood flow by indomethacin in the dog.

In anoestrous dogs heated thermocouples were inserted into one of the adrenals and ovaries. The basal adrenal and ovarian blood flow was recorded and enhanced by 1.2, 2.5 or 5.0 micrograms/kg of intravenous ACTH before and after the intravenous administration of 20 or 50 mg/kg of indomethacin. Indomethacin, depending on the dose and on the time elapsing after its administration inhibited the enhancing effect of ACTH on adrenal and ovarian blood flow. Since indomethacin is a specific inhibitor of prostaglandin synthesis it has been suggested that prostaglandins act as mediators in the vasodilatation elicited by ACTH in the canine adrenal and ovary.


Involvement of dopaminergic receptors in action of corticotropin releasing factor (CRF) under in vitro conditions.

ACTH release under the effect of median eminence extract (ME) was studied in both incubation and superfusion experiments. ACTH content of the incubation medium was measured by radioimmunoassay or by the corticosterone production of trypsinisolated adrenocortical cells. Dopamine at low concentration led to a slight increase of basal ACTH secretion, while at higher concentration failed to influence ACTH release. The dopamine agonist CB-154 produced a significant rise of ACTH secretion and augmented the ME extract-induced increase of pituitary ACTH release. Chlorpromazine and haloperidol suppressed basal ACTH secretion and inhibited the ME extract-induced release. The simultaneous administration of CB-154 and haloperidol into the incubation medium prevented the haloperidol-induced inhibition of ACTH release. The observations indicate that dopaminergic receptors play a role in the activation of CRF-induced ACTH secretion under in vitro experimental conditions.


Circadian peculiarities of daytime sleep in 1 to 6 months old infants (electrophysiological study).

A longitudinal study of 6 normal, full-term infants was carried out. Polygrams of three subsequent daytime sleep periods (morning, noon, afternoon) were recorded at monthly intervals. It was established that: (a) Over the first 3 postnatal months the noon period was the longest (98.96 min), and each sleep period consisted most frequently of 2 cycles. Over the second 3 months the afternoon period shortened significantly from 68.69 min to 41.92 min and consisted exclusively of 1 cycle. (b) The delta sleep percentage was the greatest (52%) over the first 3 months in the morning period and over the second 3 months it was almost equal in all the three periods. The paradoxical sleep percentage over the first 3 postnatal months was equal in all the three periods, but over the second 3 months it became smaller in the afternoon period, (c) The heart rate values were lowest in the morning period, this phenomenon being clearly expressed during the delta sleep. One minute time (in sec), occupied by REMs, was evaluated and it was found that this parameter was not influenced by age and timing of sleep periods, but there existed positive correlation between this time and the duration of the paradoxical phase under estimation.


Inhibitory actions from low and high threshold cutaneous afferents on groups II and III muscle afferent pathways in the spinal cat.

The inhibitory effects caused by volleys in cutaneous afferents on the transmission through some polysynaptic segmental pathways activated by high threshold muscle afferents were studied in chloralose anesthetized, spinal cats. Pathways studied were groups II and III to motoneurones as well as group II to primary afferents. The results suggested that two different mechanisms were involved. One mechanism, with a very slow time course (duration more than 400 ms), is suggested to be an example of presynaptic inhibition between different primary afferent systems. This mechanism required high threshold (greater than or equal to 1.6T) conditioning shocks, and appeared simultaneously with the component II dorsal root potential being evoked by the cutaneous afferent volley. The other mechanism, with a faster time course (duration always below 300 ms), was dependent upon low threshold (less than or equal to 1.5T) cutaneous conditioning volleys. This inhibitory interaction could not be ascribed to the same presynaptic mechanism, but is suggested to be an example of postsynaptic inhibition at an interneuronal level. The presumed disynaptic excitatory pathway from group II muscle afferents to flexor motoneurones was not inhibited by cutaneous conditioning shocks, but could on the contrary be facilitated by activity in low threshold cutaneous afferents, probably at the only interneurone involved in this group II pathway.


Adenosine 3',5' cyclic monophosphate, calcium and magnesium excretion in ethanol intoxication and hangover.

Effect of ethanol on adenosine 3', 5' cyclic monophosphate (cAMP), calcium (Ca) and magnesium (Mg) excretion was studied in controlled clinical conditions in man. Seven male volunteers served as their own controls. In 5 subjects cAMP excretion was primarily suppressed by ethanol. Ethanol appeared to have a biphasic effect on Ca excretion, an initial stimulation followed by a conservation phase. Mg excretion was stimulated by ethanol in 5 subjects. Subjects having nausea and vomitus and the most severe hangover symptoms had the lowest urinary Ca excretion and the lowest imitial cAMP excretion. Ca and Mg metabolism and the susceptibility of the body to the toxic effects of ethanol appeared to be interrelated.


Modification of human pain threshold by specific tactile receptors.

The effects of conditioning vibrotactile stimulation of particular tactile receptor groups on thresholds to painful electric stimuli were studied in seven healthy adults. Preferentially Pacinian afferents were activated with conditioning sinusoidal vibration of 240 Hz at 20 and 200 micrometers amplitudes and preferentially non-Pacinian tactile fibers were activated with conditioning sinusoidal vibration of 20 Hz at 200 and 400 micrometers amplitudes. None of the subjects showed pain threshold elevation during activation of non-Pacinian tactile fibers. However, 6 of the 7 subjects showed significant pain threshold elevation with conditioning vibration stimulus of 240 Hz at 200 micrometers amplitude, and 4 subjects showed significantly elevated pain thresholds with conditioning stimulus of 240 Hz at 20 micrometers amplitude. It is concluded that the activation of Pacinian afferents causes inhibition of pain conducting pathways.


Efflux of cyclic AMP, prostaglandin E2 and F2 alpha and thromboxane B2 in leg lymph of rabbits after scalding injury.

Leg lymph was collected from pentobarbital anaesthetized rabbits before and after scalding injury of the paw (75 degrees C for 20 s), and the contents of cyclic AMP (cAMP), prostaglandin E2 (PGE2) and PGF2 alpha and thromboxane B2 (TXB2) in lymph were determined. After injury lymph flow increased about four times. The maximal rate of flow was found between 30 and 60 min after scalding. The efflux of cAMP and immunoreactive iPGE2, iPGF2 alpha and iTXB2 also increased. The maximum values were detected at approximately 0-30, 30-60, 30-60 and 180-240 min, respectively, after the injury. The output of cAMP, iPGE2 and iPGF2 alpha and iTXB2 in lymph of the contralateral non-scalded paw remained low throughout the experiments. When rabbits were injected with indomethacin (2.5 mg/kg) or diclofenac sodium (2.5 mg/kg) immediately after the scalding injury, the efflux of cAMP, iPGE2 and iPGF2 alpha were low. Lymph flow was markedly reduced after treatment with diclofenac sodium; treatment with indomethacin did not significantly affect lymph flow. The results suggest a prostaglandin-dependent formation of cAMP following scalding injury which may be related to the initial responses to scalding.


Haemagglutination-inhibiting antibodies to B.K. virus in Hungary.

Antibody inhibiting the haemagglutination by B.K. virus was found in 64% of the serum samples collected from 949 subjects in Hungary. The frequency of seropositivity was the lowest (17%) among infants 1 and 2 years of age and the highest (93%) in the age group 16 to 20 years. The subsequent age groups showed a slow continuous decline. The age-distribution curve drawn on the basis of the geometric means of titres has two peaks, one between 3 and 5 years of age and another between 21 and 30 years of age, reaching 6.74 and 7.05, respectively, as expressed in terms of log2 units. The prevalence of haemagglutination-inhibiting antibody to B.K. virus in Hungary and its distribution by age are similar to those reported from other European countries.


Isolation of cytopathogenic Rotavirus from neonatal calves.

Rotavirus was isolated in rolled calf kidney cultures from the intestinal contents of a calf suffering from diarrhoea. The cytopathic effect was demonstrated in native cultures as well as in stained preparations. The affected cells were elongated, became sickle-like in shape, disintegrated and detached from the monolayer. Virus-specific direct immunofluorescence ran parallel, in both time and intensity, with the cytopathic effects. Immuno-electron microscopy showed aggregated virus particles corresponding in size, shape and structure to Rotavirus virion.


Effect of acupuncture on adrenocortical hormone production: I. Variation in the ability for adrenocortical hormone production in relation to the duration of acupuncture stimulation.

The effect of acupuncture on adrenocortical hormone production in adult female albino rabbits of New Zealand strain was investigated. There was significant increase compared to the control cortisol and corticosterone following application of acupuncture at the locus Tsu-San-Li. In the non-locus group, such treatment resulted in only a transient raise followed by a significant fall in cortisol and corticosterone. Acupuncture treatment with electric current for 1 hour proved to be unique in that it gave rise to no biphasic change in the 14C uptake of the steroids. These results indicate that adrenal production of corticosterone and cortisol was enhanced by acupuncture and electroacupuncture at the Tsu-San-Li locus.


Focal nodular hyperplasia and adenoma of the liver. A pediatric experience.

Focal nodular hyperplasia and adenoma of the liver together represent approximately 2% of all primary hepatic tumors and tumor-like lesions in childhood. This study reports the clinical and pathologic features of focal nodular hyperplasia in three children, all females between 27 months and 15 years of age with asymptomatic hepatomegaly. In contrast, massive hemoperitoneum from a ruptured, hemorrhagic mass was the presentation of the hepatic adenoma in a 14-year-old girl. There was no history of administration of steroids in these four children. Angiography in two cases (one case each of focal nodular hyperplasia and hepatic adenoma) revealed hypervascular lesions with abnormal tortuous vessels suggesting a malignant tumor. Pathologically, the adenoma and focal nodular hyperplasia were readily distinguishable and the necessity for this differentiation was reviewed. All four patients are currently doing well.


Cystic nephroma (so-called polycystic Wilms' tumor) of childhood: a CT-pathologic study.

The unique CT features, heretofore unreported, of a case of cystic nephroma of childhood are presented. The concept that cystic nephroma is part of thc diagnosis of cystic nephroma can be made utilizing excretory urography and computerized tomography.


Histochemical analysis of Langerhans' cells.

Single cell suspensions of epidermal cells from guinea pigs were analyzed histochemically for the presence of the following enzymes: 5'-adenosine triphosphatase, nonspecific esterase, specific esterase, myeloperoxidase and leukocyte alkaline phosphatase. A population of cells was positive for nonspecific esterase, leukocyte alkaline phosphatase, and 5'-adenosine triphosphatase. This population was identified as Langerhans' cells because the number of cells stained for these enzymes paralleled the number of Langerhans' cells in the suspension. These same enzymes were shown to be present in guinea pig leukocytes of the mononuclear-phagocytic series, suggesting that they and Langerhans' cells may have a precursor in common.


Association of islet cell carcinoma of the pancreas with subcutaneous fat necrosis.

A 57-year-old man with generalized, subcutaneous fat necrosis was found at autopsy to have islet cell carcinoma of the pancreas. The histologic diagnosis of islet cell carcinoma was confirmed by the finding by electron microscopy of characteristic intracytoplasmic granules within pancreatic neoplastic cells. Lipase levels were elevated in serum as well as in tissues in areas of subcutaneous fat necrosis. Enzyme histochemical stains for alkaline and acid phosphatase, leucine, aminopeptidase, succinic dehydrogenase, and indoxyl and nonspecific esterase were positive in the areas of subcutaneous fat necrosis. This is the first report of the association of islet cell carcinoma of the pancreas with generalized subcutaneous fat necrosis.


Brain's entropy partitions.

Since the brain processings can be expressed by ballistic or ramp systems, the brain entropy is herein analysed in respect to the distributions of the internal ((MS)1) and external messages ((ME)E) conveyed through the sensory systems. In such a context, it is shown that a first partition (through a primary folding) is established between the neural systems involved in the processings related with the external (RSE) and internal (BSI) environments. Indeed, secondary foldings in both RSE and BSI due to non-homogeneities in (MS)E and (MS)1 are discussed. Finally, the results are extended to the neural phenomena during the brain's embriogenic growth.


Kinetic studies of the blood serum xanthine dehydrogenase inhibition by alloxan (2,4,5,6-tetraoxypyrimidine 5,6-dioxyuracil).

Alloxan is an inhibitor of the enzyme xanthine: NAD+ oxido reductase (E.C.1.2.1.37). Alloxan acts as an electron acceptor and competes "in vitro" with the tetrazolium salt used as electron acceptor in the assay system used for the determination of the dehydrogenase activity of the enzyme. The alloxan inhibition was reverted when phenazine methosulfate (PMS) was added to the system.


Neurogenic tumors of the head and neck in children.

A retrospective analysis of deep-seated neurogenic tumors of the head and neck in children seen at the University of Michigan Medical Center was done.  There were 22 patients--10 with plexiform neurofibromas, 3 with neurofibromas, 2 with esthesioneuroblastomas, 2 with neurilemmomas, 1 with neurogenous sarcoma, and 4 with neuroblastomas who presented with a mass in the head and neck.  Delay in diagnosis and treatment was common, and merely allowed the tumor to enlarge.  We feel that surgical extirpation is the preferred treatment for benign or malignant neurogenous tumors, with adjuvant therapy in other forms for malignant tumors only.


Prophylactic subcutaneous total glandectomy for mammary cystic disease, with immediate primary breast reconstruction.

Seventeen whole-breast tissue specimens of mammary cystic disease were prepared using Marcum's whole-mounting technique. Histological studies were performed on from 10 to 19 slides of tissue containing the suspected pathological foci. Cysts in the cleared breast tissues were numerous, and cyst walls were covered by single-layered duct epithelial cells with some apocrine metaplasia. The most notable findings were frequent minute carcinomatous foci next to the scattering of mammary cysts. The incidence of carcinoma among 17 mammary cystic disease patients was 8, or 47%. Each patient had from one to six minute carcinomatous foci--less than 1 mm in diameter and noninfiltrating. All patients were treated by subcutaneous total mastectomy and immediate breast reconstruction with the Natural-Y mammary prosthesis. In all 17 the results were good. With no hazardous complications. Steps toward immediate breast reconstruction are as imperative psychologically as the total glandectomy is clinically.


Participation of lysine-sensitive aspartokinase in threonine production by S-2-aminoethyl cysteine-resistant mutants of Serratia marcescens.

S-2-Aminoethyl cysteine (AEC) reduced both growth rate and final growth level of Serratia marcescens Sr41. The growth inhibition was completely reversed by lysine. AEC inhibited the activity of lysine-sensitive aspartokinase to a lesser extent than lysine. The AEC addition to the medium lowered not only the level of lysine-sensite aspartokinase but also those of homoserine dehydrogenase and threonine deaminase, whereas lysine repressed the aspartokinase alone. To select mutations releasing lysine-sensitive aspartokinase from feedback controls, AEC-resistant colonies were isolated from strains HNr31 and HNr53, both of which were previously found to excrete threonine on the minimal plates but not on the plates containing excess lysine. Two of 280 resistant colonies excreted large amounts of threonine. Strains AECr174 and AECr301, derived from strains HNr31 and HNr53, respectively, lacked both feedback inhibition and repression of lysine-sensitive aspartokinase. These strains produced about 7 mg of threonine per ml in the medium containing glucose and urea.


Effect of sample transport systems on survival of bacteria in ground beef.

The effects of two transport systems and cryoprotective agents on the survival of bacteria in ground beef samples were evaluated. Survival of Clostridium perfringens in ground beef samples after simulated transport (72 h) was higher (about 99%) in Dry Ice than in Trans Temp shipping units (-3 degrees C). There were no significant differences between the two transport systems in survival of coliforms, Escherichia coli, Staphylococcus aureus, or aerobic bacteria. Mixing ground beef samples at a ratio of 1:1 (wt/vol) with 10, 20, or 30% buffered solutions of dimethyl sulfoxide or glycerol before freezing improved the survival of C. perfringens and coliforms in both transport systems. Recovery of E. coli was significantly higher with the addition of 10% dimethyl sulfoxide before Dry Ice transport. Addition of 10% dimethyl sulfoxide resulted in a 100% recovery of both S. aureus and aerobic bacteria from ground beef after simulated transport in Trans Temp shipping units. The use of cryoprotective agents can improve the survival of bacteria during transport of ground beef samples.


Influence of stimulation of myocardial alpha- as well as beta-adrenoceptors on the effect of digoxin in isolated electrically driven rabbit papillary muscles.

On isolated electrically driven rabbit papillary muscle the cardiac glycoside digoxin was infused at driving rates of 0.5, 1.0 and 2.0 Hz. Two effective concentrations of digoxin were determined: 1. that inducing the maximal inotropic effect (maximal inotropic concentration) and 2, that causing cardiac arrest (toxic concentration). The influence of the alpha-sympathomimetic drug phenylephrine and for comparison that of the beta-sympathomimetic drug isoprenaline on either concentration of digoxin was investigated. 1. Stimulation of alpha-adrenoceptors by phenylephrine at a rate of 0.5 Hz significantly decreased the maximal inotropic as well as the toxic concentration of digoxin by about 36%; this decrease was maximal under the influence of the EC25 of phenylephrine and could be blocked by phentolamine. Phenylephrine did not alter the maximal inotropic effect of digoxin. At a stimulus rate of 1.0 Hz the EC75 of phenylephrine still diminished significantly the effective concentrations of digoxin whereas under these conditions the EC25 was ineffective. At 2.0 Hz stimulation of myocardial alpha-adrenoceptors had no effect anymore on either the maximal inotropic or the toxic concentration of digoxin. 2. In contrast, stimulation of beta-adrenoceptors by isoprenaline at a driving rate of 2.0 Hz resulted in a pronounced decrease of maximal inotropic and toxic concentration of digoxin while the maximal positive inotropic effect exerted by digoxin was found to be not altered by isoprenaline. The decrease of the effective concentration of digoxin caused by isoprenaline was abolished by pindolol. At a driving rate of 1.0 Hz this effect was slightly attenuated but was completely absent at 0.5 Hz. 3. From these results it can be concluded that stimulation of either adrenoceptor, alpha- and beta-, increases the effectiveness of the cardiac glycoside digoxin, i.e. diminishes the maximal inotropic as well as its toxic concentration. While stimulation of alpha-adrenoceptors is effective only at low rates of stimulation that of beta-adrenoceptors is vice versa at higher ones, thus supporting the view of different mechanisms underlying stimulation of alpha- or beta-adrenoceptors.


In vitro transformation of primary Chinese hamster cells by minichromosomes from Simian Virus 40.

Purified SV 40 DNA and SV 40 minichromosomes, either isolated from SV 40 infected cells (NaMi) or reconstituted from viral DNA and the H1 depleted calf thymus histone fraction (ReMi), were tested for their ability to transform primary Chinese hamster lung (CHL) cells. Using the agar suspension technique as the transformation assay, an approximately 200 fold increase in transformation activity for NaMi in comparison with the SV 40 DNA and an approximately 50 fold increased activity in comparison with ReMi could be demonstrated. Uptake experiments suggest that NaMi are taken up quicker and more efficiently from the CHL cells than ReMi or purified DNA. The transforming activity is most significantly using the calcium technique and the infected cells in higher passage numbers. Transformed colonies recovered after infection of CHL cells with SV 40 DNA, NaMi or ReMi revealed SV 40 related T-antigen in 90-100% of the cells tested. The number of T-antigen positive cells was stable over a period of 80 passages. This observation suggest a stable state of transformation by SV 40 DNA and SV 40 minichromosomes.


[Effect of bilateral and unilateral electroconvulsive therapy (ECT) on the composition of the cerebrospinal fluid (CSF). A possibility to calculate the intracellular redox changes of the brain in humans (author's transl)].

The changes in the composition of CSF and venous blood under the effect of bilateral and unilateral ECT were studied. The intracellular redox changes during the therapy were calculated on the basis of experimental data. Of the patients, 27 were treated with bilateral, another 27 with unilateral ECT. Just before the electroshock, the patients were paralyzed with intravenous succinylcholine chloride and were artificially ventilated with O2. The characteristic change we found was the augmentation of pyruvate concentration in CSF. This change and the calculated shift of NADH/NAD+ ratio in the direction of oxidation refer to the disturbance of the function of enzymes which play a role in the oxidation-reduction processes.


Comparison of hemagglutination-inhibition, complement-fixation and enzyme-linked immunosorbent assay for quantitation of human rotavirus antibodies.

The hemagglutination-inhibition test for detecting rotavirus antibody was evaluated by using simian rotavirus SA-11 as hemagglutinating antigen. Results show that the test is as sensitive as either complement fixation or the enzyme-linked immunosorbent assay for detecting antibody to rotavirus in human sera.


Oncogenic transformation of rat lung epitheloid cells by SV 40 DNA and restriction enzyme fragments.

Rat epitheloid lung cells were transformed with various preparations of SV40 dna using the Ca2+-precipitation technique. The amount of SV40 genetic information integrated into transformed clones was evaluated by DNA-DNA renaturation kinetics. The growth properties on plastic and in soft-agar were examined, as well as the ability to induce tumors in syngeneic new-born animals or in adult nude mice. One particular transformed line, which had received the Hpa II/BamH I A (59 per cent) fragment, was found to contain about 3 integrated copies of this fragment per cell and no significant amount of the Hpa II/BamH I B (41 per cent) fragment. This line which grew to high saturation densities and efficiently formed clones in low serum on plastic, produced tumors in both syngeneic rats and nude mice. Thus the Hpa II/BamH I A fragment, which mainly includes early viral information, was sufficient to impart these properties to rat epitheloid lung cells.


Nonarbo-togaviridae: comparative hydrodynamic properties of the pestivirus genus. Brief report.

The sedimentation coefficient and buoyant density of hog cholera, bovine viral diarrhea and Border disease viruses, have been compared with those of representative members of the family togaviridae. It appears that the pestivirus genus is a homogeneous group which is not only antigenically but also structurally, unrelated to the other genera of the togavirus family.


Development in vitro of cytotoxic lymphocytes against murine cytomegalovirus.

Lymphocytes cytotoxic for mouse embryo fibroblasts (MEF) infected with murine cytomegalovirus (MCMV) were produced by in vitro culture of "memory" spleen cells with UV-irradiated, MCMV-infected, MEF. Cytotoxic lymphocytes were developed from spleen cells of mice 10 to 240 days after infection with MCMV. The cytotoxic cells carried the theta and Ly 2 antigens, and were H-2 restricted in the recognition of infected target cells.


[Microbiological investigations into soil in a fertilizer trial with spruce in the fume-damaged zone of the Erzgebirge (author's transl)].

In the heavily fume-damaged spruce stands on crest sites in the eastern parts of Erzgebirge the composition of microbe population and organic matter conversion in the A0 horizon can be positively changed with a lasting effect by fertilization using CaNP. Ammonification, nitrification, urea transformation, and decomposition of cellulose are enhanced by the increased occurrence of bacteria and actinomycetes. As a result of fertilization acidity was reduced from pH (KCl) 3.0 to 3.6 as a maximum. The influence of the nutrient status is clearly manifest in the humus form (raw humus in the case of quartz porphyry, mull-resembling moder in the case of basalt), but scarcely in the chemical and microbiological properties of the Of subhorizon. No pronounced effect of the SO2 fallout on the microbiological conditions in the A0 horizon was observed. The increased soil fertility after application of Ca and P can lead to an increased and a more stable yield of wood only under certain conditions.


Effect of daily injections of ACTH on growth and on the adrenal and lymphoid tissues of two strains of immature fowls.

1. The effects of daily injections of ACTH (30 IU/kg) or physiological saline on growth and on adrenal and lymphoid tissues of Rhode Island Red (RIR) and Light Sussex (LS) chickens were compared at 1, 2 and 3 weeks of age. 2. Saline injections retarded skeletal growth in both strains during the 1st week but only affected weight gain in LS birds. 3. Injections of ACTH depressed growth rate, caused adrenal enlargement and deplenished adrenal cholesterol to the same extent in both strains. 4. ACTH caused a greater reduction in bursal size in RIR than in LS birds. Spleen size was reduced by ACTH only in RIR birds. 5. At 2 and 3 weeks the plasma concentrations of corticosterone of RIR birds were greater than those of LS birds. Plasma corticosterone concentrations were within the normal range 24 h after the last injection of ACTH in both strains. 6. Plasma concentrations of growth hormone was unaffected by ACTH treatment in RIR birds, but it was increased in LS birds after 3 weeks of treatment.


The efficacy of two commercial feline rhinotracheitis-calicivirus-panleukopenia vaccines.

The efficacy of two commercial feline vaccines was determined by challenging vaccinated and unvaccinated cats sequentially with a virulent feline calicivirus and rhinotracheitis virus. Serological responses to these viruses as well as to panleuk openia virus were also measured. Results show significant protection and satisfactory serological responses are conferred by both vaccines. One vaccine showed significant superiority in protection against feline viral rhinotracheitis.


Uridine kinase activity in human tumors.

The activity of uridine kinase, a key enzyme in the salvage pathway for pyrimidine bases and nucleosides and for the activation of the corresponding chemotherapeutic analogs, varied 10-fold in a series of human colon adenosarcomas; similar variations were observed with other tumor types. In contrast to the leukemias where only the adult isozyme appears to be present, both the adult and embryonic forms were present in the solid tumors examined. The qualitative and quantitative differences may account, in part, for differences in the innate (initial) sensitivity of the tumors to pyrimidine base and nucleoside analogs.


Nor-MDP, saponin, corynebacteria, and pertussis organisms as immunological adjuvants in experimental malaria vaccination of macaques.

Vaccination of primates against malaria using antigen derived from erythrocytic parasite stages has been most successful where Freund's complete adjuvant has been employed. Since this adjuvant is clinically unacceptable its replacement is a matter of urgency.In the present work a muramyldipeptide derivative (nor-MDP) given in mineral oil has proved to be partially effective as an adjuvant for merozoite vaccination of Macaca mulatta against Plasmodium knowlesi, and saponin has proved to be effective in similar vaccination of M. fascicularis.


[5-Bromodeoxyuridine inhibition of the transformation of Rous sarcoma virus-infected chicken cells].

The transformation of Rous-Sarcoma-Virus-infected synchronized Chicken cells is prevented when 5-bromodeoxyuridine is added for 3 hrs. at time of the peak of the first S phase following infection. Virus progeny is normally released. The inhibitory effect of BrdU is reversed by simultaneous addition of thymidine, but not by that of deoxyuridine. Hence, the inhibition of the transformation seems to result from the incorporation of the analogue into the nuclear DNA.


[Involvement of endorphins in the emotional behavior of pigs].

Social isolation and exposure to a new environment induce hyperactivity, vocalizations and ACTH release in weaned Piglets acutely submitted to this stress. These reactions are increased by pretreatment with morphine (0.5 and 1 mg/kg) and decreased by naloxone (1 mg/kg), suggesting that endogenous opiates modulate emotional behaviour in Pigs.


Reliability and reproducibility of interpretation of 99mtechnetium pyrophosphate myocardial scintigrams.

The interpretations of 156 99mtechnetium pyrophosphate myocardial scintigrams by four observers were analyzed in order to determine the reliability and reproducibility of the subjective process of reading scintigrams. The scintigrams were scored on an integral scale from 0 to 4, depending upon the degree of myocardial radionuclide accumulation, and the site and nature of uptake were specified. Exact agreement upon score was generally poor but approximate concurrence of interpretation was good (90.4 and 92.5% inter- and intra-observer agreement, respectively). There was somewhat less agreement on scintigrams with the higher scores of 3 and 4 (83.3 and 78.0%, respectively). A high level of concurrence upon the differentiation between diffuse and localized uptake, and upon the site of uptake, was found. We conclude that only approximate rather than exact agreement of individual readers' interpretations can be expected in this subjective technique, that scintigrams with higher degrees of radionuclide accumulation produce slightly greater observer disagreement, and that variability of interpretation could account for some of the diagnostic inaccuracy of 99mtechnetium pyrophosphate myocardial scintigraphy.


Metabolism of low density lipoproteins in human fibroblast culture.

The purpose of the study was to investigate the changes which low density lipoproteins (LDL) underwent during incubation with human culture fibroblasts. The results showed a predominant deposition of lipids, a low level of LDL-protein breakdown products in the cells, the altered structure of lipoproteins, absorbed on the cell surface and of lipoproteins in the incubation medium. The authors suppose intensive metabolic transformations to take place on the cell membrane. The partial lipoprotein degradation seems to realize favourable conditions for the subsequent entry of lipids into the cells and for a pinocytosis of lipoprotein particles transformed on the cell surface. The intensity of these membrane metabolic transformations is influenced by the antigenic properties of LDL, the presence of other proteins in the medium, and by the maturity and the metabolic activity of the cultured cells.


Brain opiate receptor concentrations are increased in adult spontaneously hypertensive rats.

The saturable binding of 3H-naltrexone in the brains of eight week old spontaneously hypertensive rats (SHR) is about twice that measured in corresponding normotensive WKY rats. This increase is dependent on age since in three and four week old SHR and WKY rats no difference in binding is observed. Scatchard analysis of the saturation curves for the adult animals revealed that the change in binding is due to an increase in the number of binding sites and does not reflect a difference in binding affinity. The increase in opiate receptor content of SHR rats coincides with the appearance of elevated blood pressure in these animals, and supports a concept in which an interaction between the endorphins and the endocrine system may be involved in the mechanisms controlling hypertension.


Multiplication stimulating activity regulates ornithine decarboxylase in isolated porcine granulosa cells in vitro.

The polypeptide growth factor, multiplication stimulating activity (MSA), stimulates ornithine decarboxylase (ODC) activity in isolated porcine granulosa cells maintained under chemically defined conditions in vitro. The stimulatory action of MSA is saturable, and dose-dependent (0.1-1000 ng/ml). MSA effects are additive to those of luteinizing hormone (LH), but not those of follicle stimulating hormone (FSH). Stimulation of ODC activity by MSA requires cellular protein and RNA synthesis, and appears to be mediated independently of cyclic AMP. These observations provide the first demonstration of MSA action in the mammalian ovary.


[Factor(s) controlling the secretion of adrenocorticotropin (ACTH) in peripheral plasma (author's transl)].

The factor(s) controlling the secretion of ACTH in peripheral plasma are not well known. The effects of non-extracted and extracted plasma on ACTH secretion were investigated using rat anterior pituitary cell cultures. Medium ACTH was assayed by radioimmunoassay, and the corticotropin releasing activity (CRA) was expressed as ACTH released. One hundred ul of non-extracted plasma showed significant CRA, whereas greater volumes of plasma showed reduced activity. Non-extracted plasma (250 approximately 500 microliter) rather reduced the secretion of ACTH evoked by hypothalamic extract (HE). When plasma was extracted with 0.2 N-acetic acid-acetone and divided into an acid phase and an acetone-ether phase by adding ether, the CRA was recovered in the acid phase while no significant activity was observed in the acetone-ether phase. The acid phase extract of plasma showed a positive dose-response relationship between the amount of plasma extract (50 approximately 800 microliter plasma equivalent) and ACTH release in pituitary cell cultures. The organic phase of plasma extract inhibited HE-induced release of ACTH, and this ACTH-release inhibiting activity was presumed to be corticosterone. When the acid phase extract of 20 ml plasma was applied to a Sephadex G-25 (fine) and eluted with 0.2 N acetic acid, two peaks of CRA were observed. One eluted in the region of void volume and another eluted in the retarded region where no activity was found in chromatography of HE. HE increased both ACTH and cyclic AMP release, but the plasma extract reduced cyclic AMP release. These results suggest that plasma contains both CRA and ACTH release inhibiting activity which can be extracted separately, and that plasma CRA is different from the hypothalamic corticotropin releasing factor.


Stability of IgG solutions, especially anti-D gammaglobulin, with special reference to measured proteolytic activity and degradation.

The stability of IgG solutions during the dating period can be predicted by measuring the degree of degradation after elevated storage conditions at +37 degrees C with added streptokinase. The measurement of proteolytic activity with a synthetic substrate correlates with the results obtained by SDS-disc gel electrophoresis and gel filtration. IgG molecules having Anti-D activity were observed to be essentially more sensitive to proteolytic degradation by plasmin than IgG molecules in general. It seems, therefore, that the methods which are suitable for the study of the stability of normal IgG preparations are not sufficient to predict the stability of anti-D gammaglobulin preparations.


Unidirectional gene conversion associated with two insertions in neurospora crassa mitochondrial DNA.

The mitochondrial phenotype of [poky] and other extranuclear Neurospora mutants is known to predominate over that of wild type in heteroplasmons. In the present work, we have investigated the interaction between wild-type and [poky] mtDNAs using as many as four physical markers to distinguish the two types of mtDNAs. Two insertions, one of 1200 bp in Eco RI-5 and the other 50 bp in Eco RI-9, are identified as sites of high frequency, unidirectional gene conversion leading to their spread through mtDNA populations in heteroplasmons. However, the transmission of the [poky] mutation does not appear to be correlated with the transmission of either of these insertions or of other physical markers. The possibility that other loci of nonreciprocal recombination might be responsible for the "dominance" of Neurospora extranuclear mutants is discussed.


Site-specific instability in Drosophila melanogaster: the origin of the mutation and cytogenetic evidence for site specificity.

During a study of delayed mutations, an unstable X chromosome (Uc) was detected. Spontaneous X-linked recessive lethal mutations were detected in 34 of 993 sperm sampled from 50 males carrying this chromosome. All but three of the 34 lethals originated as clusters in three of the 50 males Cytogenetic and complementation analyses revealed 14 intrachromosomal rearrangements: ten inversions, two reverse repeats, one deficiency and one transposition. Eight of the 14 rearrangements have one break in the 6F1-2 doublet and two rearrangements have a break in 6F1-5 of the X chromosome. The remaining four rearrangements have in addition to the aberrations a lethal point mutation between 6F1 and 6F5. Though each of the lethal lines was established from a single lethal-bearing female, chromosome polymorphism is evident in 17 of the 18 lines having rearrangements, with certain aberrations recurring in several lines. The lethal mutations revert frequently to the nonlethal state, and cytological evidence indicates that more than one mutational event may occur at the unstable locus of the chromosome during one generation. Two lethal lines had more than one type of chromosome rearrangement sharing a common breakpoint. These observations are consistent with the view that the instability of the Uc lines is caused by a transposable element capable of site-specific chromosome breaks and perpetual generation of mutations. The mutagenic and genetic properties of transposable elements can be related to the two-mutation theory of KNUDSON (1971) for cancer initiation.


Anatomy of the RNA and gene products of MC29 and MH2, two defective avian tumor viruses causing acute leukemia and carcinoma: evidence for a new class of transforming genes.

The RNA species of the defective avian acute leukemia virus MC29 and of the defective avian carcinoma virus MH2 and of their helper viruses were analyzed using gel electrophoresis, fingerprinting of RNase T1-resistant oligonucleotides, RNA-cDNA hybridization and in vitro translation. A28S RNA species, of 5700 nucleotides, was identified as MC29- or MH2-specific. MC29 RNA shared 4 out of about 17 and MH2 RNA at least 1 out of 16 T1-oligonucleotides with several other avain tumor virus RNAs. In addition MC29 and MH2 RNAs shared 2 oligonucleotides which were not found in any other viral RNA tested. 60% of each 28S RNA could be hybridized by DNA complementary to other avian tumor virus RNAs (group-specific) but 40% could only be hybridized by homologous cDNA (specific). Src gene-related sequences of Rous sarcoma virus were not found in MC29 or MH2 RNA. The specific and group-specific sequences of MC29, defined in terms of their T1-oligonucleotides, were located on a map of all T1-oligonucleotides of viral RNA. Specific sequences mapped between 0,4 and 0,7 map units from the 3'poly(A) end and group-specific sequences mapped between 0 and 0,4 and 0,7 and 1 map units. The MC29-specific RNA segment was represented by 6 oligonucleotides, two of which were those shared only by MC29 and MH2 RNAs. In vitro translation of MC29 RNA generated a major 120 000 dalton protein and minor 56 000 and 37 000 dalton proteins. The 120 000 dalton protein shared sequences with the proteins of the avian tumor viral gag gene, which maps at the 5' end of independently replicating viruses. Since a gag gene-related oligonucleotide was also found near the 5' end of MC29 RNA, we propose that the 120 000 MC29 protein was translated from the 5' 60% of MC29 RNA. It would then include sequences of the defective gag gene as well as MC29-specific sequences. Since both MC29 and MH2 lack the src (sarcoma) gene of Rous sarcoma virusk it is concluded that they contain a distinct class of transforming (onc) genes. We propose that the specific sequences of MC29 and MH2 represent all, or part of, their onc genes because the onc genes of MC29 and MH2 are specific and represent the only known genetic function of these viruses. If this proposal is correct, the onc genes of MC29 and MH2 would be related, because the specific RNA sequence of MC29 shares 2 of 6 oligonucleotides with MH2. It would also follow that the 120 000 dalton MC29 protein is a probable onc gene product, because it is translated from MC29-specific (and group-specific) sequences and because both MC29- and MH2-transformed cells contain specific 120 000 and 100 000 dalton proteins, respectively.


The in vitro of Rous sarcoma virus RNA and function of the viral protein during the viral replication.

The gag gene and pol gene of the Rous sarcoma virus are translated in vitro from the 35S viral RNA. The env gene cannot be translated in vitro from the 35S RNA. For the in vitro translation of the src gene. 3' end fragments of the viral RNA are used. The gag protein p15 has a proteolytic activity and specifically processes its own protein precursor pr76. The gag protein p19 suppresses the in vitro translation of the pol gene.


Effect of vitamin A on plasminogen activator synthesis by chick embryo fibroblasts.

Low concentrations of Vitamin A stimulated plasminogen activator synthesis (PA) in chick embryo fibroblasts (CEF). It caused a dose dependent and reversible increase in PA synthesis in both normal CEF and CEF infected with a temperature sensitive mutant of Rous Sarcoma virus (RSV-Ts68). Both induction and deinduction of PA could be inhibited by Actinomycin D. Vitamin A also accentuated the morphological changes associated with transformation in the Rous Sarcoma virus infected cells. The effects of Vitamin A on PA synthesis were essentially similar to those of the known tumour promoter, phorbol myristate acetate (PMA). Both Vitamin A and PMA were found to act synergistically with sarcoma gene expression as far as PA synthesis was concerned.


Chromosome abnormalities in leukemia.

The consistent occurrence of nonrandom chromosome changes in human malignancies suggests that they are not trivial epiphonomena. Whereas we do not understand their significance at present, one possible role which they may fulfill is to provide the chromosomally aberrant cells with a proliferative advantage as the result of alteration in the number or location of genes related to nucleic acid biosynthesis. The proliferative advantage provided by various chromosome aberrations is likely to differ in patients with different genetic constitutions.


Leukemia specific antigens: FOCMA and immune surveillance.

In cats, horizontally transmitted viruses cause leukemia and lymphoma under natural conditions. As with other retroviruses, feline leukemia virus (FeLV) contains products of 3 major genes; the virus core gag gene products, the polymerase, and the virus envelope glycoprotein. When cells are transformed in vitro by the related feline sarcoma virus (FeSV), an additional protein, FOCMA is expressed at the cell membrane. FOCMA, which is FeSV-coded, is transformation and/or tumor specific and expressed regardless of whether or not the cells make virus or contain virus structural antigens. Lymphoid leukemia cells also express FOCMA, both when FeLV is used to induce the disease in laboratory cats and when the tumors occur under natural conditions. FOCMA is expressed on both T and B lymphoid leukemia cells, but not expressed on non-malignant lymphoid cells, even when they are infected with FeLV. About one-third of the naturally occurring lymphoid tumors of cats lack detectable FeLV proteins and varying portions of the FeLV provirus. Despite this, they regularly express FOCMA, which is the target of an immuno-surveillance response that functions effectively under most conditions. FOCMA thus provides a useful model for antigens that might be expressed in "virus-negative" leukemias of man.


Detection of integrated type-C viral DNA fragments in two primates (human and gibbon) by the restriction enzyme blotting technique.

We have shown that 1. partial provirus integration can be a possible result of a natural infection, and may serve as a model in animal systems where a viral etiology is implicated but detection of a major fraction of the virus genome is rare; 2. All human DNA contains some sequences that hybridize specifically with genomes of SiSV-SiSAV, suggesting that viruses of this group have infected humans in the past and recombined with human cellular DNA. 3. Finally, DNA from uncultured leukocytes of two leukemic patients, one being HL23, which yielded the virus HL23V in culture, was shown to have virus specific fragments related to BaEV. Another human DNA sample revealed virus specific fragments related to SiSV(SiSAV). These fragments are probably acquired by infection.


Calcium regulation in the myometrium of pregnant sheep.

Fractionation of pregnant sheep myometrium has been carried out and the fractions obtained analysed for their marker enzyme content and Ca2+ transporting properties. A fairly pure mitochondrial fraction and a very pure plasma membrane fraction were obtained. Both fractions could accumulate Ca2+ in the presence of ATP but the mechanisms were different, mitochondrial uptake had a high capacity and was inhibited by azide. Plasma membrane uptake was of low capacity and unaffected by azide. The effect of oxalate on plasma membrane suggests that there is an outwardly directed Ca2+ pump in the myometrial cell membrane. Both mitochondria and plasma membrane may be important in the Cas2+ accumulation of relaxation in this muscle.


Isozyme studies on the association of the herpes simplex virus type 1 thymidine kinase gene with human chromosomes in somatic cell hybrids.

To investigate chromosomal site(s) of integration of the herpes simplex virus type 1 (HSV-1) thymidine kinase (TK) gene in biochemically transformed [HeLa(BU25)/KOS 8-1] cells, these human cells which had been transformed by ultraviolet light-irradiated HSV-1 were fused with TK-negative mouse LM(TK-) cells, human-mouse somatic cell hybrid clones (LH81 clones 1-20) were isolated by HATG-ouabain selection and their chromosomes and isozymes were analyzed. Electrophoretic and serological analyses showed that all 20 clones expressed type-specific HSV-1 TK. Isozyme analyses on 29 gene-enzyme systems representing 22 human chromosomes revealed that all of the HSV-1 TK-positive clones expressed human aminoacylase-1 (ACY-1) and esterase D (ESD), which have been mapped to human chromosomes 3 and 13, respectively. Other human isozymes were detected in only one to four clones or in none of the clones. Chromosome analyses showed that: (1) the hybrid clones retained only a few human chromosomes; (2) a marker chromosome, designated M7, consisting of a chromosome 17 translocated to the short arm of chromosome 3, occurred in 36 out of the 41 metaphases examined of LH81-4 clones 1 to 4 and in 31 out of the 33 metaphases examined of LH81-12 clone 10; (3) a modified M7 chromosome, (M7/m), in which the distal 2/3 of the long arm of M7 was translocated to a small acrocentric mouse chromosome, was the only human chromosome found in metaphases of LH81-13 clone 17; and (4) an intact human chromosome 13 was not present in LH81-12 clone 10 or LH81-13 clone 17 cells. Counterselection with BrdUrd resulted in the isolation of subclones lacking HSV-1 TK, human ACY-1 and ESD, and the human marker M7 chromosomes. The experiments indicate that the HSV-1 TK gene is probably associated in HeLa (BU25)/KOS 8-1 cells with marker chromosome M7, but the possibility is not excluded that the segment of human chromosome 13 which codes for ESD is involved.


Luteinizing hormone-releasing factor-like immunoreactivity in islet cells and insulomas of the human pancreas.

Luteinizing hormone-releasing factor (LRF)-like immunoreactivity was demonstrated by a three-layer immunoperoxidase method in the islet cells of human pancreas and in the neoplastic cells of insulomas. Adenocarcinomas originating in the exocrine pancreas were LRF-negative. The results demonstrate another hypothalamic peptide-like material in the gastrointestinal tract, the biological role of which remains to be clarified.


Mechanism of angiotensin I converting enzyme inhibition by SQ20,881 (less than Glu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro) in vivo. Further evidence for extrapulmonary conversion.

The mechanism by which the angiotensin I (AI) converting enzyme inhibitor SQ20,881 (less than Glu-Trp-Pro-Arg-Pro-Glu-Ile-Pro-Pro) blocks the pressor response to exogenous AI was studied in vivo in the intact anesthetized dog. When administered as a single dose 250 times that of injected AI (250 nmoles/kg) into either the pulmonary or systemic circulation, SQ20,881 produced inhibition of pulmonary conversion of exogenous AI to AII that lasted for more than 6 hours as judged by the absence of immunoreactive or labeled AII in the pulmonary venous effluent. In contrast, the pressor response to exogenous AI began to reappear within 1 hour of SQ20,881 administration. Six hours following SQ20,881, the pressor response to AI had nearly returned to normal, still in the absence of demonstrable intrapulmonary conversion and without release of detectable amounts of AII into the pulmonary venous effluent. These experiments demonstrated that AI has a pressor effect in the presence of SQ20,881 that is independent of pulmonary conversion. Studies with (Des-Asp) AII and (Des-Asp, Arg) AII showed that the delayed pressor response to AI following SQ20,881 administration could not be accounted for by circulating peptide metabolites of AI or AII. A competitive inhibitor of AII, (D-Asp, Ile) AII completely blocked the returning pressor response, suggesting that extrapulmonary generation of AII was responsible. The data strongly suggest that the systemic vascular bed taken as a whole contains large amounts of AI converting enzyme that is capable of rapid generation of AII without releasing the peptide into circulation. The extrapulmonary enzyme is more resistant to long-lasting blockade by SQ20,881 than pulmonary converting enzyme. The physiological role of extrapulmonary conversion systemic and local circulatory homeotasis remains to be assessed.


Bioactivities and the effect of dilution on various milk-borne murine mammary tumor viruses.

Infectivity titrations of milk-borne murine mammary tumor virus (MuMTV) from different sources or prepared in different ways or stored for periods of time have been compared. Titration curves were in general reproducible for MuMTVs of different sources or handled in different ways and for different methods of measurement, such as hyperplastic alveolar nodule (HAN) development, tumor development, or MuMTV antigen secretion in third-lactation milk, The curves had characteristic shapes with a low incidence of infection at low dilutions of milk, high incidences at intermediate dilutions, and low incidences at high dilutions. Infectivity incidences were unaffected by dilution over the range 10(-2) to 10(-5). The curves did not change appreciably with time of storage of milk at liquid N2 temperature for periods up to 3 years. Rate zonal fractionation of RIII milk gave zones with bioactivities which were not proportional to B-particle content. Upon dilution, the bioactivity of Zone 3, rich in B particles, and Zone 5, poor in B particles, increased, while the bioactivity of all the other zones usually decreased with dilution. The low incidence of infection at low dilutions may have been due, in part, to an immune response of the inoculated mouse. Administration of inactivated virions 4 h prior to or with MuMTV inoculations gave some evidence in support of this hypothesis but the complexity of the bioassay system for MuMTV lends uncertainty to interpretation of results.


Converting enzyme inhibition with an orally active compound in hypertensive man.

The short-term cardiovascular and endocrine effects of an orally active angiotensin converting enzyme inhibitor, SQ14,225, were evaluated in 17 subjects with drug-resistant hypertension (10 with essential and seven with renovascular hypertension). On normal dietary sodium, SQ14,225 (after 3 days at average dose of 664 mg/day) reduced mean arterial pressure (MAP) significantly (from 141 +/- 4 to 122 +/- 4 mm Hg, (SE), p less than 0.001). However, only eight of the patients achieved blood pressures within the normotensive range. Of eight patients with residual hypertension, seven exhibited further decreases in MAP (from 132 +/- 4 to 108 +/- 6 mm Hg (SE), p less than 0.001) when dietary sodium was reduced to 10 mEq/day. No rebound hypertension was noted when treatment was temporarily discontinued for 3 days in 11 patients. The reductions in blood pressure were not associated with either orthostatic hypotension or interference with baroreceptor reflexes. The values of supine plasma renin activity (PRA) were not always predictive of blood pressure responsiveness to the drug. With treatment, plasma aldosterone concentrations (PAC) decreased modestly (values from 40 +/- 9 to 22 +/- 3 ng/dl (SE), p less than 0.05). The plasma concentrations of cortisol, norepinephrine and serum potassium were left unchanged during the period of studies. The present study has not defined the exact mechanism by which SQ14,225 lowered blood pressure. Nevertheless, it indicates that this agent may be a practical therapeutic adjunct in the treatment of certain subsets of the human hypertensive population. The lack of serious interference with cardioscular and humoral homeostasis adds to its attractiveness as a therapeutic agent.


Membrane dynamic alteration associated with the tumorigenicity of polyoma-transformed and revertant hamster cells.

Steady state fluorescence polarization studies were carried out on membranes of polyoma-virus-transformed, revertants, re-revertants, and normal golden hamster cells. Four clones of revertant cells which exhibit different levels of reversion were isolated. The degree of reversion in these revertant clones was characterized in vitro by their contact inhibition behavior and in vivo by their tumorigenicity to hamsters. A good correlation between these two criteria was observed. Re-revertant cells were obtained from that revertant clone which exhibited the highest degree of reversion (similar to normal cells) and which could still produce tumors in hamsters. Re-revertants resembled the polyomavirus-transformed cells in both contact inhibition and tumorigenicity. Fluorescence polarization values of 1.6-diphenyl-1,3,5-hexatriene-labelled cells correlated well with cell tumorigenicity: normal and revertant cells which were not malignant exhibited distinctly lower fluorescence polarization values than those of the tumorigenic transformed and re-revertant cells. Intermediary revertant clones exhibited fluorescence polarization values in between those of normal and transformed cells, yet indistinguishable within experimental uncertainty.


Activation of mouse macrophages by pyran copolymer and role in augmentation of natural killer activity.

Inoculation of mice with pyran copolymer resulted in activation of natural killer (NK) cells as well as macrophages. Conditions optimal for the boosting of NK activity seemed to differ from those optimal for macrophage activation as assessed by cytostasis of tumor target cells. Peak levels of macrophage cytostatic reactivity were found at about 7 days after drug injection and were only achieved by the highest doses of pyran tested. Macrophage activation was consistently higher in the peritoneal cavity than in the spleen, regardless of route of administration, in contrast to the failure of i.v. pyran to induce high NK reactivity in peritoneal exudate cells. At 2-3 days after pyran treatment of older mice, NK augmentation reached peak levels, but only minimal macrophage activation was found. Despite these differences, macrophages played a role in regulating NK activity in pyran-treated mice. Functional macrophages appeared to be required for augmentation of NK activity by pyran, since boosting was impaired by prior in vivo inoculation of silica. Macrophages also appeared able to inhibit NK activity. In younger mice that exhibited high spontaneous levels of NK activity, pyran treatment produced a substantial reduction in NK activity to levels below those of untreated mice. This depression coincided with the time of peak levels of macrophage cytostasis. Furthermore, removal of adherent cells from the spleen cells of these pyran-treated mice resulted in levels of NK activity almost as high as those of untreated mice. The possibility that the depression of NK activity in young mice by pyran copolymer is due to suppressor cells is discussed.


Effects of 645 MeV and 9.2 GeV protons on Artemia eggs.

Developmental capacities of Artemia eggs have been studied after exposure to 645 MeV or 9.2 GeV protons. Effects of proton irradiation were studied in comparison with 60Co gamma ray irradiation, endpoints being emergence, hatching and 4-5 day old live nauplii percentages. Effectiveness of 645 MeV protons is greater than that of 9.2 GeV protons. R.b.e. values calculated for nauplius survival is 2.3 for 645 MeV protons and 1.5 for 9.2 GeV protons. These results can be taken into account in radiation hazard estimation during space flights.


Routine reversed-phase high-performance liquid chromatographic measurement of urinary vanillylmandelic acid in patients with neural crest tumors.

A rapid and simple reversed-phase high-performance liquid chromatographic procedure for the determination of vanillylmandelic acid (VMA) is described. This method was applied in the determination of the VMA content in urine from normal subjects and patients with neural crest lesions. Sample preparation is minimal and the analysis is short (20 min) and reproducible. The sensitivity of the UV detection is in the ng range. By this technique, fourteen adult control subjects were found to excrete a mean of 2.86 microgram VMA per mg creatinine, whereas twelve patients with pheochromocytoma excreted a mean of 15.7 microgram VMA per mg creatinine.


DEAE-silica gel and DEAE-controlled porous glass as ion exchangers for isolation of glycolipids.

DEAE-silica gel and DEAE-controlled porous glass have been used for the quantitative isolation of gangliosides and neutral glycosphingolipids from animal tissues and cells. A direct comparative study between DEAE-silica gel, DEAE-controlled porous glass and DEAE-Sephadex was made; the results indicated that DEAE-silica gel is preferable to the other two ion exchangers. DEAE-silica gel has also been found to be suitable for the fractionation of ganglioside mixtures.


Detection of immunoglobulin M antibodies to hepatitis A virus by enzyme-linked immunosorbent assay.

An enzyme-linked immunosorbent assay for the detection of immunoglobulin M (IgM) antibodies to hepatitis A virus is described. The test uses the principle of binding of IgM antibodies to anti-IgM-coated microtiter plates to determine whether the IgM antibodies attached have specificities for hepatitis A virus. In three patients with hepatitis type A followed up to 12 months, IgM antibodies to hepatitis A virus could be demonstrated from the onset of illness and during the following 2 to 3 months. When acute-phase sera from 48 patients with acute hepatitis were tested, IgM antibodies to hepatitis A virus could only be demonstrated in 18 patients previously classified as type A, whereas 30 patients with type B and non-A non-B hepatitis were negative. IgM antibodies to hepatitis A virus could not be demonstrated in 108 normal sera nor in 55 sera containing rheumatoid factor. These results indicate that the enzyme-linked immunosorbent assay for IgM antibodies to hepatitis A virus is useful in the serodiagnosis of acute hepatitis type A on a single serum sample taken during the acute phase of illness.


Enzyme-linked immunosorbent assay for serology of infectious bovine rhinotracheitis virus infections.

An enzyme-linked immunosorbent assay was developed to detect antibodies to infectious bovine rhinotracheitis virus in bovine sera. The test used viral antigen purified in a sucrose density gradient, and the antibody titers detected were up to 100 times higher than those seen in the serum neutralization test. When 250 sera were tested by those two tests, 146 were positive and 49 were negative in both tests. Only 1 serum was negative in the immunoassay test and positive in the neutralization test. However, 54 sera negative in the neutralization test were positive in the immunoassay test. These positive results appear to be in agreement with the increased sensitivity of the enzyme immunoassay.


Analysis of nonspecific reactions in enzyme-linked immunosorbent assay testing for human rotavirus.

Solid-phase enzyme immunoassays can be utilized to detect antigens directly in clinical specimens. However, a small number of stools which we tested for human rotavirus by enzyme-linked immunosorbent assay (ELISA) were found to have nonspecific activity in the absence of rotaviral antigen. Similar nonspecific activity was found in eight of eight sera which contained rheumatoid factor. This nonspecific activity was markedly reduced by pretreatment of the specimens with reducing agents, normal goat serum, and anti-human immunoglobulin M (IgM). Thus, it is likely that these specimens contain an IgM antibody capable of reacting nonspecifically with the other components of the assay. Although pretreatment with the mild reducing agent N-acetylcysteine markedly reduced this nonspecific activity, such treatment did not reduce the specific ELISA activity due to rotavirus. Other treatments did produce a reduction in specific activity. Thus pretreatment with N-acetylcysteine offers a practical means to increase the specificity of ELISA systems without reducing their sensitivity.


Enzyme immunoassay for detection of antibody to Epstein-Barr virus-specific early antigen.

Antibody to Epstein-Barr virus-induced early antigen could be measured in sera using enzyme-linked antibody. The sensitivity of this assay was comparable to that of the indirect immunofluorescent technique. Measurement of early antigen was accomplished on a producer cell line which was specifically treated to block late gene expression. It is now possible to measure Epstein-Barr virus-induced early antigen and viral capsid antigen antibodies by using the same cell line.